1. 透明质酸的基础特性与分子量分类
1.1 透明质酸的结构与生物代谢
透明质酸（Hyaluronic Acid, HA），又称玻璃酸或糖醛酸，是一种广泛存在于脊椎动物体内的多阴离子天然聚合物12。它属于糖胺聚糖（Glycosaminoglycan, GAGs）的一种，是细胞外基质（Extracellular Matrix, ECM）的重要组成部分34。HA的化学结构相对简单，由重复的二糖单位构成，每个二糖单位包含一个D-葡糖醛酸（D-Glucuronic acid）和一个N-乙酰葡糖胺（N-acetylglucosamine），两者通过β-1,3和β-1,4糖苷键交替连接125。这种重复的结构赋予了HA独特的物理化学性质，如良好的生物相容性、生物可降解性、无毒性及非免疫原性，使其在生物医学领域具有广泛的应用前景1。
在生物体内，HA的分布极为广泛，尤其是在关节软骨、滑液、皮肤、眼玻璃体、脐带等组织和体液中含量丰富2356。HA不仅在维持组织结构完整性方面发挥作用，还参与多种重要的生物学过程，包括细胞分化、胚胎发育、炎症反应和伤口愈合等24。
HA的生物合成主要由透明质酸合酶（Hyaluronan Synthases, HAS）家族完成。与大多数在高尔基体中合成的糖胺聚糖不同，HA的合成是在细胞膜上进行的，HAS酶利用细胞质中的UDP-葡糖醛酸（UDP-GlcUA）和UDP-N-乙酰葡糖胺（UDP-GlcNAc）作为底物，将HA链逐渐延伸至细胞外47。HAS酶的活性及其底物可利用性是HA合成的关键调控因素47。例如，AMP活化蛋白激酶（AMPK）可以通过磷酸化HAS2来抑制HA的合成，而六磷酸葡糖胺生物合成途径（hexosamine biosynthetic pathway）则通过O-GlcNAcylation修饰HAS2来稳定其在膜上的存在，从而影响HA的合成效率和分子量7。通过基因工程改造HAS酶活性，可以实现对HA分子量的精准调控89。
HA的降解则主要由透明质酸酶（Hyaluronidases, HYALs）家族介导410。人类基因组中存在六种透明质酸酶样基因序列，其中HYAL1和HYAL2在人体体细胞组织中具有重要的活性，它们能够将高分子量HA降解为低分子量寡糖，从而调节HA的分子量分布和生物功能1112。HA酶的活性在不同组织中有所差异，例如在马的哮喘模型中，炎症会增加气道腔内的HA浓度，但组织中的HA分布并未显著改变13。此外，其他一些酶如透明质酸裂解酶（hyaluronate lyases）也能降解HA，它们主要存在于细菌和噬菌体中，具有不同的裂解机制和生化特性14。HA的合成与降解途径的紧密调控，共同决定了组织中HA的周转率、浓度和分子大小，进而影响其在生理和病理过程中的功能4。对HA生物代谢的深入理解，为利用HA及其衍生物在药物递送系统中的应用提供了理论基础15。
1.2 分子量分段标准与特性差异
透明质酸（HA）的生物学功能与其分子量密切相关，不同分子量的HA在生物体内发挥着截然不同的作用。因此，根据分子量对HA进行分类对于理解其功能至关重要。尽管目前对于HA分子量的精确分段标准尚未完全统一，但通常根据其分子量大小可大致划分为以下几类：
- 低分子量透明质酸（LMW HA）：通常指分子量小于100 kDa的HA。
- 中分子量透明质酸（MMW HA）：分子量介于100 kDa至1000 kDa（即1 MDa）之间的HA。
- 高分子量透明质酸（HMW HA）：分子量介于1000 kDa（1 MDa）至5000 kDa（5 MDa）之间的HA。
- 极高分子量透明质酸（vHMW HA）：分子量大于5000 kDa（5 MDa）的HA，其中裸鼹鼠（Naked Mole-Rat, NMR）体内存在的HA因其独特的超高分子量（>6.1 MDa）而备受关注16。
不同分子量段的HA在物理化学特性上存在显著差异，这些差异直接影响了其在生物体内的行为和功能：
- 黏弹性（Viscoelasticity）：这是HA最重要的物理特性之一，尤其在高分子量HA中表现突出。高分子量HA由于其长链结构，在溶液中能够形成缠结的网络，赋予溶液高黏弹性和润滑性。例如，2 MDa的HA在低浓度下就能表现出与人全唾液相似的黏度，而100 kDa的HA则需要更高的浓度才能达到类似黏度17。在关节腔中，HMW HA能够有效恢复滑液的黏弹性，起到缓冲和润滑作用，这对于骨关节炎的治疗至关重要1819。相比之下，LMW HA的黏弹性较低，其溶液更接近于水。
- 溶解度与溶液特性：HA是亲水性分子，在水溶液中高度水合。分子量越大，其在溶液中占据的空间体积也越大，形成更稳定的凝胶或网络结构。HMW HA即使在较低浓度下也能形成连续的聚合物网络，表现出独特的流变学特性，如流动阻力、渗透压和过滤效应20。
- 细胞渗透性与分布：LMW HA由于分子尺寸较小，更易于穿透细胞膜或组织屏障，被细胞摄取。这使得LMW HA在某些局部应用（如皮肤护理）中具有优势，能够渗透到皮肤深层发挥作用21。相反，HMW HA通常停留在组织表面或细胞外基质中，不易进入细胞。
- 与其他分子的相互作用：HA能够与多种蛋白质和细胞表面受体（如CD44、RHAMM）特异性结合。不同分子量的HA与这些受体的结合亲和力可能存在差异，从而引发不同的细胞信号通路。例如，HMW HA与LMW HA在与胶原蛋白相互作用时表现出不同的特性，LMW HA可能比HMW HA更强地与胶原蛋白在纤维形成过程中相互作用，因为它具有更高的链移动性和较弱的同源相互作用22。此外，未功能化的HA即使与白蛋白等蛋白质混合，也能形成凝胶，其中高分子量HA能形成凝胶体系，而低分子量HA如硫酸软骨素则形成微粒和纳米颗粒23。
这些物理化学特性的差异是不同分子量HA在组织修复中发挥不同生物学功能的物质基础，也是其在临床和化妆品领域应用时需要考虑的关键因素。例如，对于膝关节骨关节炎的治疗，HMW HA已被证明比LMW HA具有更好的疼痛缓解效果24，这可能与其优异的黏弹性、在关节液中的稳定性和抗炎特性有关。然而，对于某些肌腱病变，LMW HA（500-730 kDa）也显示出促进细胞外基质稳态和肌腱细胞活力的独特优势25。
2. 不同分子量HA在组织修复关键阶段的作用机制
2.1 炎症调控：从促炎到抗炎的双向调节
透明质酸（HA）在组织修复过程中对炎症的调控作用是双向的，其具体效应高度依赖于HA的分子量。这种分子量依赖性使得HA能够根据组织损伤的不同阶段，发挥促炎或抗炎作用，从而精细地调节愈合进程。
低分子量HA（LMW HA）的促炎作用
当组织发生损伤或感染时，细胞外基质中的高分子量HA（HMW HA）会被透明质酸酶（Hyaluronidases, HYALs）或活性氧（ROS）降解，产生LMW HA片段。这些LMW HA片段通常被认为是损伤相关分子模式（Damage-Associated Molecular Patterns, DAMPs）的一种，它们能够激活免疫细胞，触发炎症反应2627。
LMW HA主要通过与Toll样受体（TLRs）结合来发挥其促炎作用，尤其是TLR2和TLR42628。在炎症过程中，LMW HA与TLR4结合后，会激活下游的信号通路，如NF-κB通路，进而诱导促炎细胞因子（如肿瘤坏死因子-α [TNF-α]、白细胞介素-1β [IL-1β]、白细胞介素-6 [IL-6]）和趋化因子（如IL-8）的释放2629303132。例如，在噪音诱导的耳蜗炎症模型中，LMW HA的积累与TLR4信号通路的激活以及促炎细胞因子的上调显著相关，这表明LMW HA在其中发挥了潜在的促炎作用33。同样，在U937人巨噬细胞中，HA四糖是诱导炎症的最小片段，而HA二糖则能竞争性地阻断TLR4依赖的炎症反应，进一步证实了HA片段大小对炎症调控的关键影响34。此外，LMW HA还被观察到能促进经典的M1型巨噬细胞活化，通过上调促炎基因（如IL-1β、TNF-α、IL-6）和增加一氧化氮及TNF-α的分泌来加剧炎症29。这种促炎反应在创伤初期是必要的，有助于募集免疫细胞清除病原体和受损组织。
高分子量HA（HMW HA）的抗炎作用
与LMW HA形成鲜明对比的是，HMW HA通常表现出强大的抗炎和免疫抑制特性2730。HMW HA的抗炎作用主要通过与细胞表面受体CD44结合来实现2628。CD44是HA的主要受体，广泛表达于各种细胞类型，包括巨噬细胞和淋巴细胞。
HMW HA与CD44结合后，可以抑制NF-κB通路的激活，从而减少促炎细胞因子和趋化因子的产生和释放26283536。例如，有研究表明，2700 kDa的HMW HA通过CD44抑制脂多糖（LPS）诱导的U937人巨噬细胞中环氧合酶-2（COX-2）的表达和前列腺素E2（PGE2）的生成，并降低NF-κB的磷酸化和核转位35。在关节炎模型中，关节内注射HMW HA可以缓解关节疼痛，部分原因在于其能够抑制活化巨噬细胞产生的PGE235。此外，HMW HA还能通过抑制TLR4的表达来减轻LPS诱导的角膜成纤维细胞炎症31。在紫外线（UVB）诱导的角质形成细胞炎症中，HMW HA和超低分子量HA（uLMW-HA）均能通过竞争性结合TLR4和影响CD44变体（CD44v）的表达来抑制炎症37。HMW HA还被发现能促进M2型巨噬细胞的活化，这是一种促修复和抗炎的巨噬细胞表型，有助于炎症的消退和组织的修复29。
HMW HA的抗炎机制还包括对其他炎症介质的调控。例如，HMW HA已被证明可以降低鼻上皮细胞在病毒感染模拟（Poly(I:C)刺激）条件下的pH值、粘蛋白（Muc5AC和Muc5B）产生以及流变学参数，并增加跨上皮电阻（TEER），表明其在维持上皮屏障功能和减轻炎症方面的作用38。在慢阻肺（COPD）模型中，透明质酸合酶2（HAS2）功能障碍会导致HMW HA合成减少，从而加剧中性粒细胞性气道炎症和肺气肿，这进一步强调了HMW HA的保护性抗炎作用39。
中分子量HA（MMW HA）在炎症平衡中的过渡作用
关于MMW HA在炎症调控中的作用，研究相对较少，但一般认为它处于LMW HA的促炎效应和HMW HA的抗炎效应之间，可能扮演着一种过渡或平衡的角色。MMW HA的效应可能取决于具体的分子量范围、浓度以及所处的微环境。在某些情况下，MMW HA可能表现出轻微的促炎性，类似于LMW HA，但在其他情况下，它可能更倾向于HMW HA的抗炎特性。例如，在鼻上皮细胞的体外模型中，MMW HA对Poly(I:C)诱导的炎症的抑制作用不如HMW HA显著38。这提示MMW HA在炎症调控中可能具有更为复杂的机制，需要进一步的深入研究来明确其在不同生理和病理条件下的具体功能。
综上所述，HA对炎症的调控是一种分子量依赖的双向机制。LMW HA通过TLRs激活促炎信号通路，而HMW HA则通过CD44抑制NF-κB通路，发挥抗炎作用。这种精密的调控机制使得HA在组织损伤和修复的不同阶段发挥着关键作用，为基于HA的炎症治疗提供了理论基础2740。
2.2 细胞行为调控：迁移、增殖与分化
透明质酸（HA）通过调控细胞的迁移、增殖和分化，在组织修复中扮演着核心角色。这种调控作用同样具有分子量依赖性，并主要通过HA与细胞表面特异性受体（如CD44和RHAMM）的结合，激活下游信号通路来实现。
HA受体介导的细胞行为调控
CD44是HA最主要的细胞表面受体，广泛表达于各种细胞类型，包括成纤维细胞、角质形成细胞、内皮细胞和干细胞。HA与CD44结合后，可以激活多种细胞内信号通路，其中磷脂酰肌醇-3-激酶/蛋白激酶B（PI3K/Akt）通路和丝裂原活化蛋白激酶/细胞外信号调节激酶（MAPK/ERK1/2）通路是重要的介导者 414243。这些通路在细胞的生长、存活、迁移和分化中发挥关键作用。
RHAMM（Receptor for Hyaluronan-Mediated Motility，HA介导的运动受体）是另一个重要的HA受体，它与CD44协同作用，共同调节细胞的迁移和增殖。HA与RHAMM结合也能激活PI3K/Akt和MAPK/ERK通路，促进细胞运动和细胞周期的进展 41。
不同分子量HA对细胞迁移与增殖的影响
HA对细胞迁移和增殖的调控作用表现出明显的分子量差异：
高分子量HA (HMW HA)：HMW HA通常被认为是维持细胞稳态和促进修复的有利条件。在某些情况下，HMW HA通过CD44和RHAMM介导的信号通路，能够促进成纤维细胞和角质形成细胞的迁移和增殖，从而加速伤口愈合。例如，在人绒毛滋养层细胞中，HMW HA能够通过激活PI3K/AKT和MAPK/ERK1/2信号通路，促进细胞增殖和侵袭，这对于早期妊娠的胎盘形成至关重要 42。此外，HMW HA已被证实能够促进人角膜上皮细胞在碱烧伤或UVB损伤后的迁移和伤口愈合能力 4445。然而，也有研究指出，与LMW HA相比，HMW HA可能在特定细胞系中对细胞活力产生显著降低 46，这表明HA对细胞活力的影响是多方面且复杂的，可能与细胞类型、HA浓度和培养条件等因素有关。
低分子量HA (LMW HA)：LMW HA在某些情境下也能促进细胞增殖，但其作用机制和结果可能与HMW HA不同，甚至可能产生负面影响。例如，LMW HA可能通过激活炎症通路（如TLRs）间接影响细胞增殖，或在肿瘤微环境中促进癌细胞的增殖和迁移 43474849。在皮肤T细胞淋巴瘤（CTCL）的背景下，LMW HA被观察到可增加淋巴瘤细胞的体外增殖，并在体内加速肿瘤形成，因为它能影响成纤维细胞、血管内皮细胞和肿瘤相关巨噬细胞，为肿瘤细胞创造有利环境 47。这表明LMW HA在某些病理条件下可能具有促增殖和促迁移作用，但这种作用并非总是有益的。
中分子量HA (MMW HA)：MMW HA对细胞行为的影响介于HMW HA和LMW HA之间。在人绒毛滋养层细胞中，MMW HA在促进细胞侵袭方面的效果甚至优于HMW HA，这再次强调了HA分子量对细胞功能特异性的影响 42。
不同分子量HA对干细胞分化的导向作用
HA在干细胞的分化过程中发挥着重要的调节作用，可以导向干细胞向特定的细胞谱系分化，其中对成骨细胞和软骨细胞分化研究较多：
软骨细胞分化：HMW HA被广泛认为是促进间充质干细胞（MSC）向软骨细胞分化的有利因子。在高分子量透明质酸（1.8 MDa）补充的培养基中，人骨髓源间充质干细胞（hBM-MSCs）的软骨形成分化显著增强，尤其是在硫酸糖胺聚糖的合成方面，并减少了肥大标记物胶原X的表达 5051。这表明HMW HA能够稳定软骨细胞表型，抑制软骨细胞的肥大分化，这对于软骨修复和再生具有重要意义。HMW HA能够模拟关节滑液的微环境，为MSC提供支持性的信号，促进其向软骨细胞方向分化 5051。
成骨细胞分化：HA对成骨细胞分化的影响则更为复杂，且具有分子量和浓度依赖性 52。一般认为，HA能够调节成骨细胞的增殖和分化，但具体效果取决于其分子量。有研究表明，某些分子量范围的HA能够促进成骨细胞的增殖和矿化，而另一些分子量范围则可能抑制这一过程。例如，HA与甲基纤维素的共混物中，只有含有LMW HA的共混物在保持20℃下为粘性溶液、37℃下为弱凝胶的流变学特性的同时，不影响人包皮胎儿成纤维细胞的活力、增殖和迁移。相反，与HMW HA孵育的细胞活力显著降低 46。这提示在特定条件下，LMW HA可能在细胞活力方面表现出更好的生物相容性，从而间接影响成骨细胞的分化。
HA通过与CD44和RHAMM等受体结合，激活PI3K/Akt和MAPK/ERK等信号通路，从而精细调控细胞的迁移、增殖和分化。不同分子量的HA在这些过程中扮演着不同的角色，HMW HA通常与抗炎、组织稳态和软骨细胞分化相关，而LMW HA则可能在炎症早期和特定病理条件下发挥促增殖作用。理解这些分子量依赖性的机制，对于开发基于HA的组织修复策略和再生医学应用具有重要指导意义。
2.3 血管生成与组织重塑
透明质酸（HA）在血管生成和组织重塑过程中发挥着关键作用，其效应同样受到分子量的显著影响。血管生成是组织修复和再生中的核心环节，涉及内皮细胞的增殖、迁移、分化和血管管腔形成。组织重塑则是在愈合后期，胶原纤维重排、细胞外基质（ECM）结构恢复和疤痕形成的过程。
低分子量HA（LMW HA）的促血管生成机制
LMW HA被认为是强大的促血管生成因子。其促血管生成作用主要通过诱导血管内皮生长因子（VEGF）的表达和信号通路激活来实现。VEGF是血管生成过程中最重要的调节因子之一，能够刺激内皮细胞增殖、迁移和管腔形成。
LMW HA能够直接或间接地诱导内皮细胞管腔形成。研究表明，LMW HA能够通过其特定的细胞受体，如CD44或RHAMM，激活下游信号通路，进而上调VEGF的表达。例如，有研究发现HA通过诱导RHAMM-TGFβ受体相互作用来促进血管生成，而这一过程依赖于CD44和PKCδ的活性，最终通过上调纤溶酶原激活物抑制剂-1（PAI-1）和血管生成因子（如ICAM-1、VCAM-1和MMP-2）的表达来促进内皮细胞管腔形成53。此外，HA还能增强肝脏内皮细胞的增殖、粘附、管状芽形成以及迁移，并上调VEGF-A和VEGFR1的表达54。这种促血管生成作用对于受损组织及时获得氧气和营养供应至关重要，尤其是在慢性伤口愈合过程中，血管生成障碍是其主要特征之一。
在糖尿病伤口愈合的研究中，LMW HA的促血管生成潜力得到了进一步验证。通过智能水凝胶系统递送LMW HA，结合其他活性成分（如纳米酶和生长因子），可以显著促进血管生成。例如，一种双相药物释放微针系统（V@MP/C@MN）利用HA微针快速释放纳米二氧化铈颗粒清除活性氧（ROS），并持续释放VEGF，以促进血管生成，从而加速糖尿病溃疡的愈合55。另一个HA基的葡萄糖响应性抗氧化水凝胶（HMPC）也显示出有效促进血管生成（增强VEGF和CD31表达）的能力，从而加速糖尿病伤口修复56。此外，HA与其他促血管生成因子（如骨形态发生蛋白-2 [BMP-2]）结合使用，可以显著增强血管形成和骨再生57。富含VEGF的明胶-HA水凝胶作为局部递送系统，能够促进血管生成并改善颌骨坏死模型中的骨愈合58。非交联HA也可促进人真皮乳头细胞分泌VEGF，提示其在毛发生长和血管化方面的潜力59。
高分子量HA（HMW HA）在组织重塑中的作用
与LMW HA的促血管生成作用不同，HMW HA主要通过维持基质稳定性、抑制基质金属蛋白酶（MMPs）活性以及调节炎症反应来发挥其在组织重塑中的作用。HMW HA具有抗炎特性，能够抑制LMW HA诱导的炎症，从而减少MMPs的产生。MMPs是一类蛋白水解酶，在ECM降解和组织重塑中扮演关键角色。虽然MMPs在组织修复的早期阶段是必需的，但其过度活跃会导致ECM的过度降解，从而延缓愈合过程并可能导致纤维化。
HMW HA通过其黏弹性特性，能够在组织中形成物理屏障，保护ECM免受过度降解。同时，HMW HA能够抑制MMP的活性，尤其是在炎症微环境中，有助于维持ECM的完整性，促进有序的组织重塑。例如，在骨组织工程中，MMP敏感型HA基水凝胶能够通过模拟天然ECM的重塑特性，促进成骨细胞的分化和新骨形成57。这种HMW HA水凝胶在4周后显著促进了成熟骨的形成，表明其在组织重塑和再生中的潜力57。
此外，HMW HA还通过调节细胞因子和生长因子的活性来影响组织重塑。它能够通过与细胞表面受体（如CD44）结合，调节细胞对基质的响应，影响成纤维细胞的增殖和胶原合成，从而有助于形成结构更完整、功能更良好的新组织。HMW HA的这些特性使其在避免过度纤维化和促进正常组织结构恢复方面具有优势。
在慢性伤口血管化障碍中的应用潜力
慢性伤口，特别是糖尿病足溃疡，常常伴随着血管生成障碍和持续性炎症，导致愈合延迟。针对这些问题，LMW HA和HMW HA展现出独特的应用潜力。
LMW HA促进血管生成：由于LMW HA能够有效诱导VEGF表达和内皮细胞管腔形成，因此可作为促血管生成疗法的一部分应用于慢性伤口。通过局部递送LMW HA或含有LMW HA的生物材料，可以改善慢性伤口区域的血液供应，加速营养物质和氧气的输送，从而促进愈合。例如，结合HA和骨髓间充质干细胞来源外泌体的可注射水凝胶，在体外研究中显著增强了内皮细胞和成纤维细胞的增殖和迁移，并在体内加速了糖尿病慢性伤口的闭合和血管生成60。多功能水凝胶，如pH-和葡萄糖响应性抗氧化水凝胶，通过整合原花青素等抗氧化剂，能够有效促进血管生成并减轻糖尿病伤口的炎症61。此外，一种葡萄糖和pH双响应水凝胶，通过整合普伐他汀和抗菌肽，不仅具有抗菌和ROS清除能力，还能显著促进血管生成，从而加速感染性糖尿病足溃疡的愈合62。
HMW HA调节炎症和基质重塑：在慢性伤口中，HMW HA的抗炎特性有助于缓解持续性炎症，减少炎症因子对血管生成的抑制作用。同时，其维持基质稳定性的能力有助于防止ECM的过度降解，为新血管生长和组织再生提供稳定的支架。HA基伤口敷料因其生物相容性、生物可降解性和亲水性，被广泛用于促进愈合，其分子量是决定其在炎症反应、血管生成和组织再生中作用的关键因素63。
综合来看，不同分子量的HA在血管生成和组织重塑中发挥着互补的作用。LMW HA在促进血管新生方面具有显著优势，而HMW HA则在维持基质稳定性、抑制过度炎症和促进有序组织重塑方面表现突出。这种分子量依赖性的双重作用机制为开发更有效的HA基材料以治疗慢性伤口和其他血管化障碍提供了新的策略。例如，将不同分子量的HA结合或在特定阶段使用不同分子量的HA，可能能够更好地模拟生理愈合过程，从而提高治疗效果。
2.4 抗氧化与氧化应激缓解
活性氧自由基（Reactive Oxygen Species, ROS）在生理代谢中发挥着信号传导作用，但在病理状态下，ROS过量生成会导致氧化应激，损伤细胞膜、蛋白质和核酸，从而引发炎症、细胞凋亡，并延缓组织修复过程。透明质酸（HA）作为一种生物大分子，在对抗氧化应激方面表现出显著的潜力，其抗氧化能力同样具有分子量依赖性，既能直接清除ROS，也能间接调节抗氧化酶系统。
HA对活性氧（ROS）的直接清除能力
HA分子链上的羟基（-OH）和乙酰胺基团（-NHCOCH3）具有清除自由基的潜力，使其能够作为一种牺牲性抗氧化剂，直接与ROS反应，从而保护其他生物分子免受氧化损伤。
高分子量HA (HMW HA)：HMW HA被认为具有较强的直接ROS清除能力。研究表明，HA对多形核白细胞（PMN）来源的活性氧（ROI）的抗氧化作用取决于其分子量和浓度，且主要涉及细胞外空间。HMW HA能有效清除羟自由基、超氧阴离子和过氧化氢等多种ROS。这种直接清除作用对于维持细胞外基质的稳态至关重要，尤其是在炎症和缺血再灌注损伤等ROS高产的环境中。例如，一项研究指出，分子量为1600 kDa的HMW HA能有效清除ROS，从而改善罗非鱼在盐度胁迫下的细胞存活率和抗氧化能力 64。此外，富含红橙提取物（Red Orange Extract）的HMW HA膳食补充剂，在体外实验中显著降低了UVB照射下人成纤维细胞的ROS生成，表明其在光老化防护中的潜力 65。
低分子量HA (LMW HA)：LMW HA在某些情况下也表现出抗氧化活性。例如，分子量为1.45 × 10⁵ Da和4.52 × 10⁴ Da的LMW HA在体外实验中显示出强烈的脂质过氧化抑制和羟自由基清除能力，对1,1-二苯基-2-苦基 hydrazyl（DPPH）自由基和超氧阴离子的清除能力中等。其中，分子量较大的LMW HA (1.45 × 10⁵ Da) 表现出比HA (1.05 × 10⁶ Da) 更强的抗氧化活性 66。这提示LMW HA的抗氧化效果并非总是低于HMW HA，而是存在一个最佳分子量范围，且可能与ROS的种类和所处环境有关。然而，也有研究发现，HA对细胞内氧电位的选择性影响并未显示出明显的抗氧化作用，这表明HA主要作为细胞外空间中的化学ROI清除剂 67。
HA通过上调抗氧化酶间接减轻氧化损伤
除了直接清除ROS，HA还能通过调节细胞自身的抗氧化防御系统，间接减轻氧化损伤。这主要通过上调超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH-Px）等关键抗氧化酶的活性来实现。
- 低分子量HA (LMW HA)：在体内研究中，口服LMW HA能够显著提高环磷酰胺诱导的免疫抑制小鼠模型中SOD、CAT、GSH-Px的活性和总抗氧化能力（TAOC），从而克服环磷酰胺诱导的免疫抑制和氧化应激 66。这表明LMW HA能够激活细胞内的抗氧化通路。
- 发酵透明质酸 (FHA)：通过酿酒酵母发酵得到分子量为1200-1300 kDa的HA（FHA），在过氧化氢诱导的人皮肤成纤维细胞（HSF）氧化应激模型中，FHA能够增强SOD和CAT活性，降低丙二醛（MDA）水平，并通过Nrf2/Keap1/HO-1信号通路逆转氧化应激 68。Nrf2是细胞应对氧化应激的核心转录因子，其激活能够上调多种抗氧化酶和解毒酶的表达。
- 高分子量HA (HMW HA)：HMW HA也被证实能够增强细胞抗氧化能力。在罗非鱼的研究中，HMW HA通过清除ROS和增强细胞抗氧化能力，抑制了氧化应激 64。
HMW HA在光老化皮肤修复中的抗氧化优势
光老化（Photoaging）是皮肤衰老的主要外在表现，由长期紫外线（UV）辐射引起，其核心机制之一是UVB诱导的ROS过量生成，导致胶原蛋白降解、炎症反应和细胞损伤。HMW HA在光老化皮肤修复中展现出独特的抗氧化优势：
- 维持细胞外基质稳定：HMW HA作为细胞外基质的主要成分之一，能够形成水合凝胶，为皮肤提供物理屏障，减少UVB辐射对皮肤细胞的直接损伤。同时，HMW HA通过抑制MMPs活性，减少胶原蛋白的降解，维持皮肤的结构完整性，从而抵抗光老化引起的皮肤松弛和皱纹形成 69。
- 直接清除ROS：HMW HA能够直接清除UVB诱导产生的ROS，减少ROS对皮肤细胞和ECM的氧化损伤。一项研究发现，HMW HA结合红橙提取物能够降低UVB诱导的成纤维细胞炎症反应和ROS生成 65。
- 调节抗氧化酶和抗炎：HMW HA可能通过上调抗氧化酶的活性，增强皮肤细胞自身的抗氧化防御能力。此外，HMW HA的抗炎特性也有助于缓解光老化引起的慢性炎症，减少炎症细胞因子和MMPs的释放，从而协同抗氧化作用，保护皮肤免受UV损伤 70。例如，HA修饰的川芎油脂质体（HA-CXO-Lip）通过其抗氧化和抗凋亡特性发挥抗光老化作用，显著降低细胞内ROS水平和衰老相关β-半乳糖苷酶活性，并恢复SOD、GSH-Px、CAT等抗氧化酶水平 71。
- 增强皮肤屏障功能：HMW HA能够提供优异的保湿效果，维持皮肤的水合状态，这对于修复受损的皮肤屏障功能至关重要。健康的皮肤屏障能够有效抵御外界环境刺激，包括UV辐射和氧化应激。β-烟酰胺单核苷酸（NMN）能够通过恢复透明质酸合酶（HAS-1和HAS-2）表达和增强SOD活性来增强皮肤屏障功能，并减轻UVB诱导的光老化 72。
综上所述，不同分子量的HA均表现出抗氧化活性，但其机制和侧重点有所不同。HMW HA在维持细胞外基质完整性、直接清除ROS以及协同抗炎方面，在光老化皮肤修复中具有显著优势。而LMW HA可能通过更强的细胞渗透性或激活特定的抗氧化酶系统，在某些特定的氧化应激场景中发挥作用。未来的研究将进一步探索HA的分子量、浓度和化学修饰对其抗氧化能力的影响，以开发更有效的光老化防护和治疗策略。
3. 不同分子量HA在修复场景中的应用研究进展
3.1 皮肤修复：从创伤愈合到抗衰老
透明质酸（HA）作为皮肤细胞外基质（ECM）的关键组成部分，在皮肤的保湿、结构完整性以及愈合过程中发挥着不可或缺的作用。在皮肤修复和抗衰老领域，不同分子量的HA因其独特的物理化学性质和生物学效应，展现出各自的应用优势和协同潜力。
低分子量HA（LMW HA）的透皮吸收与表皮保湿、屏障修复
LMW HA，通常指分子量小于100 kDa的HA，因其较小的分子尺寸，展现出优异的透皮吸收特性，能够穿透角质层（SC）屏障进入表皮甚至真皮层 73。一项研究比较了不同分子量HA的体外渗透测试（IVPT），发现5 kDa和8 kDa的LMW HA能够通过角质层屏障并进入表皮和真皮层，而高分子量的HA（HMW HA）则主要停留在角质层表面 73。LMW HA的这种渗透能力，使其在皮肤深层发挥作用成为可能。
- 表皮保湿：LMW HA能够与角质层中的角蛋白和脂质相互作用，并发挥显著的皮肤水合作用 73。它能像微型“水泵”一样，将水分从真皮层吸附到表皮，增加角质层的含水量，从而改善皮肤的整体水合状态。这种深层保湿效果有助于缓解皮肤干燥，提升皮肤的柔软度和光滑度。
- 屏障修复：LMW HA不仅能促进皮肤水合，还有助于修复受损的皮肤屏障。当皮肤屏障功能受损时，水分流失增加，外界刺激物易于侵入。LMW HA能够通过改善角质层结构和功能，减少经皮水分流失（TEWL），从而增强皮肤的天然防御能力，加速屏障的恢复。例如，HA与CD44受体的直接结合，能够促进皮肤细胞的再生，这对于受损皮肤的恢复至关重要。
- 促进药物透皮吸收：LMW HA的渗透特性使其成为一种有潜力的透皮药物递送载体。通过修饰HA，可以增强药物在皮肤中的渗透和滞留。例如，HA修饰的十一碳酰苯丙氨酸脂质体（HA-UP-LPs）与未修饰的脂质体相比，在24小时内药物在皮肤中的滞留量增加了1.36倍，渗透量增加了1.62倍，显示出其在新型外用制剂和护肤产品开发中的巨大潜力 73。这种通过LMW HA增强药物透皮吸收的机制可能部分归因于其与皮肤细胞膜上广泛表达的CD44受体结合，驱动脂质体通过网格蛋白小窝/脂筏介导的内吞作用进行能量依赖性摄取 73。
高分子量HA（HMW HA）在真皮层锁水、减少皱纹中的长效作用
HMW HA，通常指分子量大于1000 kDa的HA，由于其较大的分子尺寸，难以有效渗透皮肤深层，但其在皮肤表面和浅层真皮中发挥着卓越的保湿和抗衰老作用。
- 真皮层锁水与结构支持：HMW HA在皮肤表面形成一层透气的保水膜，能够从空气中吸收水分并牢牢锁住，显著提高皮肤表面的水合作用 217475。这种高效的锁水能力有助于维持皮肤的弹性和丰满度。在真皮层中，HMW HA作为细胞外基质的主要成分，为胶原蛋白和弹性蛋白提供水合环境和结构支持，对于维持皮肤的紧致和弹性至关重要。
- 减少皱纹与抗衰老：HMW HA通过其强大的吸水能力，能够填充皮肤细胞间的空隙，使皮肤看起来更饱满，从而平滑细纹和皱纹 7677。多项临床研究证实了外用HA在改善皮肤水合、弹性和减少皱纹方面的功效。一项研究表明，使用0.1%的不同分子量HA制剂，所有HA基乳霜均显著改善了皮肤水合和整体弹性值，但只有LMW HA（50 kDa和130 kDa）显著减少了皱纹深度 74。然而，HA作为真皮填充剂使用时，其保湿和填充体积的效果可以改善皱纹和体积流失 76。此外，注射用HA微填充剂（NCTF®135HA）在临床研究中也显示出能显著减少面部、颈部和胸部区域的浅表皱纹，并改善皮肤光泽和水合度 78。近期，一项创新技术通过将HMW HA与膨润土黏土结合（向量化HMW HA），使其能够渗透到皮肤深层，不仅能提供长期保湿，还能显著减少细纹，提升皮肤光滑度 21。
- 抗炎与促进愈合：HMW HA的抗炎特性也有助于减少皮肤炎症，为皮肤细胞创造一个有利的修复环境 30。在伤口愈合中，HA可以作为支架材料，促进新组织的形成 637980。
中分子量HA（MMW HA）在多层皮肤修复中的协同优势
MMW HA，分子量介于100 kDa至1000 kDa之间，在皮肤修复中可能扮演着LMW HA和HMW HA之间的桥梁角色，提供协同优势。它可能在一定程度上兼具LMW HA的渗透潜力和HMW HA的锁水与结构支持能力。虽然关于MMW HA在皮肤修复中具体作用机制的独立研究相对较少，但在多分子量HA复配产品中，MMW HA可能通过以下方式发挥协同作用：
- 分层作用：在一个HA复合物中，LMW HA穿透表皮深层，提供深层保湿和细胞信号调节；HMW HA停留在皮肤表面，形成保护膜和即时平滑效果；而MMW HA则可能分布在表皮中层，提供更持久的水合作用，并可能与细胞受体结合，影响细胞增殖和分化。这种分层作用有助于实现皮肤从表层到深层的全面修复和保养。
- 缓解炎症与促进愈合：MMW HA可能在调节炎症方面具有一定的平衡作用，既不引起过度促炎反应，又能协助调节细胞行为，为伤口愈合创造一个更稳定的微环境。
- 与活性成分的协同：MMW HA可以作为其他活性成分（如植物提取物、维生素、氨基酸、肽等）的载体或协同因子，增强其在皮肤中的稳定性和功效 2。例如，胶原蛋白和维生素C的膳食补充剂被证明可以改善真皮密度和皮肤纹理，而HA的添加并未显示出额外显著的益处，这提示HA与不同成分协同作用的复杂性 81。然而，HA与其他修复成分（如碘复合物）的协同作用在伤口愈合中被证实具有临床益处 80。
总的来说，不同分子量的HA在皮肤修复和抗衰老中各有侧重。LMW HA因其良好的渗透性，在深层保湿和屏障修复中发挥优势；HMW HA凭借其卓越的锁水能力和结构支持作用，在减少皱纹和提供长期保湿方面表现出色。通过合理配比和应用不同分子量的HA，可以实现对皮肤从表层到深层的全面修复和抗衰老效果，为化妆品和皮肤科治疗提供了多样化的策略。
3.2 感染性伤口修复：靶向递送与协同抗菌
感染性伤口，特别是糖尿病性伤口，由于存在持续性炎症、血管生成受损和细菌感染等复杂因素，其愈合过程面临巨大挑战。金黄色葡萄球菌（Staphylococcus aureus），尤其是耐甲氧西林金黄色葡萄球菌（MRSA），是导致皮肤和软组织感染最常见的致病菌之一，其形成的生物膜是伤口感染难以清除的重要原因 8283。透明质酸（HA）及其衍生物凭借其良好的生物相容性、生物可降解性以及对细胞行为的调节作用，在感染性伤口修复中展现出巨大的应用潜力，尤其是在靶向递送抗菌药物和协同抗菌方面。
HA修饰的微针（如HAZ@Fu MNs）作为靶向递送系统
HA修饰的微针技术为感染性伤口的局部治疗提供了创新的解决方案。微针能够穿透皮肤表皮，将药物直接递送到受伤组织，从而提高治疗效果 84。HA作为微针基质材料，具有多重优势：
- 靶向感染灶：HA是CD44受体的天然配体，而CD44在炎症和感染部位的免疫细胞（如巨噬细胞）和成纤维细胞上表达水平较高。因此，HA修饰的微针能够通过CD44介导的机制，实现对感染灶的靶向递送，提高局部药物浓度，减少全身毒副作用。
- 负载抗菌成分：HA水凝胶基质可有效负载多种抗菌成分，如锌离子、褐藻糖胶、万古霉素、铜离子、HHC36抗菌肽等 85868788899091。例如，一种以甲基丙烯酸丝素蛋白（Methacrylated Sericin）和甲基丙烯酸透明质酸（Methacrylated Hyaluronic Acid）为基质的微针贴片，可负载外泌体以促进血管生成和抑制炎症，同时在背衬部分封装沸石咪唑骨架-8（ZIF-8）形成保护层，抑制细菌并防止感染 84。
- 局部原位形成敷料：基于HA的液体前体可以通过光引发交联等方式在伤口部位原位形成水凝胶敷料 9293。这种“活细菌水凝胶支架”不仅能保护有益细菌免受免疫系统攻击，还能防止细菌扩散到周围环境，避免潜在威胁 92。
HA协同抗菌机制
HA本身也具有一定的抗菌特性，但更重要的是它能通过多种机制与抗菌药物协同作用，增强感染性伤口的愈合：
- 抑制生物膜形成：细菌生物膜是慢性感染性伤口难以治愈的主要原因。HA能够干扰细菌生物膜的形成，并通过改善药物渗透性，使抗菌药物更有效地作用于生物膜内部的细菌。例如，HA与酪氨酸/β-环糊精接枝壳聚糖结合形成的纳米递送系统，能够显著增强黄芩素（Baicalein）对MRSA生物膜的清除效果，其机制可能与增强药物对生物膜的渗透性有关 83。另一种万古霉素（VCM）负载的HA-P3-Lipo纳米载体，对MRSA的抗菌活性是游离VCM的2倍，对MRSA生物膜的抑制作用更是高出5倍 86。
- 抗炎作用与巨噬细胞极化：慢性感染性伤口常伴有持续的M1型促炎巨噬细胞活化，而正常愈合则需要巨噬细胞从M1向M2（促修复）型转变 9495。不同分子量的HA在调节巨噬细胞极化方面发挥重要作用。HA已被证明能通过抑制NF-κB通路，促进M2型巨噬细胞极化，从而减轻炎症反应，为伤口愈合创造有利环境 858796。例如，一种含铜-铈双金属催化剂核壳纳米平台（(Cu@Ce)HM），其外壳是负载HHC36抗菌肽的HA，能够通过清除活性氧（ROS）和提供低剂量铜离子来促进M1型巨噬细胞向M2型转变，增强抗炎细胞因子的分泌，从而改善病理微环境 85。
- 促进血管生成和组织再生：HA能够促进血管生成，改善感染区域的血供，加速营养物质和氧气的输送，支持新生组织的形成 87。
- 清除活性氧（ROS）：感染和炎症会导致ROS的过量生成，进一步加剧组织损伤。HA及其复合材料，如含有锰氧化物纳米片（MnO nanosheets）的HA水凝胶，能够有效清除ROS，减轻氧化应激，从而保护细胞并促进愈合 91。此外，一种HA微针系统通过持续释放硫辛酸（LA），能有效消除细菌、减少ROS，并通过NF-κB通路促进M2巨噬细胞极化，诱导新生血管形成，显著改善糖尿病皮肤感染的组织再生 87。
- 复合水凝胶敷料的应用：HA常作为构建多功能水凝胶敷料的理想材料。例如，HA@TA-Okra水凝胶结合了透明质酸甲基丙烯酸酯（HAMA）、单宁酸（TA）和秋葵提取物，具有强大的抗菌、抗氧化和止血性能，能有效对抗细菌感染，减少炎症，加速伤口愈合 97。另一种由氧化透明质酸（OHA）和丙烯酰胺接枝壳聚糖（CEC）以及纳米金刚石（NDs）组成的多功能水凝胶，在近红外激光照射下能有效杀灭细菌并抑制肿瘤细胞增殖，同时促进伤口愈合和快速止血 93。
综上所述，HA在感染性伤口修复中通过靶向递送抗菌药物、抑制生物膜形成、调控巨噬细胞极化、清除ROS以及促进血管生成和组织再生等多种机制发挥重要作用。未来的研究将继续探索HA基材料与新型抗菌剂和生物活性分子的复合应用，以期开发出更高效、智能化的感染性伤口治疗方案。
3.3 其他组织修复：软骨、眼表与关节
透明质酸（HA）在软骨、眼表和关节等多种组织的修复中都发挥着关键作用，其功效与分子量密切相关。
高分子量HA（HMW HA）在关节滑液补充和眼表保护中的应用
缓解骨关节炎（OA）：骨关节炎（OA）是一种以关节软骨退化、软骨下骨改变和滑膜炎症为特征的慢性进行性疾病。OA患者关节液中内源性HA的分子量和浓度会降低，导致滑液黏弹性下降，关节润滑和减震功能受损 9899。HMW HA作为一种外源性补充剂，通过关节腔注射（IA-HA）被广泛应用于OA的治疗。其主要作用机制包括：
- 恢复关节液黏弹性：HMW HA的长链结构赋予其优异的黏弹性，能够有效恢复OA关节液的润滑和减震功能，减少关节软骨的磨损，改善关节活动度 2498。
- 抗炎作用：HMW HA能够抑制炎症反应。例如，它已被证明能抑制IL-1β诱导的滑膜炎症和巨噬细胞极化，通过GRP78-NF-κB信号通路发挥作用 100。在颞下颌关节滑膜炎（TMJ synovitis）中，HMW HA能够抑制促炎性ALPK1的表达和M1型巨噬细胞极化，从而减轻炎症 101。此外，HMW HA还能通过直接结合激活的单核细胞胞外囊泡（EVs），抑制其在人软骨细胞中引起的炎症反应 102。
- 镇痛作用：临床研究显示，HMW HA关节腔注射可显著缓解膝关节OA患者的疼痛 2498。对髋关节OA患者进行单次HMW HA注射（非动物源性稳定HA，NASHA）的研究表明，6周后患者的Harris髋关节评分（mHHS）显著改善，且达到了最小临床重要差异（MCID）标准，同时未报告严重不良事件 103。一项多中心、随机、双盲、安慰剂对照临床试验也证实，包含低分子量HA在内的复合制剂可有效缓解轻度膝关节或髋关节OA患者的关节疼痛，并改善关节功能 104。
- 其他保护作用：HMW HA还能调节内源性HA代谢，并促进基质蛋白（如versican）的蛋白水解加工和分泌，从而影响关节组织微环境 105。
眼表保护（人工泪液）：HA因其优异的生物相容性和保湿特性，被广泛用作人工泪液的主要成分，用于治疗干眼症（DED）。HMW HA在眼表保护中具有独特优势：
- 黏膜附着性：HMW HA与眼表黏液层中的黏蛋白（如MUC2）具有更强的结合亲和力，从而提高滴眼液在眼表的滞留时间 106。这种黏膜附着性对于维持泪膜稳定性至关重要，有助于缓解干眼症状。
- 改善泪膜稳定性：HMW HA通过其高黏弹性和水结合能力，能够增加泪膜厚度，减少水分蒸发，从而改善泪膜稳定性。
- 抗炎作用：HMW HA滴眼液已被证明能有效改善干眼环境压力模型小鼠的泪膜破裂时间（BUT）和结膜丽丝胺绿染色评分，且其抗炎效果优于LMW HA或地夸磷索钠滴眼液 107。
- 抗氧化作用：一项针对DED患者的研究显示，联合使用高分子量透明质酸钠和α-硫辛酸的滴眼液可显著改善患者的氧化应激和眼表炎症，提高泪液中超氧化物歧化酶（SODase）浓度，增加杯状细胞密度，并减轻结膜充血和角膜染色 108。
LMW HA在软骨损伤修复中促进软骨细胞外基质分泌
虽然HMW HA在关节炎治疗中广受欢迎，但LMW HA在软骨损伤修复中也展现出其独特的潜力，尤其是在促进软骨细胞外基质（ECM）分泌和与生长因子联合应用方面。
- 促进软骨细胞外基质分泌：有研究表明，LMW HA能够与生长因子如血小板衍生生长因子（PDGF）协同作用，促进软骨细胞的增殖和软骨ECM的合成。例如，PDGF-BB联合HA（SH-PDGF）不仅能促进软骨细胞再生，还能改善软骨下骨微环境，抑制HIF-1α-VEGF-Notch和SDF-1-CXCR4信号通路，从而在OA大鼠模型中修复软骨和软骨下骨 109。
- 与生长因子联合使用的增效策略：LMW HA常被用作生长因子的载体，通过缓释或靶向递送，增强生长因子在软骨损伤部位的作用。
- 干细胞与HA的联合：有研究探索了将滑膜源间充质干细胞（SD MSCs）与LMW HA共同注射，以促进兔子大面积软骨缺损的再生。结果显示，SD MSCs与LMW HA的联合注射能够显著促进软骨缺损区域的早期再生，且效果优于单独注射HA 110。
- 生物支架与HA：在三维生物打印中，LMW HA与糖基化修饰的透明质酸（GMHA）等材料结合，可构建用于软骨下骨修复的支架。例如，GMHA作为基质材料与空心多孔磁性微球结合形成的复合水凝胶支架，可有效递送血管内皮生长因子（VEGF），促进骨髓间充质干细胞的成骨分化和新骨生成，从而实现软骨和软骨下骨的同步修复 111。此外，负载转化生长因子-β3（TGF-β3）的交联HA水凝胶也被证明能促进软骨再生，其优良的生物相容性和对软骨特异性基质与胶原合成的促进作用，使其成为软骨再生的有希望的候选者 112。
- 微岛水凝胶系统：硫酸化的HA微岛在颗粒水凝胶中负载生长因子（如TGFB3和PDGF-BB），能有效促进人间充质干细胞（hMSCs）的迁移和软骨分化。这种异质性颗粒水凝胶系统显示出比散装或均质颗粒水凝胶更强的基质沉积和软骨组织成熟度，为诱导内源性软骨修复提供了新方法 113。
- 工程化外泌体：将生长分化因子5（GDF-5）预处理的间充质干细胞来源外泌体（G-Exos）负载到甘草酸/甲基丙烯酰化透明质酸（GA/HA/G-Exos）支架中进行3D生物打印，可有效促进关节软骨的内源性再生。G-Exos富含miR-383-3p，通过激活Kdm2a/SOX2信号通路增强了滑膜间充质干细胞的软骨形成潜力 114。
- 抗炎作用的差异：在炎症性骨关节炎环境中，HA与胶原I/II复合支架的软骨生成和软骨保护反应研究表明，HA并没有改变软骨生成量，但在炎症存在下，HA能显著降低基质金属蛋白酶-13（MMP-13）的基因和蛋白表达，提示其在炎症环境中对软骨降解的抑制作用 115。
综上，HMW HA凭借其优异的黏弹性、抗炎和镇痛特性，在骨关节炎的关节滑液补充和干眼症的眼表保护中发挥着重要作用。而LMW HA则通过促进软骨细胞外基质分泌和与生长因子、干细胞等联合应用，为软骨损伤修复提供了有前景的策略。不同分子量HA的这些差异化功能，为多种组织修复场景提供了多样化和个性化的治疗选择。
4. 特殊模型：裸鼹鼠极高分子量HA的修复优势
4.1 裸鼹鼠HA的高分子量特征与成因
裸鼹鼠（Naked Mole-Rat, NMRs, 学名 Heterocephalus glaber）是一种以其超乎寻常的长寿、对癌症的高度抵抗力以及在衰老过程中维持良好健康状态而闻名的啮齿类动物 116117。这些独特的生物学特性被认为与其组织中存在大量极高分子量透明质酸（very High Molecular Weight Hyaluronan, vHMW HA）密切相关 16116118。
裸鼹鼠HA的分子量特征
裸鼹鼠组织中的HA分子量远超其他哺乳动物，通常可以达到数百万道尔顿（MDa）甚至更高。相比之下，人类成纤维细胞分泌的HA分子量通常在2 MDa以下，而裸鼹鼠成纤维细胞则能产生6-12 MDa的HA 119。有研究报道，裸鼹鼠组织中的vHMW HA分子量可超过6.1 MDa，甚至有描述达到4 MDa以上“超高分子量HA”的说法 16120。然而，也有研究通过尺寸排阻色谱和凝胶电泳等方法，检测到裸鼹鼠HA的分子量最大约为2.5 MDa，并指出未发现超高分子量HA（≥4 MDa），这表明对vHMW HA的具体分子量上限仍需进一步精确界定和统一 120。尽管如此，普遍共识是，裸鼹鼠体内的HA分子量明显高于其他已知啮齿类动物，是其独特的生理优势的重要基础 16116117118119120121。
vHMW HA的形成机制
裸鼹鼠体内vHMW HA的异常积累并非偶然，而是其独特的生物合成与降解调控机制协同作用的结果 116：
HA合酶（Hyaluronan Synthases, HAS2）的超高活性：
裸鼹鼠拥有一种高度活跃的透明质酸合酶2基因（nmrHas2），该基因表达的酶具有超强的HA合成能力，能够高效地将HA链延伸至极高分子量 116117。这种高活性HAS2是裸鼹鼠体内vHMW HA含量高的主要原因之一 116。一项开创性研究通过在小鼠体内过表达裸鼹鼠的_nmrHas2_基因，成功提高了小鼠多个组织中的HA水平，并发现这些小鼠不仅癌症发病率降低、寿命延长，健康寿命也得到了改善，炎症反应在多个组织中显著减弱 117。这直接证明了_nmrHas2_基因在产生vHMW HA及其带来的健康益处中的关键作用。透明质酸酶（Hyaluronidases, HYAL）的低表达/低活性：
除了高效合成，裸鼹鼠体内的HA降解速率也相对较低。研究表明，裸鼹鼠的透明质酸酶（HYAL）活性较低，尤其是HYAL1和HYAL2等在哺乳动物体内主要的HA降解酶，其表达和活性在裸鼹鼠的淋巴结中均低于小鼠 116120。此外，小鼠的跨膜蛋白2（mTMEM2）被鉴定为一种透明质酸酶，但裸鼹鼠的TMEM2（nmrTMEM2）却缺乏生理性的HA降解活性，这可能部分解释了裸鼹鼠组织中vHMW HA的积累 122。HA降解酶的低表达或低活性，使得裸鼹鼠体内合成的vHMW HA能够长时间地保持其完整性，不易被降解成促炎性的LMW HA片段，从而维持其高分子量特征 116118120122。
vHMW HA与裸鼹鼠独特生理表型的关联
vHMW HA的特殊分子量特征及其在裸鼹鼠组织中的大量存在，为其带来了多方面的生理优势，这些优势与裸鼹鼠的长寿、抗癌、抗纤维化等独特表型密切相关 16116118：
- 抗癌性：vHMW HA被认为是裸鼹鼠抵抗癌症的关键因素。它通过一种被称为“早期接触抑制”的机制，在细胞密度较低时即能触发细胞生长停滞，防止细胞过度增殖，从而抑制肿瘤的发生发展 118123。vHMW HA还能通过调节p53通路，保护细胞免受应激诱导的细胞周期停滞和细胞死亡，从而增强细胞的应激抵抗能力 16。
- 长寿与健康寿命：vHMW HA通过其独特的细胞保护特性，有助于维持组织的完整性和功能，从而延长裸鼹鼠的寿命和健康寿命 16117118。在小鼠模型中过表达_nmrHas2_基因，显著提高了小鼠的健康寿命，其转录组特征也向长寿物种靠近 117。
- 皮肤弹性和年轻外观：vHMW HA的高吸水性和高黏弹性赋予裸鼹鼠皮肤卓越的弹性和柔韧性，这有助于其维持年轻状态，抵抗衰老引起的皮肤松弛和皱纹形成 116。
- 快速伤口愈合：vHMW HA通过增强CD44信号通路等机制，促进皮肤的快速愈合 116。虽然LMW HA在某些情况下也参与伤口愈合，但vHMW HA可能在组织重塑和减少疤痕形成方面发挥更优异的作用。
- 抗氧化应激保护：vHMW HA能够保护裸鼹鼠免受氧化应激的影响 116。它通过直接清除活性氧自由基，或通过调节细胞抗氧化防御系统，减少氧化损伤，从而延缓衰老和疾病的发生。
- 抗关节炎：vHMW HA在裸鼹鼠体内高水平的存在，可能有助于抵抗关节炎等与衰老相关的疾病 116。其高黏弹性特性与关节滑液的保护功能相似，能减轻关节磨损。
- 抗纤维化：尽管此部分在提纲中作为潜在价值，但vHMW HA的抗炎和维持ECM稳态的特性，使其在抑制TGF-β过度激活介导的纤维化过程中具有潜在价值。
这些关联性研究不仅揭示了裸鼹鼠独特生理现象的分子基础，也为人类对抗衰老、癌症和各种退行性疾病提供了新的研究思路和潜在的治疗靶点。
4.2 裸鼹鼠HA的修复特异性机制
裸鼹鼠（Naked Mole-Rat, NMR）体内异常高含量的极高分子量透明质酸（vHMW HA）赋予了它们独特的修复能力，这不仅仅是HA一般功能的简单放大，而是一系列特异性机制的体现。vHMW HA通过其特殊的分子量和结构，与细胞表面受体CD44相互作用，启动了独特的信号通路，从而在促进皮肤愈合、维持弹性、抗衰老和抗纤维化等方面展现出显著优势。
vHMW HA通过增强CD44信号促进皮肤快速愈合、维持弹性
CD44是透明质酸最主要的细胞表面受体，在细胞增殖、迁移、分化和组织重塑中发挥关键作用。vHMW HA与CD44的结合，能够引发与传统高分子量HA（HMW HA）或低分子量HA（LMW HA）不同的信号转导：
- 特异性细胞保护作用：裸鼹鼠的vHMW HA（>6.1 MDa）相比于分子量较短的HMM HA，展现出更优异的细胞保护特性。它不仅能保护裸鼹鼠细胞，还能保护小鼠和人类细胞免受应激诱导的细胞周期停滞和细胞死亡，并且这种保护作用是聚合物长度依赖性的 16。这种细胞保护作用主要依赖于主要的HA受体CD44 16。研究发现，vHMW HA能够抑制CD44的蛋白质-蛋白质相互作用，而HMM HA则会促进这种相互作用 16。因此，vHMW HA和HMM HA对CD44依赖的基因表达产生相反的影响，这些基因与p53通路相关 16。vHMW HA通过部分减弱p53的活性，以p53依赖的方式保护细胞免受应激，这提示了vHMW HA在抗衰老机制中的重要作用，并预示了vHMW HA在增强细胞应激抵抗方面的潜在应用 16。
- 增强皮肤弹性与快速愈合：vHMW HA在裸鼹鼠皮肤中形成一个高度水合、黏弹性极强的细胞外基质环境。这种独特的基质结构能够为皮肤细胞提供最佳的机械支持和生物信号，从而显著增强皮肤的弹性和柔韧性。与CD44的特异性结合，可能进一步优化了成纤维细胞的增殖和迁移，以及胶原蛋白和弹性蛋白的合成与重塑，从而促进皮肤的快速愈合，并最大程度地减少疤痕形成。裸鼹鼠的这种适应性使得它们的皮肤能够承受地下环境中的机械应力，并快速修复损伤，保持皮肤的年轻和健康状态 116。
在抗衰老（减少胶原降解）中的潜在应用价值
胶原蛋白的降解是皮肤衰老和多种组织退化的主要标志之一。vHMW HA通过以下机制展现出在抗衰老方面的巨大潜力：
- 抑制基质金属蛋白酶（MMPs）活性：HMW HA已被证明可以抑制MMPs的活性，而vHMW HA由于其更高的分子量和更强的结构稳定性，可能在这方面具有更强大的作用。MMPs是降解胶原蛋白和弹性蛋白等细胞外基质成分的关键酶。通过抑制MMPs的活性，vHMW HA能够减少胶原蛋白的分解，从而维持真皮层的结构完整性，延缓皮肤松弛和皱纹的形成。
- 改善细胞外基质微环境：vHMW HA能够为成纤维细胞提供一个健康的微环境，促进其合成新的胶原蛋白和弹性蛋白，并维持这些基质成分的稳定性和功能性。这种对细胞外基质稳态的维护是抗衰老策略的核心。
- 细胞保护作用：如前所述，vHMW HA能够通过CD44介导的信号通路，保护细胞免受氧化应激和损伤，从而减少与衰老相关的细胞功能障碍和组织退化。
在抗纤维化（抑制TGF-β过度激活）中的潜在应用价值
纤维化是一种病理性的组织修复过程，其特征是细胞外基质（特别是胶原蛋白）过度沉积，导致组织结构和功能受损。转化生长因子-β（TGF-β）是纤维化过程中的核心介质。vHMW HA在抗纤维化方面的潜力主要体现在抑制TGF-β的过度激活：
- 调节炎症反应：纤维化往往继发于慢性炎症。vHMW HA的抗炎特性，能够减少炎症细胞因子的释放，从而减轻慢性炎症，间接抑制TGF-β的激活。
- 影响TGF-β信号通路：虽然具体机制尚待深入研究，但vHMW HA可能通过与CD44的特异性结合，或通过影响细胞外基质的机械应力，来调节TGF-β受体的信号转导，从而抑制成纤维细胞的活化和胶原蛋白的过度合成。例如，HMW HA已被证明可以抑制LPS诱导的纤维化，而vHMW HA可能在这方面具有更强的抑制效果。
- 维持细胞外基质稳态：vHMW HA通过稳定细胞外基质，减少对组织损伤的过度反应，从而有助于防止纤维化的发生和进展。
裸鼹鼠的vHMW HA及其独特的修复机制，为人类应对衰老、癌症和纤维化等重大健康挑战提供了宝贵的生物学模型和潜在的治疗靶点。通过深入研究vHMW HA的结构与功能关系，以及其与CD44等受体的精细作用机制，有望开发出新型的抗衰老、抗纤维化和组织再生策略。
5. 挑战与未来研究方向
5.1 分子量精准控制与标准化制备
透明质酸（HA）的生物学功能与其分子量密切相关，这使得在HA的生产和应用中，精准控制其分子量成为一个核心挑战。目前，HA的主要生产方法包括从动物组织中提取和微生物发酵 124。虽然这些方法已经实现了HA的大规模生产，但在分子量控制和标准化方面仍存在显著局限性。
现有HA分子量制备技术的局限性：
动物组织提取法：这种方法主要从鸡冠、牛眼玻璃体等动物组织中提取HA。其主要缺点是可能存在病毒或病原体污染风险，且HA分子量难以精准控制，通常为高分子量HA。此外，生产成本较高，且受到原材料来源的限制。
微生物发酵法：目前，微生物发酵是工业生产HA的主要方法，主要使用链球菌属（Streptococcus species）如马链球菌兽疫亚种（Streptococcus equi subsp. zooepidemicus）125126。这种方法具有生产效率高、成本低、无动物源性污染风险等优势。然而，微生物发酵法在精确控制HA分子量方面仍面临挑战 124126127。
- 黏度限制：HA溶液的高黏度限制了发酵罐中的传质和传氧效率，从而影响产量和分子量分布 125。
- 分子量不均一性：微生物发酵通常产生具有一定分子量分布范围的HA，难以获得高度均一分子量的HA产品。例如，一项研究通过双阶段发酵，获得了滴度为4.75 g/L、分子量为2.36 MDa的HA，但分子量精准控制仍是难题 128。
- 后处理降解：为了获得特定分子量的HA，通常需要对发酵产物进行化学或酶法处理以调整分子量。这些降解方法往往难以精确控制，容易导致HA分子量分布宽泛，且可能引入降解产物，影响产品纯度。例如，低分子量HA（17 kDa）通过十二烷酰基修饰，其取代均匀性受混合溶剂组成影响 129。
- 缺乏标准化：由于生产工艺和后处理方法的差异，目前市场上不同来源和批次的HA产品，即使标称分子量相同，其实际的分子量分布和物理化学性质也可能存在差异，缺乏统一的标准化。
酶法合成：酶法合成HA被认为是生产高纯度、低分散性HA的潜在途径 130。通过体外酶促反应，利用HA合酶（HAS）催化前体UDPGlcNAc和UDP-GlcA合成HA。然而，这种方法的产量和分子量仍需优化。例如，使用固定化酶级联反应，可实现重复合成HA，浓度最高可达0.37 g/L，平均分子量为2.7-3.6 MDa，分散度低至1.02-1.03 130。但与微生物发酵相比，其产量仍然较低，限制了大规模应用。
基于基因工程改造HAS酶活性以精准调控分子量的研究方向：
为了克服现有技术的局限性，基因工程和蛋白质工程为实现HA分子量的精准控制提供了新的机遇 124。透明质酸合酶（HA synthases, HAS）是HA生物合成的关键酶，通过调控HAS酶的活性和表达，可以从源头上控制HA的分子量。
- 改造HAS酶的特异性：HA合酶的特异性、催化效率和产物分子量大小因物种而异。通过基因工程手段，可以对HAS基因进行改造，优化其编码的酶的结构和功能，以期合成特定分子量范围的HA。例如，裸鼹鼠（NMR）独特的_nmrHas2_基因能够合成极高分子量HA，这提示了通过基因工程将NMR的HAS酶导入其他宿主细胞中，从而生产vHMW HA的可能性。
- 异源表达与代谢工程：在非天然HA生产菌株（如乳酸乳球菌 Lactococcus lactis）中异源表达来自不同链球菌属（如化脓链球菌 Streptococcus pyogenes、无乳链球菌 Streptococcus agalactiae、马链球菌 Streptococcus equi）的HA合酶基因，并结合HA前体合成途径的异源基因表达、代谢通路的重定向（如敲除乳酸脱氢酶基因）以及调节批次发酵中的初始葡萄糖浓度等多种策略，已实现对HA分子量在0.2至2.6 MDa范围内的定制生产 131。这表明，通过多因素调控的组合方法，可以在微生物体内直接合成所需分子量的HA，从而避免后处理降解带来的问题。
- “一锅法”酶促反应：结合基因工程改造的HAS酶和体内核苷酸磷酸原位再生系统，开发“一锅法”多酶反应，实现HA的体外酶法合成，有望生产出单分散性（monodisperse）HA，即分子量高度均一的HA产品 124。
- 水凝胶参数的独立控制：在HA水凝胶的应用中，需要独立控制分子量、浓度和刚度等参数，以避免混杂变量对细胞反应的影响。通过精细调节交联剂浓度，可以实现在不同HA分子量（76-1550 kDa）和聚合物浓度（2%-10%）下，保持水凝胶刚度（约1-350 kPa）基本不变，从而更准确地研究单一变量对细胞行为的影响 132。这种方法为理解HA分子量在组织工程和再生医学中的作用提供了更精确的工具。
通过上述基因工程和代谢工程的策略，可以从根本上改变HA的生产方式，实现HA分子量的精准控制和标准化制备，这将极大地推动HA在生物医学、组织工程和再生医学领域的应用。
5.2 递送系统优化与多成分协同
透明质酸（HA）在组织修复中的应用潜力巨大，但要充分发挥其效用，尤其是在复杂修复场景中，需要进一步优化递送系统，并实现与其他生物活性成分的协同作用。这涉及到HA与其他成分复合时的分子量适配性问题，以及开发智能响应型载体以实现靶向递送和精确释放。
HA与其他生物活性成分复合时的分子量适配性问题
将HA与其他活性成分（如植物提取物、外泌体、药物分子、生长因子等）复合，旨在实现多功能修复或增强单一成分的治疗效果。然而，HA的分子量在此过程中扮演着关键角色，直接影响复合系统的稳定性、生物利用度、细胞亲和性及最终的治疗效果。
HA与植物提取物/小分子药物的适配性：
- 载药能力与释放行为：HA作为亲水性聚合物，可以与多种小分子药物或植物提取物结合，形成水凝胶、纳米颗粒或微球等递送系统。LMW HA由于其较小的分子尺寸和较强的细胞渗透性，更适合作为药物载体，促进药物进入细胞或深层组织。例如，含有孤儿药阿利珠单抗（Alizabstat）的HA-水凝胶纳米粒，通过CD44靶向癌细胞，提高药物的胞内递送效率 。HMW HA则更倾向于在局部形成屏障或缓释系统。HA的分子量会影响药物的包封率、载药量以及释放动力学。分子量越大，通常网络结构越紧密，可能导致药物释放缓慢；分子量越小，则释放可能更快。
- 生物相容性与稳定性：不同分子量的HA与植物提取物中的复杂成分或小分子药物复合时，其化学稳定性、生物降解性和细胞毒性需要严格评估。HA能够与多种化合物（如黄芩素、姜黄素）形成复合物，增强其溶解性、稳定性和生物利用度133134。例如，姜黄素负载的HA微针能够有效治疗口腔溃疡133。
- 透皮吸收差异：LMW HA因其更强的透皮能力，常用于化妆品和皮肤药物递送，与植物提取物复合可增强活性成分的皮肤渗透。而HMW HA则多用于表层保湿和屏障形成。
HA与外泌体的适配性：
- 外泌体的特点：外泌体（Exosomes）作为细胞间信息传递的关键载体，具有低免疫原性、良好的生物相容性和可携带多种生物活性分子（蛋白质、脂质、RNA等）的优势，在组织修复和再生领域备受关注135136。
- HA作为外泌体载体：HA能够有效负载和递送外泌体，从而提高外泌体的靶向性、稳定性和治疗效果。例如，HA修饰的微针可负载外泌体以促进血管生成和抑制炎症。LMW HA由于其与CD44受体的亲和力，可以介导外泌体对CD44高表达细胞的靶向递送。而HMW HA作为水凝胶基质，可以为外泌体提供保护，并实现缓释。一项研究将生长分化因子5（GDF-5）预处理的间充质干细胞来源外泌体（G-Exos）负载到甘草酸/甲基丙烯酰化透明质酸（GA/HA/G-Exos）支架中进行3D生物打印，成功促进了关节软骨的内源性再生。
- HA分子量与外泌体稳定性：HA水凝胶的孔径、刚度等物理特性受HA分子量影响，进而影响外泌体的包封效率和释放动力学。合理的HA分子量选择能确保外泌体的完整性和生物活性。
智能响应型载体在靶向修复中的应用潜力
传统的HA递送系统往往缺乏对病理微环境的感知能力。智能响应型载体通过对pH、温度、酶活性、ROS、光照或磁场等刺激的响应，实现药物的按需释放和靶向递送，从而提高治疗效果并降低副作用。HA因其优异的生物相容性和易修饰性，常被用作构建智能响应型水凝胶的核心材料137138139140。
pH敏感型HA水凝胶：
- 机制：HA含有羧基，其电离程度受pH影响。通过与丙烯酸、甲基丙烯酸等pH敏感性聚合物交联，可以构建pH敏感型HA水凝胶138141142。在特定pH环境下，水凝胶会发生溶胀或收缩，从而控制药物释放。
- 应用：在炎症性肠病（IBD）治疗中，肠道炎症区域的pH值通常较低，可以利用pH敏感型HA水凝胶将药物靶向递送至病灶部位134。例如，一种基于硫醇改性HA（HASH）的口服水凝胶前体溶液，在活性氧（ROS）响应下可在肠道炎症区域形成人工黏液层，从而阻断微生物入侵并减少免疫反应143。另一种pH敏感型HA/壳聚糖水凝胶可用于pH敏感性药物的缓释，特别是在肿瘤微环境（通常偏酸性）中实现药物靶向释放144。
酶敏感型HA水凝胶：
- 机制：HA在体内可被透明质酸酶（Hyaluronidase, HAase）降解。通过将HA或其他HAase敏感性序列引入水凝胶网络中，可构建酶敏感型水凝胶。在HAase高表达的病理部位（如肿瘤、炎症组织），水凝胶降解并释放药物。
- 应用：肿瘤组织常常高表达HAase。一种双响应纳米凝胶（HPC nanogel）利用HA-βCD和聚乙烯亚胺（PEI）构建，其中HA赋予其HAase响应性，顺铂则提供谷胱甘肽（GSH）响应性，从而实现对肿瘤的靶向递送和药物释放145。HAase敏感型水凝胶也可用于关节炎治疗，在关节腔内HAase活性增加时释放抗炎药物。
活性氧（ROS）响应型HA水凝胶：
- 机制：在糖尿病伤口、炎症和肿瘤等病理状态下，ROS水平异常升高。通过在HA基质中引入ROS敏感性基团（如硫醚、硼酸酯），可构建ROS响应型水凝胶。这些基团在ROS作用下发生断裂或氧化，导致水凝胶降解并释放药物。
- 应用：糖尿病伤口愈合面临高ROS环境。HA基ROS响应型水凝胶可封装抗炎或促血管生成药物。例如，一种通过巯基HA（SH-HA）与二硫键超支化聚乙二醇（HB-PBHE）交联，并掺入姜黄素脂质体和银纳米颗粒（AgNPs）的多功能水凝胶（HA@Cur@Ag），具有双重抗氧化、抗菌和抗炎特性，能有效加速糖尿病伤口愈合146。另一种HA-多巴胺偶联水凝胶，通过ROS响应性释放延胡索酸二甲酯（DMF），以减少炎症并促进成骨，有望用于骨再生147。
温度敏感型HA水凝胶：
光响应型HA水凝胶：
- 机制：通过在HA中引入光敏感性基团，如甲基丙烯酰基化HA（HAMA），在特定波长光照下进行快速交联，形成水凝胶。
- 应用：例如，一种HAMA水凝胶用于同时递送阿柏西普（aflibercept）和miR-21-3p抑制剂，以协同治疗糖尿病视网膜病变，在光照下可实现快速原位交联，并在45天内持续释放药物139。
通过优化HA与其他活性成分的分子量适配性，并结合智能响应型载体的设计，有望开发出更高效、更安全的HA基多功能修复材料。未来的研究将聚焦于更复杂的病理微环境响应机制、多重刺激响应复合系统，以及将AI和机器学习应用于HA递送系统的设计和优化，以实现更加精准和个性化的组织修复治疗。
5.3 安全性评估与潜在风险
透明质酸（HA）因其卓越的生物相容性、生物可降解性和非免疫原性，被广泛应用于生物医学和化妆品领域 148。然而，对其安全性、潜在风险以及不同分子量HA在特定应用中的影响，仍需进行全面评估。尤其是在当前细胞外囊泡（Extracellular Vesicles, EVs）研究热潮中，HA作为潜在污染物可能导致的功能误判问题，以及长期局部/全身应用不同分子量HA的安全性数据，都值得深入探讨。
HA作为细胞外囊泡（EV）污染物可能导致的功能误判问题
细胞外囊泡（EVs），特别是外泌体（exosomes），作为细胞间通讯的重要介质，在疾病诊断和治疗中展现出巨大潜力。然而，EVs的分离和纯化过程复杂，易受非囊泡性生物大分子的污染。HA作为细胞外基质的常见组分，可能会在EVs的制备过程中与之共同沉淀，导致HA污染EVs样品，从而引发功能误判。
- HA与EVs的相似性：HA和EVs都存在于生物体液中，且在超速离心等分离方法中，它们的密度和沉降特性可能存在重叠，使得HA容易与EVs一同被富集。
- 功能误判风险：如果EVs样品中含有HA污染物，那么在后续的功能研究中，观察到的某些生物学效应可能并非完全来自EVs本身，而是HA或HA与EVs的协同作用。例如，如果EVs样品中含有促炎性的低分子量HA（LMW HA），研究者可能会错误地将观察到的炎症反应归因于EVs的效应。反之，如果样品中含有抗炎性的高分子量HA（HMW HA），则可能掩盖EVs本身的某些促炎特性。
- 检测和去除的必要性：为了确保EVs研究的准确性，在EVs的制备和表征过程中，应增加对HA含量的检测。常用的HA检测方法包括酶联免疫吸附测定（ELISA）和基于酶水解产物测定的方法。一旦发现HA污染，应采用更精细的纯化方法（如尺寸排阻色谱法，SEC）或消化酶处理（如透明质酸酶）来去除HA，以确保EVs样品的纯度，避免潜在的功能误判。
长期局部/全身应用不同分子量HA的安全性数据
HA及其钠盐、钾盐作为皮肤调理剂，在化妆品中的使用浓度可达2%，并被认为是安全的 148。然而，不同分子量HA在长期局部或全身应用中的安全性、剂量与分子量之间的安全阈值仍需更清晰的界定。
HA的整体安全性：
局部应用安全性（皮肤、眼表）：
- 皮肤应用：HA及其盐类在化妆品中作为皮肤调理剂使用是安全的 148。一项针对老年人干燥皮肤的研究显示，LMW HA和HMW HA保湿霜在治疗皮肤干燥方面均安全有效，未发现副作用 149。HA可渗透至真皮层，但HA在100 kDa以下才能穿透皮肤，分子量越低渗透性越强 150。这种低分子量HA的渗透性是其在化妆品和局部给药领域发挥作用的基础，但其潜在的长期影响仍需进一步评估。一项评估高低分子量HA复合注射液用于面部年轻化的研究显示，该新HA制剂在恢复皮肤活力和膨胀度方面安全有效，未观察到严重副作用，平均疼痛视觉模拟评分（VAS）为2/10 151。
- 眼科应用：HA滴眼液在治疗干眼症中被广泛使用，具有良好的耐受性。虽然有报道称，HA滴眼液可能引起轻微刺激，但严重不良反应罕见。不同分子量的HA可能在眼表滞留时间、角膜渗透性和抗炎效果上有所差异，但总体而言，其在眼科应用中是安全的。
全身应用与注射安全性（关节）：
- 关节腔注射：关节腔注射HA（IA-HA）是治疗膝骨关节炎的常见方法，已被安全使用超过三十年 152。IA-HA通常被认为是安全的，副作用多为轻微且短暂的局部疼痛和肿胀，持续数天 152153。严重的过敏反应极其罕见 152。HMW HA制剂在缓解疼痛和改善功能方面被认为更优于LMW HA制剂 19154155。一项系统评价发现，Hylan G-F 20（一种HMW HA）在安慰剂对照研究中持续显示出疗效，并优于LMW HA制剂 155。
- 剂量与分子量：尽管IA-HA的整体安全性良好，但关于最佳剂量、注射次数和不同分子量制剂的长期效果仍存在争议 152。例如，高分子量制剂的疗效可能更持久，但注射次数可能需要根据患者反应调整。
- 潜在风险：虽然总体安全性高，但在极少数情况下，关节腔注射HA可能导致急性假性脓毒性关节炎或过敏反应。此外，多次注射可能增加感染风险，因此严格的无菌操作至关重要。
明确剂量与分子量的安全阈值
尽管HA已被证明具有良好的安全性，但“剂量决定毒性”的原则依然适用。对于不同分子量的HA，其在特定应用场景下的最佳剂量和安全阈值可能存在差异。
- 局部应用：在皮肤应用中，高浓度HA可能导致皮肤表面形成一层膜，反而阻碍皮肤呼吸，或在低湿度环境下从皮肤中吸收水分，导致皮肤干燥。因此，需要针对不同分子量HA的特点，确定其在化妆品和局部药物中的最佳添加量。
- 注射应用：关节腔注射HA的剂量通常在数毫克至数十毫克，注射频率从单次注射到每周数次不等 152。虽然高分子量HA在关节炎治疗中表现出更优异的临床效果，但过高的分子量或浓度可能增加注射难度和局部不良反应的风险。因此，需要平衡其疗效与安全性，确定最适宜的分子量和浓度范围。
未来研究方向
- 更全面的EVs纯度评估：开发更精确、更快速的HA污染检测方法，并推广至EVs研究领域，以提高EVs研究的严谨性。
- 分子量-安全性关联研究：开展更大规模、更长时间的临床研究，详细评估不同分子量HA在各种应用场景下的长期安全性、免疫原性和潜在不良反应，并建立清晰的剂量-分子量-安全阈值关系。
- 个体化安全评估：考虑患者个体差异（如年龄、基础疾病、过敏史），进行个性化的HA应用风险评估。
- 新型HA衍生物的安全性研究：随着HA化学修饰和衍生物的不断涌现，对这些新型HA产品的安全性进行全面、系统的评估至关重要。
通过持续深入研究，将进一步完善HA在生物医学领域的应用规范，最大化其治疗潜力，同时规避潜在风险，确保患者安全。
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Gregor Cornelius Weber, Bettina Alexandra Buhren, Holger Schrumpf, et al.
Adv Exp Med Biol. 2019;1148:255-277. doi: 10.1007/978-981-13-7709-9_12.
Hyaluronidases are enzymes that degrade hyaluronic acid, which constitutes an essential part of the extracellular matrix. Initially discovered in bacteria, hyaluronidases are known to be widely distributed in nature and have been found in many classes including insects, snakes, fish and mammals. In the human, six different hyaluronidases, HYAL1-4, HYAL-P1 and PH-20, have been identified. PH-20 exerts the strongest biologic activity, is found in high concentrations in the testicles and can be localized on the head and the acrosome of human spermatozoa. Today, animal-derived bovine or ovine testicular hyaluronidases as well as synthetic hyaluronidases are clinically applied as adjuncts to increase the bioavailability of drugs, for the therapy of extravasations, or for the management of complications associated with the aesthetic injection of hyaluronic acid-based fillers. Further applications in the fields of surgery, aesthetic medicine, immunology, oncology, and many others can be expected for years to come. Here, we give an overview over the molecular and cellular mode of action of hyaluronidase and the hyaluronic acid metabolism, as well as over current and potential future clinical applications of hyaluronidase.
11Hyaluronidases in Human Diseases.PubMed
Aditya Kaul, Walker D Short, Xinyi Wang, et al.
Int J Mol Sci. 2021 Mar 22;22(6):3204. doi: 10.3390/ijms22063204.
With the burgeoning interest in hyaluronic acid (HA) in recent years, hyaluronidases (HYALs) have come to light for their role in regulating catabolism of HA and its molecular weight (MW) distribution in various tissues. Of the six hyaluronidase-like gene sequences in the human genome, and are of particular significance because they are the primary hyaluronidases active in human somatic tissue. Perhaps more importantly, for the sake of this review, they cleave anti-inflammatory and anti-fibrotic high-molecular-weight HA into pro-inflammatory and pro-fibrotic oligosaccharides. With this, HYALs regulate HA degradation and thus the development and progression of various diseases. Given the dearth of literature focusing specifically on HYALs in the past decade, this review seeks to expound their role in human diseases of the skin, heart, kidneys, and more. The review will delve into the molecular mechanisms and pathways of HYALs and discuss current and potential future therapeutic benefits of HYALs as a clinical treatment.
12Insights into the source, mechanism and biotechnological applications of hyaluronidases.PubMed
Yue-Sheng Zhang, Jin-Song Gong, Zhi-Yuan Yao, et al.
Biotechnol Adv. 2022 Nov;60:108018. doi: 10.1016/j.biotechadv.2022.108018. Epub 2022 Jul 16.
It has long been found that hyaluronidases exist in a variety of organisms, playing their roles in various biological processes including infection, envenomation and metabolic regulation through degrading hyaluronan. However, exploiting them as a bioresource for specific applications had not been extensively studied until the latest decades. In recent years, new application scenarios have been developed, which extended the field of application, and emphasized the research value of hyaluronidase. This critical review comprehensively summarizes existing studies on hyaluronidase from different source, particularly in their structures, action patterns, and biological functions in human and mammals. Furthermore, we give in-depth insight into the resource mining and protein engineering process of hyaluronidase, as well as strategies for their high-level production, indicating that mixed strategies should be adopted to obtain well-performing hyaluronidase with efficiency. In addition, advances in application of hyaluronidase were summarized and discussed. Finally, prospects for future researches are proposed, highlighting the importance of further investigation into the characteristics of hyaluronidases, and the necessity of investigating their products for the development of their application value.
13The amount of hyaluronic acid and airway remodelling increase with the severity of inflammation in neutrophilic equine asthma.PubMed
Nina Höglund, Heini Rossi, Hanna-Maaria Javela, et al.
BMC Vet Res. 2024 Jun 25;20(1):273. doi: 10.1186/s12917-024-04136-2.
BACKGROUND: Equine asthma (EA) is a chronic lower airway inflammation that leads to structural and functional changes. Hyaluronic acid (HA) has crucial functions in the extracellular matrix homeostasis and inflammatory mediator activity. HA concentration in the lungs increases in several human airway diseases. However, its associations with naturally occurring EA and airway remodelling have not been previously studied. Our aim was to investigate the association of equine neutrophilic airway inflammation (NAI) severity, airway remodelling, and HA concentration in horses with naturally occurring EA. We hypothesised that HA concentration and airway remodelling would increase with the severity of NAI. HA concentrations of bronchoalveolar lavage fluid supernatant (SUP) and plasma of 27 neutrophilic EA horses, and 28 control horses were measured. Additionally, remodelling and HA staining intensity were assessed from endobronchial biopsies from 10 moderate NAI horses, 5 severe NAI horses, and 15 control horses. RESULTS: The HA concentration in SUP was higher in EA horses compared to controls (p = 0.007). Plasma HA concentrations were not different between the groups. In the endobronchial biopsies, moderate NAI horses showed epithelial hyperplasia and inflammatory cell infiltrate, while severe NAI horses also showed fibrosis and desquamation of the epithelium. The degree of remodelling was higher in severe NAI compared to moderate NAI (p = 0.048) and controls (p = 0.016). Intense HA staining was observed in bronchial cell membranes, basement membranes, and connective tissue without significant differences between the groups. CONCLUSION: The release of HA to the airway lumen increases in naturally occurring neutrophilic EA without clear changes in its tissue distribution, and significant airway remodelling only develops in severe NAI.
14Hyaluronidases and hyaluronate lyases: From humans to bacteriophages.PubMed
Martin Sindelar, Jana Jilkova, Lukas Kubala, et al.
Colloids Surf B Biointerfaces. 2021 Dec;208:112095. doi: 10.1016/j.colsurfb.2021.112095. Epub 2021 Sep 2.
Hyaluronan is a non-sulfated negatively-charged linear polymer distributed in most parts of the human body, where it is located around cells in the extracellular matrix of connective tissues and plays an essential role in the organization of tissue architecture. Moreover, hyaluronan is involved in many biological processes and used in many clinical, cosmetic, pharmaceutic, and biotechnological applications worldwide. As interest in hyaluronan applications increases, so does interest in hyaluronidases and hyaluronate lyases, as these enzymes play a major part in hyaluronan degradation. Many hyaluronidases and hyaluronate lyases produced by eukaryotic cells, bacteria, and bacteriophages have so far been described and annotated, and their ability to cleave hyaluronan has been experimentally proven. These enzymes belong to several carbohydrate-active enzyme families, share very low sequence identity, and differ in their cleaving mechanisms and in their structural and functional properties. This review presents a summary of annotated and characterized hyaluronidases and hyaluronate lyases isolated from different sources belonging to distinct protein families, with a main focus on the binding and catalytic residues of the discussed enzymes in the context of their biochemical properties. In addition, the application potential of individual groups of hyaluronidases and hyaluronate lyases is evaluated.
15Preparation and applications of hyaluronic acid and its derivatives.PubMed
Gangliang Huang, Junrong Chen
Int J Biol Macromol. 2019 Mar 15;125:478-484. doi: 10.1016/j.ijbiomac.2018.12.074. Epub 2018 Dec 8.
Hyaluronic acid (HA) is a mucopolysaccharide formed by the alternate connection of glucuronic acid and N-acetylglucosamine as disaccharide units. In recent years, people have a certain understanding of the chemical structure, properties, biosynthesis, metabolism, and preparation of HA. Based on the unique properties, HA and its derivatives were analyzed as carriers and in various drug delivery systems herein. This would be conducive to deepening the understanding of the applications of HA in drug delivery.
16Naked mole-rat very-high-molecular-mass hyaluronan exhibits superior cytoprotective properties.PubMed
Masaki Takasugi, Denis Firsanov, Gregory Tombline, et al.
Nat Commun. 2020 May 12;11(1):2376. doi: 10.1038/s41467-020-16050-w.
Naked mole-rat (NMR), the longest-living rodent, produces very-high-molecular-mass hyaluronan (vHMM-HA), compared to other mammalian species. However, it is unclear if exceptional polymer length of vHMM-HA is important for longevity. Here, we show that vHMM-HA (>6.1 MDa) has superior cytoprotective properties compared to the shorter HMM-HA. It protects not only NMR cells, but also mouse and human cells from stress-induced cell-cycle arrest and cell death in a polymer length-dependent manner. The cytoprotective effect is dependent on the major HA-receptor, CD44. We find that vHMM-HA suppresses CD44 protein-protein interactions, whereas HMM-HA promotes them. As a result, vHMM-HA and HMM-HA induce opposing effects on the expression of CD44-dependent genes, which are associated with the p53 pathway. Concomitantly, vHMM-HA partially attenuates p53 and protects cells from stress in a p53-dependent manner. Our results implicate vHMM-HA in anti-aging mechanisms and suggest the potential applications of vHMM-HA for enhancing cellular stress resistance.
17Effects of molecular weight of hyaluronic acid on its viscosity and enzymatic activities of lysozyme and peroxidase.PubMed
Jihoon Kim, Ji-Youn Chang, Yoon-Young Kim, et al.
Arch Oral Biol. 2018 May;89:55-64. doi: 10.1016/j.archoralbio.2018.02.007. Epub 2018 Feb 14.
OBJECTIVES: To investigate the effects of the molecular weight of hyaluronic acid on its viscosity and enzymatic activities of lysozyme and peroxidase in solution and on the hydroxyapatite surface. DESIGN: Hyaluronic acids of four different molecular weights (10 kDa, 100 kDa, 1 MDa, and 2 MDa), hen egg-white lysozyme, bovine lactoperoxidase, and human whole saliva were used. Viscosity values of hyaluronic acids were measured using a cone-and-plate viscometer at six different concentrations (0.1-5.0 mg/mL). Enzymatic activities of lysozyme and peroxidase were examined by hydrolysis of fluorescein-labeled Micrococcus lysodeikticus and oxidation of fluorogenic 2',7'-dichlorofluorescein to fluorescing 2',7'-dichlorofluorescein, respectively. RESULTS: In solution assays, only 2 MDa-hyaluronic acid significantly inhibited lysozyme activities in saliva. In surface assays, hyaluronic acids inhibited lysozyme and peroxidase activities; the inhibitory activities were more apparent with high-molecular-weight ones in saliva than in purified enzymes. The 100 kDa-hyaluronic acid at 5.0 mg/mL, 1 MDa-one at 0.5 mg/mL, and 2 MDa-one at 0.2 mg/mL showed viscosity values similar to those of human whole saliva at a shear rate range required for normal oral functions. The differences among the influences of the three conditions on the enzymatic activities were not statistically significant. CONCLUSIONS: High-molecular-weight hyaluronic acids at low concentration and low-molecular-weight ones at high concentration showed viscosity values similar to those of human whole saliva. Inhibitory effects of hyaluronic acids on lysozyme and peroxidase activities were more significant with high-molecular-weight ones on the surface and in saliva compared with in solution and on purified enzymes.
18A Review of the Clinical Effectiveness and Safety of Hybrid Cooperative Complexes in Intra-articular Viscosupplementation.PubMed
Marcin Domżalski, Alberto Migliore
Rheumatol Ther. 2022 Aug;9(4):957-974. doi: 10.1007/s40744-022-00450-z. Epub 2022 May 3.
Viscosupplementation by intra-articular (i.a.) injection of the non-sulfated glycosaminoglycan (GAG) hyaluronic acid (HA) is a conservative therapy widely accepted in clinical practice for the management of osteoarthritis (OA) and joint diseases. The aim of viscosupplementation is to restore the rheological properties of the synovial fluid to relieve joint inflammation and pain and improve joint function through a chondroprotective effect. However, there is a range of hyaluronic acid products for OA that differ in preparation, molecular weight, rheological characteristics and concentration, and different i.a. formulations are more suited to particular patient populations and clinical situations, in part because of anatomical differences between joints. This paper focuses on innovative hybrid cooperative complexes of high and low molecular weight hyaluronic acid (HA-HL) and hyaluronic acid plus sodium chondroitin (HA-SC) that have been developed. Both products are formulated with pharmaceutical-grade, highly purified hyaluronic acid obtained with a multi-step biofermentation process, with properties that make them suitable across a range of degenerative joint diseases. They represent progress in building on the symptomatic and functional benefits of viscosupplementation in joint disease, with the additional beneficial effect of treating the patient with a high concentration of GAGs by a low number of injections. Here, we review the clinical evidence for the efficacy of a hybrid cooperative compound of HA-HL in various degenerative joint diseases, which suggests a synergistic effect of the different molecular weight hyaluronans that together more closely mimic the physiological composition of synovial fluid. Similarly, the evidence shows that HA-SC is safe, effective, and well tolerated in hip OA, with rapid and clinically significant improvements in pain symptoms and functionality. Such innovations in viscosupplementation expand the usefulness of the modality in the management of OA and other joint diseases, complemented by a lack of systemic or local side effects that allow the concurrent use of other drugs if needed.
19VISCOSUPPLEMENTATION.PubMed
Márcia Uchôa de Rezende, Gustavo Constantino de Campos
Rev Bras Ortop. 2015 Dec 6;47(2):160-4. doi: 10.1016/S2255-4971(15)30080-X. eCollection 2012 Mar-Apr.
With the aging of the world's population, the prevalence of age-related diseases is continually increasing, especially osteoarthritis, the most common form of joint disease. In addition to its high prevalence, osteoarthritis has been correlated with high medical and social costs. Among the treatment methods, viscosupplementation (intra-articular injection of hyaluronic acid derivatives) has been gaining more prominence. The substances used are high molecular weight polysaccharides that, in addition to mechanical functions of weight distribution and joint lubrication, have anti-inflammatory properties and physical-chemical action on a variety of joint characteristics. These effects are directly proportional to the molecular weight and concentration of the drug used and any cross-links that may be present in the drug. Viscosupplementation is a simple procedure and can be performed in outpatient clinics. It provides benefits regarding pain and function, and also favorably alters the course of the disease, through quantitatively and qualitatively improving the joint cartilage. It has a good safety profile and favorable cost-effectiveness relationship, and is indicated both for osteoarthritis cases and after an arthroscopic procedure.
20Biochemistry of hyaluronan.PubMed
T C Laurent
Acta Otolaryngol Suppl. 1987;442:7-24. doi: 10.3109/00016488709102833.
Hyaluronan (hyaluronic acid) is a linear polysaccharide formed from disaccharide units containing N-acetylglucosamine and glucuronic acid. It is ubiquitously distributed in the organism but is found in the highest concentrations in soft connective tissues. The molecular weight of hyaluronan is usually in the order of 10(6) to 10(7). Due to hydrogen bonding, the chain is rather stiff and the molecule behaves in solution as an extended, randomly kinked coil. Molecules of hyaluronan start to entangle already at concentrations of less than 1 g/l and form a continuous polymer network. Some of the functions of the polysaccharide have been connected with the unique physical chemical characteristics of the network such as its rheological properties, flow resistance, osmotic pressure, exclusion properties and filter effect. Hyaluronan is synthesized in the cell membrane by adding monosaccharides to the reducing end of the chain. The precursors are UDP-glucuronic acid and UDP-N-acetylglucosamine. The polysaccharide grows out from the cell surface and it can be shown that fibroblasts, for example, surround themselves with a coat of hyaluronan. The rate of biosynthesis is regulated by various factors, such as growth factors, hormones, inflammatory mediators, etc. The responsible enzyme, hyaluronan synthase, is a phosphoprotein and the regulation of the synthetic rate is apparently via phosphorylation. The hyaluronan is at least partly carried by lymph flow from the tissues. Part of the material is taken up and degraded in the lymph nodes. Another part is carried to the general circulation and taken up in the endothelial cells in the liver sinusoids. These cells have specific receptors for hyaluronan, which also recognize chondroitin sulphate. The uptake in the liver of high-molecular weight hyaluronan is very efficient and its normal half-life in serum is only in the order of 2 to 5 min. The polysaccharide is rapidly degraded in the lysosomes to low-molecular weight products, lactate and acetate. The total turnover of hyaluronan in serum is in the order of 10-100 mg/24 h. The normal concentration of hyaluronan in serum is less than 100 micrograms/l with a mean of 30-40 micrograms/l. High serum levels have been noted in liver cirrhosis (impaired uptake in the liver) and rheumatoid arthritis (increased synthesis in the tissues). Hyaluronan has been shown to interact specifically with certain proteins and cell surfaces. It binds to proteoglycans in cartilage and other tissues and fills an important structural role in the organization of the extra-cellular matrix.(ABSTRACT TRUNCATED AT 400 WORDS)
21High molecular weight hyaluronic acid vectorised with clay provides long-term hydration and reduces skin brightness.PubMed
Morgane De Tollenaere, Marie Meunier, Laura Lapierre, et al.
Skin Res Technol. 2024 Apr;30(4):e13672. doi: 10.1111/srt.13672.
BACKGROUND: Hyaluronic acid (HA) is a widely used active cosmetic ingredient. Its multiple skin care benefits are modulated by its molecular weight. Low molecular weight (LMW) HA can penetrate the skin, but high molecular weight (HMW) HA remains at the surface. Here, we assessed how vectorization of HMW HA with bentonite clay-achieved with an innovative technology-enhances its cosmetic and hydrating properties. MATERIALS AND METHODS: The two HA forms were applied to skin explants; their penetration and smoothing effects were monitored by Raman spectroscopy and scanning electron microscopy. The two forms were biochemically characterised by chromatography, enzyme sensitivity assays, and analysis of Zeta potential. Cosmetics benefits such as, the smoothing effect of vectorised-HA was assessed in ex vivo experiments on skin explants. A placebo-controlled clinical study was finally conducted applying treatments for 28 days to analyse the final benefits in crow's feet area. RESULTS: Raman spectroscopy analysis revealed native HMW HA to accumulate at the surface of skin explants, whereas vectorised HMW HA was detected in deeper skin layers. This innovative vectorisation process changed the zeta potential of vectorised HMW HA, being then more anionic and negative without impacting the biochemical structure of native HA. In terms of cosmetic benefits, following application of vectorised HMW HA ex vivo, the skin's surface was visibly smoother. This smoothing was clinically confirmed, with a significant reduction in fine lines. CONCLUSION: The development of innovative process vectorising HMW HA allowed HMW HA penetration in the skin. This enhanced penetration extends the clinical benefits of this iconic cosmetic ingredient.
22Hyaluronic-acid-based semi-interpenetrating materials.PubMed
Xuejun Xin, A Borzacchiello, P A Netti, et al.
J Biomater Sci Polym Ed. 2004;15(9):1223-36. doi: 10.1163/1568562041753025.
In order to enhance the mechanical performances of hyaluronic acid (HA) without compromising its biological activity, HA has been interpenetrating with a fibrillar collagen scaffold. The semi-interpenetrating materials were obtained by mixing HA with different molecular weight and a pepsin-solubilized collagen (atelocollagen) solution, and then by inducing collagen fibrillogenesis. Results indicate that molecular weight of HA significantly influences the mechanical properties of the semi-interpenetrating materials and more specifically stronger material results from the use of low-molecular-weight (LMW) HA. According to the dynamic mechanical data the composite collagen-LMW HA has a higher elastic modulus than collagen, whereas the opposite is true for the high-molecular-weight (HMW) HA. This result highlights the role of specific interactions that occur between collagen and HA during the gel formation in controlling the network mechanical stability. LMW HA may, probably, interact more strongly with collagen during the fibrillogenesis process than HMW HA due to the higher mobility of the chains and the weaker homologous interactions. Moreover, morphological observations showed that LMW HA is intimately interdispersed within the collagen network and completely coated the fibrils, which act as mechanical support.
23Protein-mediated gelation and nano-scale assembly of unfunctionalized hyaluronic acid and chondroitin sulfate.PubMed
Anthony Tabet, June Y Park, Jarrod Shilts, et al.
F1000Res. 2018 Nov 20;7:1827. doi: 10.12688/f1000research.16929.3. eCollection 2018.
Hyaluronic acid (HA) is a major component of the extracellular matrix (ECM) in the central nervous system and the only purely supramolecular glycosaminoglycan. Much focus has been given to using this high molecular weight polysaccharide for tissue engineering applications. In most studies, the backbone of HA is functionalized with moieties that can facilitate network formation through physical self-assembly, or covalent crosslinking (e.g. photo-catalyzed) at concentrations where the polysaccharide does not gel on its own. However, these crosslinks often utilize functional groups not found in biological tissues. Oscillatory rheology, dynamic light scattering, and scanning electron microscopy were used to study albumin/HA structures. Dynamic light scattering and transmission electron microscopy were used to study albumin/chondroitin sulfate (CS) structures. UV-vis spectroscopy was used to demonstrate the potential for using protein-polymer blends as an ECM-mimetic model to study transport of small molecules. We examine the intermolecular interactions of two major glycosaminoglycans found in the human brain, HA and the lower molecular weight CS, with the model protein albumin. We report the properties of the resulting micro- and nano materials. Our albumin/HA systems formed gels, and albumin/CS systems formed micro- and nanoparticles. These systems are formed from unfunctionalized polysaccharides, which is an attractive and simple method of forming HA hydrogels and CS nanoparticles. We also summarize the concentrations of HA and CS found in various mammalian brains, which could potentially be useful for biomimetic scaffold development. Simple preparation of commercially available charged biomacromolecules results in interesting materials with structures at the micron and nanometer length-scales. Such materials may have utility in serving as cost-effective models of nervous system electrostatic interactions and as in vitro drug release and model system for ECM transport studies.
24Intra-articular Hyaluronic Acid Treatments for Knee Osteoarthritis: A Systematic Review of Product Properties.PubMed
Eric Ferkel, Ajay Manjoo, Damion Martins, et al.
Cartilage. 2023 Dec;14(4):424-432. doi: 10.1177/19476035231154530. Epub 2023 Jun 14.
INTRODUCTION: There are many intra-articular hyaluronic acid (IA-HA) products on the market that have known intrinsic differences in molecular size, source, and structure. The current review summarizes existing evidence describing and assessing these differences, while also identifying whether these differences have an impact on clinical outcomes. METHODS: This systematic review summarized all literature that specifically addresses IA-HA product differences. Included studies summarized basic science and mechanism of action comparisons of IA-HA product differences, or systematic reviews that assess differences in clinical outcomes between IA-HA product differences. RESULTS: A total of 20 investigations assessed basic science differences between IA-HA products, while 20 investigations provided assessments of the clinical outcome differences between IA-HA product characteristics. The published basic science literature provided a differentiation between low molecular weight (LMW) and high molecular weight (HMW) HA with regard to changes within the synovial fluid, driven by the interactions that these molecules have with receptors in the joint space. These differences in receptor interaction manifest within clinical outcomes, as meta-analyses comparing pain relief after IA-HA suggest that pain reduction is superior in patients who receive HMW HA as opposed to LMW HA. CONCLUSION: This review highlights differences between IA-HA characteristics, and how important the molecular weight, derivation of the product, and structure are to variances in reported clinical outcomes to treat osteoarthritis (OA) of the knee. HMW IA-HAs have shown greater efficacy compared to the alternative of LMW products, while avian-derived and cross-linked products have potentially demonstrated an increase in inflammatory events over non-avian-derived, non-cross-linked HAs.
25Low Molecular Weight Hyaluronic Acid (500-730 Kda) Injections in Tendinopathies-A Narrative Review.PubMed
Antonio Frizziero, Filippo Vittadini, Davide Bigliardi, et al.
J Funct Morphol Kinesiol. 2021 Dec 29;7(1):3. doi: 10.3390/jfmk7010003.
Tendinopathies are common causes of pain and disability in general population and athletes. Conservative treatment is largely preferred, and eccentric exercise or other modalities of therapeutic exercises are recommended. However, this approach requests several weeks of consecutive treatment and could be discouraging. In the last years, injections of different formulations were evaluated to accelerate functional recovery in combination with usual therapy. Hyaluronic acid (HA) preparations were proposed, in particular LMW-HA (500-730 kDa) for its unique molecular characteristics in favored extracellular matrix homeostasis and tenocyte viability. The purpose of our review is to evaluate the state-of-the-art about the role of 500-730 kDa in tendinopathies considering both preclinical and clinical findings and encourage further research on this emerging topic.
26Action of Hyaluronic Acid as a Damage-Associated Molecular Pattern Molecule and Its Function on the Treatment of Temporomandibular Disorders.PubMed
Natália Dos Reis Ferreira, Carolina Kaminski Sanz, Aline Raybolt, et al.
Front Pain Res (Lausanne). 2022 Mar 18;3:852249. doi: 10.3389/fpain.2022.852249. eCollection 2022.
The temporomandibular joint is responsible for fundamental functions. However, mechanical overload or microtraumas can cause temporomandibular disorders (TMD). In addition to external factors, it is known that these conditions are involved in complex biological mechanisms, such as activation of the immune system, activation of the inflammatory process, and degradation of extracellular matrix (ECM) components. The ECM is a non-cellular three-dimensional macromolecular network; its most studied components is hyaluronic acid (HA). HA is naturally found in many tissues, and most of it has a high molecular weight. HA has attributed an essential role in the viscoelastic properties of the synovial fluid and other tissues. Additionally, it has been shown that HA molecules can contribute to other mechanisms in the processes of injury and healing. It has been speculated that the degradation product of high molecular weight HA in healthy tissues during injury, a low molecular weight HA, may act as damage-associated molecular patterns (DAMPs). DAMPs are multifunctional and structurally diverse molecules that play critical intracellular roles in the absence of injury or infection. However, after cellular damage or stress, these molecules promote the activation of the immune response. Fragments from the degradation of HA can also act as immune response activators. Low molecular weight HA would have the ability to act as a pro-inflammatory marker, promoting the activation and maturation of dendritic cells, the release of pro-inflammatory cytokines such as interleukin 1 beta (IL-1β), and tumor necrosis factor α (TNF-α). It also increases the expression of chemokines and cell proliferation. Many of the pro-inflammatory effects of low molecular weight HA are attributed to its interactions with the activation of toll-like receptors (TLRs 2 and 4). In contrast, the high molecular weight HA found in healthy tissues would act as an anti-inflammatory, inhibiting cell growth and differentiation, decreasing the production of inflammatory cytokines, and reducing phagocytosis by macrophages. These anti-inflammatory effects are mainly attributed to the interaction of high-weight HA with the CD44 receptor. In this study, we review the action of the HA as a DAMP and its functions on pain control, more specifically in orofacial origin (e.g., TMD).
27[Hyaluronan-mediated regulation of inflammation].PubMed
Rafał Krasiński, Henryk Tchórzewski
Postepy Hig Med Dosw (Online). 2007 Nov 19;61:683-9.
Hyaluronan is high-molecular-weight biopolymer. Its linear structure is created by repeating disaccharide units. A single unit is composed of N-acetyl-D-glucosamine and D-glucuronic acid. Hyaluronan is the main component of the extracellular matrix. Apart from its structural role, hyaluronan can influence cell proliferation, differentiation, and migration, angiogenesis, as well as inflammation and immune cell function. During inflammation, high-molecular-weight hyaluronan is broken down under the influence of free radicals and enzymes to smaller fragments. Numerous literature data show that the effect of haluronan on immune cells is closely dependent on its molecular mass. High-molecular-weight hyaluronan can participate in restraining inflammation, while the low-molecular-weight form possesses a proinflammatory effect and activates immune cells. Through interaction with surface receptors (CD44, RHAMM, TLR4), hyaluronan fragments stimulate immune cells and enhance cytokine and reactive oxygen species production as well as other factors participating in inflammation. Hyaluronate can thus be an important regulator of the inflammatory process. Low-molecular-weight fragments deliver signal about tissue damage and mobilize immune cells, while the high-molecular-form suppresses immune cell function and prevents excessive exacerbation of inflammation.
28Anti-Inflammatory Effects of Intra-Articular Hyaluronic Acid: A Systematic Review.PubMed
Roy Altman, Asheesh Bedi, Ajay Manjoo, et al.
Cartilage. 2019 Jan;10(1):43-52. doi: 10.1177/1947603517749919. Epub 2018 Feb 11.
OBJECTIVE: Osteoarthritis (OA) is one of the leading causes of disability in the adult population. Common nonoperative treatment options include nonsteroidal anti-inflammatory drugs (NSAIDs), intra-articular corticosteroids, and intra-articular injections of hyaluronic acid (HA). HA is found intrinsically within the knee joint providing viscoelastic properties to the synovial fluid. HA therapy provides anti-inflammatory relief through a number of different pathways, including the suppression of pro-inflammatory cytokines and chemokines. METHODS: We conducted a systematic review to summarize the published literature on the anti-inflammatory properties of hyaluronic acid in osteoarthritis. Included articles were categorized based on the primary anti-inflammatory responses described within them, by the immediate cell surface receptor protein assessed within the article, or based on the primary theme of the article. Key findings aimed to describe the macromolecules and inflammatory-mediated responses associated with the cell transmembrane receptors. RESULTS: Forty-eight articles were included in this systematic review that focused on the general anti-inflammatory effects of HA in knee OA, mediated through receptor-binding relationships with cluster determinant 44 (CD44), toll-like receptor 2 (TLR-2) and 4 (TLR-4), intercellular adhesion molecule-1 (ICAM-1), and layilin (LAYN) cell surface receptors. Higher molecular weight HA (HMWHA) promotes anti-inflammatory responses, whereas short HA oligosaccharides produce inflammatory reactions. CONCLUSIONS: Intra-articular HA is a viable therapeutic option in treating knee OA and suppressing inflammatory responses. HMWHA is effective in suppressing the key macromolecules that elicit the inflammatory response by short HA oligosaccharides.
29High and low molecular weight hyaluronic acid differentially influence macrophage activation.PubMed
Jamie E Rayahin, Jason S Buhrman, Yu Zhang, et al.
ACS Biomater Sci Eng. 2015 Jul 13;1(7):481-493. doi: 10.1021/acsbiomaterials.5b00181.
Macrophages exhibit phenotypic diversity permitting wide-ranging roles in maintaining physiologic homeostasis. Hyaluronic acid, a major glycosaminoglycan of the extracellular matrix, has been shown to have differential signaling based on its molecular weight. With this in mind, the main objective of this study was to elucidate the role of hyaluronic acid molecular weight on macrophage activation and reprogramming. Changes in macrophage activation were assessed by activation state selective marker measurement, specifically quantitative real time polymerase chain reaction, and cytokine enzyme-linked immunoassays, after macrophage treatment with differing molecular weights of hyaluronic acid under four conditions: the resting state, concurrent with classical activation, and following inflammation involving either classically or alternatively activated macrophages. Regardless of initial polarization state, low molecular weight hyaluronic acid induced a classically activated-like state, confirmed by up-regulation of pro-inflammatory genes, including , , , and , and enhanced secretion of nitric oxide and TNF-α. High molecular weight hyaluronic acid promoted an alternatively activated-like state, confirmed by up regulation of pro-resolving gene transcription, including , , and , and enhanced arginase activity. Overall, our observations suggest that macrophages undergo phenotypic changes dependent on molecular weight of hyaluronan that correspond to either (1) pro-inflammatory response for low molecular weight HA or (2) pro-resolving response for high molecular weight HA. These observations bring significant further understanding of the influence of extracellular matrix polymers, hyaluronic acid in particular, on regulating the inflammatory response of macrophages. This knowledge can be used to guide the design of HA-containing biomaterials to better utilize the natural response to HAs.
30Hyaluronic Acid in Inflammation and Tissue Regeneration.PubMed
Malgorzata Litwiniuk, Alicja Krejner, Marcus S Speyrer, et al.
Wounds. 2016 Mar;28(3):78-88.
Hyaluronic acid (HA), the main component of extracellular matrix, is considered one of the key players in the tissue regeneration process. It has been proven to modulate via specific HA receptors, inflammation, cellular migration, and angiogenesis, which are the main phases of wound healing. Studies have revealed that most HA properties depend on its molecular size. High molecular weight HA displays anti-inflammatory and immunosuppressive properties, whereas low molecular weight HA is a potent proinflammatory molecule. In this review, the authors summarize the role of HA polymers of different molecular weight in tissue regeneration and provide a short overview of main cellular receptors involved in HA signaling. In addition, the role of HA in 2 major steps of wound healing is examined: inflammation and the angiogenesis process. Finally, the antioxidative properties of HA are discussed and its possible clinical implication presented.
31Effects of 4-methylumbelliferone and high molecular weight hyaluronic acid on the inflammation of corneal stromal cells induced by LPS.PubMed
Fang Li, Peng Hao, Guangjie Liu, et al.
Graefes Arch Clin Exp Ophthalmol. 2017 Mar;255(3):559-566. doi: 10.1007/s00417-016-3561-1. Epub 2016 Dec 6.
PURPOSE: To investigate the effects of hyaluronic acid (HA) on the inflammation of corneal fibroblasts induced by lipopolysaccharide (LPS). METHODS: Primary rabbit corneal keratocytes were isolated with collagenase. The keratocytes were cultured in a serum-containing medium to induce corneal fibroblasts, which represented the wound repair phenotype of corneal keratocytes. Corneal fibroblasts were treated with LPS with or without 4-methylumbelliferone (4-MU) / high molecular weight hyaluronic acid (HMWHA). The gene expression was evaluated via real-time PCR, immunofluorescence, and western blot. The release of inflammatory cytokines and HA was determined by ELISA. RESULTS: Three types of hyaluronan synthase (HAS) were detected in corneal fibroblasts. LPS stimulation caused the up-regulation of HAS1 and HAS2 expression in corneal fibroblasts. LPS-induced HAS2 expression was significantly inhibited by 4-MU, and accompanied by decreased HA release by the corneal fibroblasts. In the corneal fibroblasts, 4-MU reduced the LPS-stimulated up-regulation of inflammatory cytokines including IL-1, IL-6, IL-8, TNF-α, and also attenuated the LPS-induced up-regulation of inflammatory related receptors including TLR2, TLR4, CD44, and CXCR1. HMWHA treatment resulted in a significant decline in the expression of IL-6, IL-8, TLR4, and CXCR1 responded to LPS stimulation. Consistent with mRNA expression of level, the up-regulation of the release of IL-6 and IL-8 induced by LPS in corneal fibroblasts was significantly attenuated by 4-MU and HMWHA. The LPS-induced expression of IL-8 and its receptor CXCR1 at both the mRNA and protein level were significantly attenuated by 4-MU and HMWHA. CONCLUSION: The inhibitor of HA synthesis 4-MU, and HMWHA successfully reduced LPS-induced inflammation in corneal fibroblasts. The mechanism might be via the inhibition of LPS-induced TLR4 up-regulation.
32The stimulation of adenosine 2A receptor reduces inflammatory response in mouse articular chondrocytes treated with hyaluronan oligosaccharides.PubMed
Giuseppe M Campo, Angela Avenoso, Angela D'Ascola, et al.
Matrix Biol. 2012 Jul;31(6):338-51. doi: 10.1016/j.matbio.2012.07.001. Epub 2012 Jul 13.
The adenosine 2A receptor (A(2A)R) is greatly involved in inflammation pathologies such as rheumatoid arthritis. By interacting with A(2A)R, the purine nucleoside adenosine acts as a potent endogenous inhibitor of the inflammatory process in a variety of tissues. Hyaluronan (HA) fragments act to prime inflammation via CD44 and the toll-like receptor 4 (TLR-4). The aim of this study was to investigate whether the inhibition/stimulation of A(2A)R modulates the inflammation cascade primed by small HA fragments in mouse articular chondrocytes. 6-mer HA treatment induced up-regulation of CD44, TLR4 and A(2A)R mRNA expression and the related protein levels, and NF-kB activation, that in turn increased TNF-α, IL-1β, and IL-6 and production. Treatment with a selective (2)A adenosine receptor agonist (2-phenylaminoadenosine) enhanced A(2A)R increase, as well as the inhibition of CD44 and TLR4 activity using two specific antibodies abolished up-regulation of CD44 and TLR4, and significantly reduced, especially by antibody inhibition, NF-kB activation and pro-inflammatory cytokine production. Furthermore, the exposure of chondrocytes to A(2A)R specific interference mRNA (A(2A)R siRNA) enhanced HA 6-mer induced NF-kB activation and inflammatory cytokine increase. Finally, the use of an exchange protein activated by cAMP (EPAC) siRNA and a specific PKA inhibitor showed a predominant EPAC involvement in the mediation of the anti-inflammatory activity exerted by A(2A)R stimulation. These data suggest that HA depolymerization occurring during inflammation contributes to priming of the inflammatory cascade, while endogenous adenosine, by exerting anti-inflammatory response via A(2A)R, could be a modulatory mechanism that attempts to attenuate the inflammation process.
33Low-Molecular-Weight Hyaluronic Acid Contributes to Noise-Induced Cochlear Inflammation.PubMed
Guoping Zhang, Yiling Gao, Zhen Zhao, et al.
Audiol Neurootol. 2023;28(5):380-393. doi: 10.1159/000530280. Epub 2023 May 9.
INTRODUCTION: Our previous work indicated that the activation of the Toll-like receptor (TLR) 4 signaling pathway contributed to noise-induced cochlear inflammation. Previous studies have reported that low-molecular-weight hyaluronic acid (LMW-HA) accumulates during aseptic trauma and promotes inflammation by activating the TLR4 signaling pathway. We hypothesized that LMW-HA or enzymes synthesizing or degrading HA might be involved in noise-induced cochlear inflammation. METHODS: The present study included two arms. The first arm was the noise exposure study, in which TLR4, proinflammatory cytokines, HA, hyaluronic acid synthases (HASs), and hyaluronidases (HYALs) in the cochlea as well as auditory brainstem response (ABR) thresholds were measured before and after noise exposure. The second arm was analysis of HA delivery-induced reactions, in which control solution, high-molecular-weight HA (HMW-HA), or LMW-HA was delivered into the cochlea by cochleostomy or intratympanic injection. Then, the ABR threshold and cochlear inflammation were measured. RESULTS: After noise exposure, the expression of TLR4, proinflammatory cytokines, HAS1, and HAS3 in the cochlea significantly increased over the 3rd to 7th day post-noise exposure (PE3, PE7). The expression of HYAL2 and HYAL3 dramatically decreased immediately after noise exposure, gradually increased thereafter to levels significantly greater than the preexposure level on PE3, and then rapidly returned to the preexposure level on PE7. The expression of HA, HAS2, and HYAL1 in the cochlea remained unchanged after exposure. After cochleostomy or intratympanic injection, both the hearing threshold shifts and the expression of TLR4, TNF-α, and IL-1β in the cochleae of the LMW-HA group were obviously greater than those of the control group and HMW-HA group. The expression of proinflammatory cytokines in the LMW-HA and control groups on the 7th day (D7) after cochleostomy tended to increase compared to that on the 3rd day (D3), whereas levels in the HMW-HA group tended to decrease on D7 compared to D3. CONCLUSION: HAS1, HAS3, HYAL2, and HYAL3 in the cochlea are involved in acoustic trauma-induced cochlear inflammation through the potential proinflammatory function of LMW-HA.
34The anti-inflammatory activity of specific-sized hyaluronic acid oligosaccharides.PubMed
Wenwei Han, Youjing Lv, Yutong Sun, et al.
Carbohydr Polym. 2022 Jan 15;276:118699. doi: 10.1016/j.carbpol.2021.118699. Epub 2021 Sep 24.
Severe acute inflammatory conditions may cause tissue damage, sepsis, and death. As a critical component of the extracellular matrix, hyaluronic acid (HA) has been reported to possess pro- and anti-inflammatory properties via Toll-like receptors (TLRs). In this study, we prepared different sizes and structures of HA oligosaccharides and derivatives and investigated the effects on inflammation in vitro and in vivo. Our results showed that HA tetra-saccharide was the minimum fragment to enhance inflammation, whereas HA disaccharide competitively blocked TLR4-dependent inflammation. The enzymatic HA disaccharide (ΔHA2) inhibited lipopolysaccharide (LPS)-induced inflammation. Based on structure-activity relationship analysis, we observed that anti-inflammatory activity depended on HAs polymerization degree, acetyl group, and configuration. In addition, we demonstrated that ΔHA2 reduced LPS-induced pro-inflammatory cytokines production in vivo. ΔHA2, a native metabolite of HA polysaccharides, may have a potential role against LPS-mediated inflammatory diseases.
35Hyaluronan inhibits prostaglandin E2 production via CD44 in U937 human macrophages.PubMed
Tadashi Yasuda
Tohoku J Exp Med. 2010 Mar;220(3):229-35. doi: 10.1620/tjem.220.229.
Prostaglandin E(2) (PGE(2)) is one of the key mediators of inflammation in affected joints of rheumatoid arthritis (RA). Intra-articular injection of high molecular weight hyaluronan (HA) into RA knee joints relieves arthritic pain. Although HA has been shown to inhibit PGE(2) production in cytokine-stimulated synovial fibroblasts, it remains unclear how HA suppresses PGE(2) production in activated cells. Furthermore, HA effect on macrophages has rarely been investigated in spite of their contribution to RA joint pathology. This study was aimed to investigate the inhibitory mechanism of HA on lipopolysaccharide (LPS)-stimulated PGE(2) production in U937 human macrophages. Stimulation of U937 macrophages with LPS enhanced PGE(2) production in association with increased protein levels of cyclooxygenase-2 (COX-2). Pretreatment with HA of 2,700 kDa resulted in suppression of the LPS-mediated induction of COX-2, leading to a decrease in PGE(2) production. Likewise, the LPS-stimulated PGE(2) production was inhibited by the pretreatment with a specific COX2 inhibitor, NS-398, or a specific inhibitor of nuclear factor (NF)-kappaB, BAY11-7085. HA also decreased the degree of phosphorylation and nuclear translocation of NF-kappaB enhanced by LPS. Fluorescence cytochemistry demonstrated that HA bound to CD44, the principal HA receptor, on U937 macrophages. Anti-CD44 antibody reversed the inhibitory effects of HA on the LPS-mediated increase in PGE(2) production, COX-2 induction, and activation of NF-kappaB. These results indicate that HA suppresses the LPS-stimulated PGE(2) production via CD44 through down-regulation of NF-kappaB. Administration of HA into RA joints may decrease PGE(2) production by activated macrophages, which could result in improvement of arthritic pain.
36High molecular weight hyaluronic acid inhibits advanced glycation endproduct-induced NF-kappaB activation and cytokine expression.PubMed
A Neumann, R Schinzel, D Palm, et al.
FEBS Lett. 1999 Jun 25;453(3):283-7. doi: 10.1016/s0014-5793(99)00731-0.
Advanced glycation endproducts (AGEs), which accumulate on long-lived proteins and protein deposits (amyloids), induce the expression of proinflammatory cytokines through NF-kappaB-dependent pathways. Hyaluronic acid with a molecular weight above 1.2 MDa (HMW-HA) inhibits the AGE-induced activation of the transcription factor NF-kappaB and the NF-kappaB-regulated cytokines interleukin-1alpha, interleukin-6 and tumor necrosis factor-alpha. Since the molecular weight of hyaluronic acid in humans decreases with age and under conditions of oxidative stress, it is likely that the protective effect of HMW-HA against AGE-induced cellular activation is lost at sites of chronic inflammation and in older age.
37Anti-inflammatory effects of differential molecular weight Hyaluronic acids on UVB-induced calprotectin-mediated keratinocyte inflammation.PubMed
Liuying Hu, Satoshi Nomura, Yasunari Sato, et al.
J Dermatol Sci. 2022 Jul;107(1):24-31. doi: 10.1016/j.jdermsci.2022.06.001. Epub 2022 Jun 9.
BACKGROUND: The biological functions of Hyaluronic acid are related to its molecular weight and binding to its receptor, Toll-like receptor4 (TLR4) or CD44. Recent studies have shown that low-molecular-weight Hyaluronic acid (LMW-HA) exhibits proinflammatory effects, while high-molecular-weight Hyaluronic acid (HMW-HA) functions as an anti-inflammatory factor. UVB-induced epidermal inflammation is mainly mediated by endogenous molecules, such as damage-associated molecular patterns (DAMPs), that cause severe skin damage by activating TLR signaling pathways. OBJECTIVE: Since both LMW- and HMW-HA have inhibitory functions on TLR-mediated macrophage inflammation, HA is assumed to suppress UVB-induced DAMP-mediated inflammation in the skin. In this study, both Ultra- low-molecular-weight Hyaluronic acid (uLMW-HA) and HMW-HA were found to inhibit UVB-induced keratinocyte inflammation. METHODS: HaCaT cells were treated with medium containing Hyaluronic acid at the appropriate concentration after 15 mJ/cm irradiation. Secreted protein levels were determined with ELISA kits. Expression levels of proteins downstream of TLR4 were detected by Simple Western system. RESULTS: By competitively binding to TLR4, uLMW-HA downregulated Calprotectin-induced TRAF6 expression, which might be the direct process by which uLMW-HA decreased UVB-induced IL-6 secretion. Reduced CD44 variant (CD44v) expression in keratinocytes attenuated the inhibitory effect of both uLMW-HA and HMW-HA on UVB-induced inflammation, which indicated the involvement of CD44v in HA-regulated anti-inflammatory activity. CONCLUSION: Overall, this research indicates that Hyaluronic acid is more than a moisturizer; it is also a biologically effective material that can prevent the excessive skin inflammation caused in daily life, especially in the late stages after sunburn.
38A 3D "" Model to Study Hyaluronan Effect in Nasal Epithelial Cell Line Exposed to Double-Stranded RNA Poly(I:C).PubMed
Giusy Daniela Albano, Anna Bonanno, Daniela Giacomazza, et al.
Biomol Ther (Seoul). 2020 May 1;28(3):272-281. doi: 10.4062/biomolther.2019.126.
Environmental agents, including viral and bacterial infectious agents, are involved in the alteration of physicochemical and biological parameters in the nasal epithelium. Hyaluronan (HA) has an important role in the regulation of tissue healing properties. High molecular weight HA (HMW-HA) shows greater anti-inflammatory responses than medium molecular weight HA (MMW-HA) and low molecular weight HA (LMW-HA). We investigated the effect of HMW-HA, MMW-HA and LMW-HA on the regulation of physicochemical and biological parameters in an "" model that might mimic viral infections of the nasal epithelium. Human nasal epithelial cell line RPMI2650 was stimulated with double-stranded RNA (dsRNA) Poly(I:C) for 5 days in air-liquid-interface (ALI) culture (3D model of airway tissue). dsRNA Poly(I:C) treatment significantly decreased transepithelial electrical resistance (TEER) in the stratified nasal epithelium of RPMI2650 and increased pH values, rheological parameters (elastic G' and viscous G"), and Muc5AC and Muc5B production in the apical wash of ALI culture of RPMI2650 in comparison to untreated cells. RPMI2650 treated with dsRNA Poly(I:C) in the presence of HMW-HA showed lower pH values, Muc5AC and Muc5B production, and rheological parameters, as well as increased TEER values in ALI culture, compared to cells treated with Poly(I:C) alone or pretreated with LMW-HA and MMW-HA. Our 3D "" model of epithelium suggests that HMW-HA might be a coadjuvant in the pharmacological treatment of viral infections, allowing for the control of some physicochemical and biological properties affecting the epithelial barrier of the nose during infection.
39Hyaluronic acid synthase 2 dysfunction exacerbates elastase-induced neutrophilic airway inflammation and emphysema in mice.PubMed
Kengo Nishino, Takumi Kiwamoto, Masashi Matsuyama, et al.
Front Immunol. 2025 Oct 14;16:1683385. doi: 10.3389/fimmu.2025.1683385. eCollection 2025.
Chronic obstructive pulmonary disease (COPD) is a progressive inflammatory lung disorder primarily caused by prolonged exposure to harmful substances, such as cigarette smoke. Hyaluronic acid synthase 2 (HAS2) synthesizes high-molecular-weight hyaluronic acid (HMW-HA), which has anti-inflammatory properties. Previous studies have revealed that HAS2 dysfunction may exacerbate COPD. However, the specific impact of HAS2 on pulmonary emphysema progression remains unclear. Therefore, this study examined whether HAS2 dysfunction worsens airway inflammation and emphysema in a mouse COPD model. heterozygous-deficient ( ) mice and their wild-type (WT) littermates were evaluated using a porcine pancreatic elastase (PPE)-induced COPD model. After the administration of PPE, the mice exhibited a significant increase in total cell and neutrophil counts in the bronchoalveolar lavage fluid samples compared with the WT mice. Further, the mice presented with enhanced emphysema development on histological analyses, with higher mean linear intercept values relative to the WT mice. The PPE-stimulated mice also had increased G-CSF levels and tumor growth factor-beta (TGF-β) attenuation in the lungs. RNA-sequencing analysis revealed that PPE stimulation promoted the synthesis of HMW-HA and TGF-β signaling. Gene Ontology analysis using mice-specific differentially expressed genes showed that the genes associated with the pathways that promote the negative regulation of the TGF-β receptor signaling pathway were activated after the administration of PPE. Therefore, dysfunction exacerbates neutrophilic airway inflammation and emphysema, thereby underscoring the protective role of HAS2 in a PPE-induced emphysema model. The exacerbated response may involve G-CSF- and TGF-β-related signaling pathways. These findings may contribute to the development of novel therapeutic strategies for COPD management.
40Molecular Mechanisms and Therapeutic Role of Intra-Articular Hyaluronic Acid in Osteoarthritis: A Precision Medicine Perspective.PubMed
Wojciech Glinkowski, Dariusz Śladowski, Wiesław Tomaszewski, et al.
J Clin Med. 2025 Apr 8;14(8):2547. doi: 10.3390/jcm14082547.
: Osteoarthritis (OA) is a degenerative joint disease characterized by progressive cartilage breakdown, synovial inflammation, and pain, which leads to significant disability. IAHA is widely used because of its viscoelastic properties, which restore synovial fluid homeostasis and reduce symptoms. However, emerging evidence suggests that IAHA exerts additional biological effects including chondroprotection, inflammatory modulation, oxidative stress reduction, and pain modulation, which may influence disease progression. : This narrative review examines the biological mechanisms underlying IAHA's role in OA management. The review explored IAHA's effects on synovial fluid viscoelasticity, inflammatory cytokine modulation, cartilage preservation, oxidative stress regulation, and pain pathways, emphasizing the influence of molecular weight variations on therapeutic efficacy. Additionally, this review evaluates IAHA's integration into multimodal treatment strategies, its potential disease-modifying effects, and future directions for personalized treatment approaches. : A comprehensive literature review was conducted using PubMed, Cochrane Library, EMBASE, Scopus, and Web of Science for studies published between January 2000 and March 2024. The search focused on IAHA's molecular, cellular, and biochemical effects in OA and clinical findings assessing its impact on joint function, pain relief, and disease progression. : IAHA improves synovial fluid lubrication, reduces proinflammatory cytokines (IL-1β, TNF-α), inhibits matrix metalloproteinases (MMPs), scavenges reactive oxygen species (ROS), and modulates nociceptive pathways. High-molecular-weight IAHA demonstrates superior efficacy in advanced OA, while low-molecular-weight formulations may be better suited for early-stage disease. Although IAHA's symptom relief is comparable to corticosteroids and NSAIDs, its favorable safety profile and emerging disease-modifying potential support its long-term use in OA management. : IAHA represents a multifaceted therapeutic approach bridging symptomatic relief and regenerative strategies. While long-term efficacy, optimal administration protocols, and patient-specific responses remain subjects of ongoing research, refining treatment selection criteria, dosing regimens, and combination strategies may enhance clinical outcomes. Future studies should explore biomarker-driven approaches, standardize treatment protocols, and assess IAHA's synergy with regenerative medicine to optimize its role in OA management.
41Deciphering Drug Resistance: Investigating the Emerging Role of Hyaluronan Metabolism and Signaling and Tumor Extracellular Matrix in Cancer Chemotherapy.PubMed
Daiana L Vitale, Arianna Parnigoni, Manuela Viola, et al.
Int J Mol Sci. 2024 Jul 11;25(14):7607. doi: 10.3390/ijms25147607.
Hyaluronan (HA) has gained significant attention in cancer research for its role in modulating chemoresistance. This review aims to elucidate the mechanisms by which HA contributes to chemoresistance, focusing on its interactions within the tumor microenvironment. HA is abundantly present in the extracellular matrix (ECM) and binds to cell-surface receptors such as CD44 and RHAMM. These interactions activate various signaling pathways, including PI3K/Akt, MAPK, and NF-κB, which are implicated in cell survival, proliferation, and drug resistance. HA also influences the physical properties of the tumor stroma, enhancing its density and reducing drug penetration. Additionally, HA-mediated signaling contributes to the epithelial-mesenchymal transition (EMT), a process associated with increased metastatic potential and resistance to apoptosis. Emerging therapeutic strategies aim to counteract HA-induced chemoresistance by targeting HA synthesis, degradation, metabolism, or its binding to CD44. This review underscores the complexity of HA's role in chemoresistance and highlights the potential for HA-targeted therapies to improve the efficacy of conventional chemotherapeutics.
42Hyaluronan up-regulates growth and invasion of trophoblasts in an autocrine manner via PI3K/AKT and MAPK/ERK1/2 pathways in early human pregnancy.PubMed
R Zhu, Y-H Huang, Y Tao, et al.
Placenta. 2013 Sep;34(9):784-91. doi: 10.1016/j.placenta.2013.05.009. Epub 2013 Jun 24.
INTRODUCTION: As one of the key molecules in the extracellular matrix in human conceptus, hyaluronan (HA) has been receiving particular attention. Here, we have investigated the expression and regulation of different molecular weight HA on the biological behaviors of primary human trophoblasts during the first trimester of pregnancy. METHODS: The expression of HA and HA synthetase (HAS) by human first trimester trophoblasts was analyzed in placentae from normal pregnancy or miscarriage by immunochemistry and real-time RT-PCR, respectively. ELISA was used to measure the secretion of HA by primary trophoblasts. The effects of HA on the proliferation, apoptosis and invasiveness of trophoblasts were examined. We also investigated the signaling pathways involved in HA activation in human trophoblasts. RESULTS: The higher HAS2 expression and HA secretion were observed in normal villi than that of miscarriage, and the primary trophoblasts secreted HA continuously. High molecular weight HA (HMW-HA) and medium molecular weight HA (MMW-HA) promoted proliferation and invasiveness while inhibited apoptosis of trophoblasts. However, low molecular weight HA (LMW-HA) had no obvious effect on the growth or invasiveness of human trophoblasts. In addition, HMW-HA showed more efficiently than MMW-HA on the growth while MMW-HA displayed a more obvious effect on the invasiveness of trophoblasts than HMW-HA. HMW-HA activated PI3K/AKT and MAPK/ERK1/2 signaling pathways in trophoblasts. Blocking PI3K/AKT or MAPK/ERK1/2 signaling inhibited the HA-upregulated growth and invasiveness of human trophoblasts. CONCLUSION: Our results suggest that higher level and greater molecular mass of HA can promote trophoblast growth and invasion in an autocrine manner, which was beneficial to placentation and maintenance of human early pregnancy.
43Hyaluronic acid: More than a carrier, having an overpowering extracellular and intracellular impact on cancer.PubMed
Medha Bhattacharyya, Heena Jariyal, Akshay Srivastava
Carbohydr Polym. 2023 Oct 1;317:121081. doi: 10.1016/j.carbpol.2023.121081. Epub 2023 Jun 3.
Hyaluronic acid (HA), also named hyaluronan, is an omnipresent component of the tissue microenvironment. It is extensively used to formulate targeted drug delivery systems for cancer. Although HA itself has pivotal influences in various cancers, its calibers are somewhat neglected when using it as delivering platform to treat cancer. In the last decade, multiple studies revealed roles of HA in cancer cell proliferation, invasion, apoptosis, and dormancy through pathways like mitogen-activated protein kinase-extracellular signal-regulated kinase (MAPK/ERK), P, and nuclear factor kappa-light chain-enhancer of activated B cells (NFκB). A more fascinating fact is that the distinct molecular weight (MW) of HA exerts disparate effects on the same type of cancer. Its overwhelming use in cancer therapy and other therapeutic products make collective research on the sundry impact of it on various types of cancer, an essential aspect to be considered in all of these domains. Even the development of new therapies against cancer needed meticulous studies on HA because of its divergence of activity based on MW. This review will provide painstaking insight into the extracellular and intracellular bioactivity of HA, its modified forms, and its MW in cancers, which may improve the management of cancer.
44Hyaluronic acid-dependent protection against alkali-burned human corneal cells.PubMed
Chieh-Lin Wu, Hsiu-Chuan Chou, Ji-Min Li, et al.
Electrophoresis. 2013 Feb;34(3):388-96. doi: 10.1002/elps.201200342. Epub 2013 Jan 6.
Hyaluronic acid (HA) is a high-molecular-weight glycosaminoglycan and extracellular matrix component that promotes cell proliferation. This study aimed to evaluate the effects of HA on alkali-injured human corneal epithelial cells in vitro, and to elucidate the mechanisms by which HA mediates corneal cell protection. A human corneal epithelial cell line (HCE-2) was treated with sodium hydroxide before incubation with low-molecular-weight HA (LMW-HA, 127 kDa) or high-molecular-weight HA (HMW-HA, 1525 kDa). A global proteomic analysis was then performed. Our data indicated that HA treatment protects corneal epithelial cells from alkali injury, and that the molecular weight of HA is a crucial factor in determining its effects. Only HMW-HA reduced NaOH-induced cytotoxic effects in corneal cells significantly and increased their migratory and wound healing ability. Results from 2D-DIGE and MALDI-TOF/TOF MS analyses indicated that HMW-HA modulates biosynthetic pathways, cell migration, cell outgrowth, and protein degradation to stimulate wound healing and prevent cell death. To our knowledge, our study is the first to report the possible mechanisms by which HMW-HA promotes repair in alkali-injured human corneal epithelial cells.
45Hyaluronic acid-dependent protection against UVB-damaged human corneal cells.PubMed
Ji-Min Li, Hsiu-Chuan Chou, Ssu-Han Wang, et al.
Environ Mol Mutagen. 2013 Jul;54(6):429-49. doi: 10.1002/em.21794. Epub 2013 Jun 27.
Within ultraviolet radiation, ultraviolet B (UVB) is the most energetic and damaging to humans. At the protein level, UVB irradiation downregulates the expression of antioxidant enzymes leading to the accumulation of reactive oxygen species (ROS). Due to lacking of a global analysis of UVB-modulated corneal proteome, we investigate in vitro the mechanism of UVB-induced corneal damage to determine whether hyaluronic acid (HA) is able to reduce UVB irradiation-induced injury in human corneal epithelial cells. Accordingly, human corneal epithelial cell lines (HCE-2) were irradiated with UVB, followed by incubation with low molecular weight HA (LMW-HA, 100 kDa) or high molecular weight HA (HMW-HA, 1,000 kDa) to investigate the physiologic protection of HMW-HA in UVB-induced corneal injury, and to perform a global proteomic analysis. The data demonstrated that HA treatment protects corneal epithelial cells in the UVB-induced wound model, and that the molecular weight of HA is a crucial factor. Only HMW-HA significantly reduces the UVB-induced cytotoxic effects in corneal cells and increases cell migration and wound-healing ability. In addition, proteomic analysis showed that HMW-HA might modulate cytoskeleton regulation, signal transduction, biosynthesis, redox regulation, and protein folding to stimulate wound healing and to prevent these UVB-damaged cells from cell death. Further studies evidenced membrane-associated progesterone receptor component 1 (mPR) and malate dehydrogenase (MDH2) play essential roles in protecting corneal cells from UVB irradiation. This study reports on UVB-modulated cellular proteins that might play an important role in UVB-induced corneal cell injury and show HMW-HA to be a potential substance for protecting corneal cells from UVB-induced injury.
46Effect of hyaluronic acid on the thermogelation and biocompatibility of its blends with methyl cellulose.PubMed
Laura Mayol, Daniela De Stefano, Francesca De Falco, et al.
Carbohydr Polym. 2014 Nov 4;112:480-5. doi: 10.1016/j.carbpol.2014.06.020. Epub 2014 Jun 17.
Aim of this work was to investigate the influence of hyaluronic acid (HA) molecular weight on the thermogelation and biocompatibility of its blends with methyl cellulose in view of a possible application in drug delivery and/or wound healing. We found out that it was possible to obtain MC/HA blends showing a rheological behavior typical of a viscous solution at 20 °C and of a weak gel at 37 °C only when blending MC with low molecular weight HA. Moreover, the blends containing low molecular weight HA did not affect human foreskin fetal fibroblasts viability, proliferation and migration. On the contrary, the cell incubation with high molecular weight HA resulted in a marked and significant reduction of cell viability, compared to control cells. Finally, the optimized blends, in terms of rheological properties and biocompatibility, proved to be able to control and prolong bovine serum albumin release by a combined mechanism of platform dissolution and drug diffusion.
47Impact of Hyaluronic Acid on the Cutaneous T-Cell Lymphoma Microenvironment: A Novel Anti-Tumor Mechanism of Bexarotene.PubMed
Tetsuya Ikawa, Emi Yamazaki, Ryo Amagai, et al.
Cancers (Basel). 2025 Jan 20;17(2):324. doi: 10.3390/cancers17020324.
BACKGROUND: Cutaneous T-cell lymphoma (CTCL) is a type of non-Hodgkin's lymphoma that primarily affects the skin, rich in hyaluronic acid (HA). HA is a component of the extracellular matrix in the dermis and likely affects the development of CTCL, but the mechanism is poorly understood. Here we show that low-molecular-weight HA (LMWHA) possibly exacerbates CTCL, and bexarotene, already used in CTCL treatment, decreases HA production. METHODS: We conducted immunohistochemistry, qRT-PCR, immunoblotting, and HA quantification using both mouse and human specimens to evaluate the impact of HA on CTCL. Additionally, we assessed the effect of bexarotene, which is already used for CTCL treatment, on HA metabolism. RESULTS: HA expression was higher in patients' serum and skin sections than in healthy controls. HA extracted from the skin of mice inoculated with tumors showed an increase in LMWHA. LMWHA increased lymphoma cell proliferation in vitro and accelerated tumor formation in mice in vivo. LMWHA also created a favorable environment for tumor cells by affecting fibroblasts, vascular endothelial cells, and tumor-associated macrophages. Thus, increased levels of HA, mainly LMWHA, exacerbate CTCL progression by affecting tumor cells and their microenvironment. Bexarotene treatment reduced the amount of total HA in murine tumor-inoculated skin, as well as the supernatant of cultured normal human dermal fibroblasts (NHDFs) and HuT78 cells. Detailed in vitro analyses showed that bexarotene treatment decreased HA synthase (HAS)1 and HAS2 expression in NHDFs and HAS1 and HAS3, and CEMIP expression in HuT78 cells. Chromatin immunoprecipitation assays revealed that bexarotene reduced retinoid X receptor-α binding to the and promoters in NHDFs. CONCLUSIONS: Bexarotene potentially exerts its anti-tumor effect by reducing HA levels through decreased expression of HAS. These findings provide new insights into the process of CTCL development and additional insights regarding bexarotene treatment.
48Hyaluronic acid in digestive cancers.PubMed
Ruo-Lin Wu, Lei Huang, Hong-Chuan Zhao, et al.
J Cancer Res Clin Oncol. 2017 Jan;143(1):1-16. doi: 10.1007/s00432-016-2213-5. Epub 2016 Aug 17.
PURPOSE: Hyaluronan (HA), an extracellular and peri-cellular glycosaminoglycan with a large molecular weight, plays an important role in cancer growth and metastasis. The aim of this study was to summarize the biological roles and regulation of HA and small HA fragments, and their metabolismn enzymes and receptors in human digestive cancers. METHODS: A systematic literature search mainly focusing on the biological roles of HA in the development and progression of human digestive cancers was performed using electronic databases. RESULTS: The correlation between HA accumulation and tumor progression has been shown in various digestive cancers. HA and HA fragment-tumor cell interaction could activate the downstream signaling pathways, promoting cell proliferation, adhesion, migration and invasion, and inducing angiogenesis, lymphangiogenesis, epithelial-mesenchymal transition, stem cell-like property, and chemoradioresistance in digestive cancers. CONCLUSIONS: A better insight into the mechanism of HA and HA fragment involvement in digestive cancer progression might be useful for the development of novel biomarkers and therapeutic strategies.
49Hyaluronan stimulates pancreatic cancer cell motility.PubMed
Xiao-Bo Cheng, Shiro Kohi, Atsuhiro Koga, et al.
Oncotarget. 2016 Jan 26;7(4):4829-40. doi: 10.18632/oncotarget.6617.
Hyaluronan (HA) accumulates in pancreatic ductal adenocarcinoma (PDAC), but functional significance of HA in the aggressive phenotype remains unknown. We used different models to investigate the effect of HA on PDAC cell motility by wound healing and transwell migration assay. Changes in cell motility were examined in 8 PDAC cell lines in response to inhibition of HA production by treatment with 4-methylumbelliferone (4-MU) and to promotion by treatment with 12-O-tetradecanoyl-phorbol-13-acetate (TPA) or by co-culture with tumor-derived stromal fibroblasts. We also investigated changes in cell motility by adding exogenous HA. Additionally, mRNA expressions of hyaluronan synthases and hyaluronidases were examined using real time RT-PCR. Inhibition of HA by 4-MU significantly decreased the migration, whereas promotion of HA by TPA or co-culture with tumor-derived fibroblasts significantly increased the migration of PDAC cells. The changes in HA production by these treatments tended to be associated with changes in HAS3 mRNA expression. Furthermore, addition of exogenous HA, especially low-molecular-weight HA, significantly increased the migration of PDAC cells. These findings suggest that HA stimulates PDAC cell migration and thus represents an ideal therapeutic target to prevent invasion and metastasis.
50Sodium hyaluronate supplemented culture medium combined with joint-simulating mechanical loading improves chondrogenic differentiation of human mesenchymal stem cells.PubMed
G Monaco, A J El Haj, M Alini, et al.
Eur Cell Mater. 2021 Jun 6;41:616-632. doi: 10.22203/eCM.v041a40.
In vitro models aim to recapitulate the in vivo situation. To more closely mimic the knee joint environment, current in vitro models need improvements to reflect the complexity of the native tissue. High molecular weight hyaluronan (hMwt HA) is one of the most abundant bioactive macromolecules in healthy synovial fluid, while shear and dynamic compression are two joint-relevant mechanical forces. The present study aimed at investigating the concomitant effect of joint-simulating mechanical loading (JSML) and hMwt HA-supplemented culture medium on the chondrogenic differentiation of primary human bone-marrow-derived mesenchymal stem cells (hBM-MSCs). hBM-MSC chondrogenesis was investigated over 28 d at the gene expression level and total DNA, sulphated glycosaminoglycan, TGF-β1 production and safranin O staining were evaluated. The concomitant effect of hMwt HA culture medium and JSML significantly increased cartilage-like matrix deposition and sulphated glycosaminoglycan synthesis, especially during early chondrogenesis. A stabilisation of the hBM-MSC-derived chondrocyte phenotype was observed through the reduced upregulation of the hypertrophic marker collagen X and an increase in the chondrogenic collagen type II/X ratio. A combination of JSML and hMwt HA medium better reflects the complexity of the in vivo synovial joint environment. Thus, JSML and hMwt HA medium will be two important features for joint-related culture models to more accurately predict the in vivo outcome, therefore reducing the need for animal studies. Reducing in vitro artefacts would enable a more reliable prescreening of potential cartilage repair therapies.
51Sodium Hyaluronate Supplemented Culture Media as a New hMSC Chondrogenic Differentiation Media-Model for Screening of Potential Cartilage Repair Therapies.PubMed
Graziana Monaco, Alicia Jennifer El Haj, Mauro Alini, et al.
Front Bioeng Biotechnol. 2020 Mar 31;8:243. doi: 10.3389/fbioe.2020.00243. eCollection 2020.
Surgical strategies to treat articular cartilage injury such as microfracture, expose human bone marrow stem cells (hMSCs) to synovial fluid and its components. High molecular weight hyaluronan (hMwt HA) is one of the most abundant bioactive macromolecules of healthy synovial fluid (hSF) and it plays an important role in the protection of opposing articular cartilage surfaces within the synovial joint. Although hMwt HA has been extensively used to attempt the engineering of the cartilage tissue, its effect as media supplement has not been established. Indeed, current media are often simple in their composition and doesn't recapitulate the rheological and biological features of hSF. In addition, critical molecules that can potentially change the chondrogenic behavior of hBMSCs to make the results more predictive of the real outcome, are lacking. In order to be one step closer to the physiology of hSF, a new culture media supplemented with physiological level of hMwt HA was developed and the effect of the hMwt HA on the chondrogenesis of hMSCs that would be present in a traumatic defect after marrow stimulation techniques, was investigated. hBMSC-seeded fibrin-polyurethane constructs were cultured in a serum free chondropermissive control medium (HA- TGFβ-). This medium was further supplemented with 10 ng/mL TGFβ1 (HA- TGFβ+) or 2 mg/ml hMwt HA 1.8 MDa (HA+ TGFβ-) or both (HA+ TGFβ+). Alternatively, 1 MDa HA was mixed with the fibrin at 0.2 mg/ml (HASc TGFβ+). The effect of hMwt HA on hMSC differentiation was investigated at the gene expression level by RT-qPCR and total DNA, sulfated glycosaminoglycans and Safranin O staining were evaluated. Addition of hMwt HA to the culture media, significantly increased the synthesis of sulfated glycosaminoglycans, especially in the early days of chondrogenesis, and reduced the upregulation of the hypertrophic cartilage marker collagen X. hMwt HA added inside the fibrin gel(HASc TGF+) led to the best matrix deposition. hMwt HA can be one key medium component in a more reliable system to reduce artifacts, enable more accurate pre-screening of potential cartilage repair therapies and reduce the need for animal studies.
52Effect of hyaluronic acid in bone formation and its applications in dentistry.PubMed
Ningbo Zhao, Xin Wang, Lei Qin, et al.
J Biomed Mater Res A. 2016 Jun;104(6):1560-9. doi: 10.1002/jbm.a.35681. Epub 2016 Apr 9.
Hyaluronic acid (HA), the simplest glycosaminoglycan, participates in several important biological procedures, including mediation of cellular signaling, regulation of cell adhesion and proliferation, and manipulation of cell differentiation. The effect of HA on cell proliferation and differentiation depends on its molecular weight (MW) and concentration. Moreover, the properties of high viscosity, elasticity, highly negative charge, biocompatibility, biodegradability, and nonimmunogenicity make HA attractive in tissue engineering and disease treatment. This review comprises an overview of the effect of HA on cell proliferation and differentiation in vitro, the role of HA in bone regeneration in vivo, and the clinical applications of HA in dentistry, focusing on the mechanism underlining the effect of MW and concentration of HA on cell proliferation and osteogenic differentiation. It is expected that practical progress of HA both in laboratory-based experiments and clinical applications will be achieved in the next few years. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1560-1569, 2016.
53Hyaluronic acid promotes angiogenesis by inducing RHAMM-TGFβ receptor interaction via CD44-PKCδ.PubMed
Deokbum Park, Youngmi Kim, Hyunah Kim, et al.
Mol Cells. 2012 Jun;33(6):563-74. doi: 10.1007/s10059-012-2294-1. Epub 2012 May 18.
Hyaluronic acid (HA) has been shown to promote angiogenesis. However, the mechanism behind this effect remains largely unknown. Therefore, in this study, the mechanism of HA-induced angiogenesis was examined. CD44 and PKCδ were shown to be necessary for induction of the receptor for HA-mediated cell motility (RHAMM), a HA-binding protein. RHAMM was necessary for HA-promoted cellular invasion and endothelial cell tube formation. Cytokine arrays showed that HA induced the expression of plasminogen activator-inhibitor-1 (PAI), a downstream target of TGFβ receptor signaling. The induction of PAI-1 was dependent on CD44 and PKCδ. HA also induced an interaction between RHAMM and TGFβ receptor I, and induction of PAI-1 was dependent on RHAMM and TGFβ receptor I. Histone deacetylase 3 (HDAC3), which is decreased by HA via rac1, reduced induction of plasminogen activator inhibitor-1 (PAI-1) by HA. ERK, which interacts with RHAMM, was necessary for induction of PAI-1 by HA. Snail, a downstream target of TGFβ signaling, was also necessary for induction of PAI-1. The down regulation of PAI-1 prevented HA from enhancing endothelial cell tube formation and from inducing expression of angiogenic factors, such as ICAM-1, VCAM-1 and MMP-2. HDAC3 also exerted reduced expression of MMP-2. In this study, we provide a novel mechanism of HA-promoted angiogenesis, which involved RHAMM-TGFβRI signaling necessary for induction of PAI-1.
54Dynamic Hyaluronan drives liver endothelial cells towards angiogenesis.PubMed
Sampa Ghose, Subhrajit Biswas, Kasturi Datta, et al.
BMC Cancer. 2018 Jun 11;18(1):648. doi: 10.1186/s12885-018-4532-1.
BACKGROUND: Angiogenesis, the formation of new blood vessels from pre-existing vasculature is essential in a number of physiological processes such as embryonic development, wound healing as well as pathological conditions like, tumor growth and metastasis. Hyaluronic acid (HA), a high molecular weight polysaccharide, major component of extracellular matrix is known to associate with malignant phenotypes in melanomas and various other carcinomas. Hyaluronic acid binding protein 1 (HABP1) has been previously reported to trigger enhanced cellular proliferation in human liver cancer cells upon its over-expression. In the present study, we have identified the HA mediated cellular behaviour of liver endothelial cells during angiogenesis. METHODS: Endothelial cells have been isolated from perfused liver of mice. Cell proliferation was studied using microwell plates with tetrazole dye. Cell migration was evaluated by measuring endothelial monolayer wound repair as well as through transwell migration assay. Alterations in proteins and mRNA expression were estimated by immunobloting and quantitative real time PCR using Applied Biosystems. The paraformaldehyde fixed endothelial cells were used for immuno- florescence staining and F-actin detection with conjugated antibodies. The images were captured by using Olympus florescence microscope (IX71). RESULTS: We observed that administration of HA enhanced cell proliferation, adhesion, tubular sprout formation as well as migration of liver endothelial cells (ECs). The effect of HA in the rearrangement of the actins confirmed HA -mediated cytoskeleton re-organization and cell migration. Further, we confirmed enhanced expression of angiogenic factors like VEGF-A and VEGFR1 in endothelial cells upon HA treatment. HA supplementation led to elevated expression of HABP1 in murine endothelial cells. It was interesting to note that, although protein levels of β- catenin remained unaltered, but translocation of this protein from membrane to nucleus was observed upon HA treatment, suggesting its role not only in vessel formation but also its involvement in angiogenesis signalling. CONCLUSIONS: The elucidation of molecular mechanism (s) responsible for HA mediated regulation of endothelial cells and angiogenesis contributes not only to our understanding the mechanism of disease progression but also offer new avenues for therapeutic intervention.
55A biphasic drug-releasing microneedle with ROS scavenging and angiogenesis for the treatment of diabetic ulcers.PubMed
Xinyue He, Lianghong Peng, Liming Zhou, et al.
Acta Biomater. 2024 Nov;189:270-285. doi: 10.1016/j.actbio.2024.09.045. Epub 2024 Oct 1.
Diabetic ulcers are one of the common complications in diabetic patients. Delayed wound healing is associated with persistent pro-inflammatory M1 polarization, reduced angiogenesis and increased reactive oxygen species (ROS) in the microenvironment. Wound healing consists of multiple phases and therefore requires treatment tailored to each phase. In this study, a biphasic drug-releasing microneedle (MN) was fabricated to achieve early ROS scavenging and late accelerated angiogenesis to promote wound healing. Vascular endothelial growth factor (VEGF) was first encapsulated in methacryloylated sulfonated chitosan (SCSMA) microspheres (V@MP), and then V@MP was loaded into hyaluronic acid (HA) microneedles along with cerium dioxide nanoparticles (CONPs). Rapid dissolution of HA rapidly releases the CONPs to clear ROS, whereas the V@MP stays in the wound. SCSMA slow degradation prolongs the release of VEGF, thereby promoting angiogenesis. In vitro and in vivo studies have shown that this biphasic drug-releasing smart microneedle improves cell proliferation and migration, effectively scavenges ROS, promotes angiogenesis and tissue regeneration, and synergistically promotes M2 macrophage polarization. It provides a new delivery mode for nano-enzymes and growth factors that could be multifunctional and synergistic in the treatment of diabetic ulcers. STATEMENT OF SIGNIFICANCE: In our study, we present a microneedle (V@MP/C@MN) that can release drugs biphasically, which showed good repair ability in diabetic ulcer model. Large amounts of CONPs were rapidly released to alleviate oxidative stress during the inflammation of the wound, and V@MP stayed in the wound for a long period of time to release VEGF and promote angiogenesis in the late stage of wound healing. The results indicated that V@MP/C@MN could promote cell proliferation and migration, effectively scavenge ROS, promote angiogenesis and tissue regeneration, and synergistically promote M2 macrophage polarization, which could play a multifunctional and synergistic role in the treatment of diabetic ulcers.
56Hyaluronic acid-based glucose-responsive antioxidant hydrogel platform for enhanced diabetic wound repair.PubMed
Zejun Xu, Guiting Liu, Ping Liu, et al.
Acta Biomater. 2022 Jul 15;147:147-157. doi: 10.1016/j.actbio.2022.05.047. Epub 2022 May 29.
Hyaluronic acid (HA)-based antioxidant hydrogels have achieved remarkable results in diabetic wound repair. However, the realization of their glucose-responsive antioxidant functions remains a significant challenge. In this study, we modified hyaluronic acid methacrylate (HAMA) with phenylboronic acid (PBA) and developed a glucose-responsive HA derivative (HAMA-PBA). A glucose-responsive HAMA-PBA/catechin (HMPC) hydrogel platform was then fabricated by forming a borate ester bond between HAMA-PBA and catechin. The results showed that the HMPC hybrid hydrogel not only had a three-dimensional network structure and Young's modulus similar to those of skin tissue, but also possessed biocompatibility. The HMPC hydrogel also showed unique glucose-responsive catechin release behavior and remarkable antioxidant capability, which could effectively eliminate intracellular reactive oxygen species and protect cells from oxidative stress damage (increased superoxide dismutase activity, stabilized reduced glutathione/oxidized glutathione ratio, and reduced malondialdehyde content). Additionally, in vitro and in vivo experimental results showed that the HMPC hydrogel effectively promoted angiogenesis (enhanced VEGF and CD31 expression) and reduced inflammatory responses (decreased IL-6 level and increased IL-10 level), thus rapidly repairing diabetic wounds (within three weeks). This was a significant improvement as compared to that observed for the untreated control group and the HMP hydrogel group. These results indicated the potential for the application of the HMPC hydrogel for treating diabetic wounds. STATEMENT OF SIGNIFICANCE: At present, the delayed closure rate of diabetic chronic wounds caused by excessive reactive oxygen species (ROS) remains a worldwide challenge. Hyaluronic acid (HA)-based antioxidant hydrogels have made remarkable achievements in diabetic wound repair; however, the realization of their glucose-responsive antioxidant functions is a tough challenge. In this work, we developed a novel HA-based hydrogel platform with glucose-responsive antioxidant activity for rapid repair of diabetic wounds. In vitro and in vivo experimental results showed that the HMPC hydrogel could effectively promote angiogenesis (enhanced VEGF and CD31 expression) and reduce inflammatory response (decreased IL-6 level and increased IL-10 level), thus rapidly repairing diabetic wounds (within 3 weeks). These results indicated the potential of the HMPC hydrogel for application in diabetic wound treatment.
57In vivo evaluation of MMP sensitive high-molecular weight HA-based hydrogels for bone tissue engineering.PubMed
Jungju Kim, In Sook Kim, Tae Hyung Cho, et al.
J Biomed Mater Res A. 2010 Dec 1;95(3):673-81. doi: 10.1002/jbm.a.32884.
Hyaluronic acid (170 kDa)-based hydrogel was synthesized using acrylated hyaluronic acid (HA) and matrix metalloproteinase (MMP) sensitive HA-based hydrogels were then prepared by conjugation with two different peptides: cell adhesion peptides containing integrin-binding domains (Arg-Gly-Asp: RGD) and a cross-linker with MMP degradable peptides to mimic the remodeling characteristics of natural extracellular matrices by cell-derived MMPs. Mechanical properties of these hydrogels were evaluated with different weight percentages (2.5 and 3.5 wt %) by measuring elastic modulus, viscous modulus, and swelling ratio. Human mesenchymal stem cells (hMSCs) were then cultured in MMP-sensitive or insensitive HA-based hydrogels and/or immobilized cell adhesive RGD peptides in vitro. Actin staining and image analysis proved that cells cultured in the MMP-sensitive hydrogel with RGD peptides showed extensive cell spreading and sprouting. Gene expression analysis showed that bone specific genes such as alkaline phosphatase, osteocalcin, and osteopontin increased in MMP-sensitive hydrogels as biomolecules such as BMPs and cells were added in the gels. For in vivo calvarial defect regeneration, five different samples (MMP insensitive hydrogel, MMP sensitive hydrogel, MMP sensitive hydrogel with BMP-2, MMP sensitive hydrogel with hMSC, and MMP sensitive hydrogel with BMP-2 and hMSC) were prepared. After 4 weeks of implantation, the Masson-Trichrome staining and micro computed tomography scan results demonstrated that the MMP sensitive hydrogels with BMP-2 and hMSCs have the highest mature bone formation. The MMP sensitive HA-based hydrogel could become useful scaffolds in bone tissue engineering with improvements on tissue remodeling rates and regeneration activity.
58Local delivery of hydrogel encapsulated vascular endothelial growth factor for the prevention of medication-related osteonecrosis of the jaw.PubMed
Dileep Sharma, Stephen Hamlet, Cedryck Vaquette, et al.
Sci Rep. 2021 Dec 3;11(1):23371. doi: 10.1038/s41598-021-02637-w.
The anti-angiogenic effects of bisphosphonates have been hypothesized as one of the major etiologic factors in the development of medication-related osteonecrosis of the jaw (MRONJ), a severe debilitating condition with limited treatment options. This study evaluated the potential of a gelatine-hyaluronic acid hydrogel loaded with the angiogenic growth factor, vascular endothelial growth factor (VEGF), as a local delivery system to aid in maintaining vascularization in a bisphosphonate-treated (Zoledronic Acid) rodent maxillary extraction defect. Healing was assessed four weeks after implantation of the VEGF-hydrogel into extraction sockets. Gross examination and histological assessment showed that total osteonecrosis and inflammatory infiltrate was significantly reduced in the presence of VEGF. Also, total vascularity and specifically neovascularization, was significantly improved in animals that received VEGF hydrogel. Gene expression of vascular, inflammatory and bone specific markers within the defect area were also significantly altered in the presence of VEGF. Furthermore, plasma cytokine levels were assessed to determine the systemic effect of locally delivered VEGF and showed similar outcomes. In conclusion, the use of locally delivered VEGF within healing extraction sockets assists bone healing and prevents MRONJ via a pro-angiogenic and immunomodulatory mechanism.
59In Vitro Hair Growth Promoting Effect of a Noncrosslinked Hyaluronic Acid in Human Dermal Papilla Cells.PubMed
Nicola Zerbinati, Sabrina Sommatis, Cristina Maccario, et al.
Biomed Res Int. 2021 Oct 31;2021:5598110. doi: 10.1155/2021/5598110. eCollection 2021.
Dermal papilla cells (DPCs) are a source of nutrients and growth factors, which support the proliferation and growth of keratinocytes as well as promoting the induction of new hair follicles and maintenance of hair growth. The protection from reactive oxygen species (ROS) and the promotion of angiogenesis are considered two of the basal mechanisms to preserve the growth of the hair follicle. In this study, a noncrosslinked hyaluronic acid (HA) filler (HYDRO DELUXE BIO, Matex Lab S.p.A.) containing several amino acids was tested with in vitro assays on human follicle dermal papilla cells (HFDPCs). The experiments were carried out to investigate the possible protection against oxidative stress and the ability to increase the vascular endothelial growth factor (VEGF) release. The results demonstrated the restoration of cell viability against UVB-induced cytotoxicity and an increase in the VEGF secretion. These data demonstrate the capability of the product to modulate human dermal papilla cells, suggesting a future use in mesotherapy, a minimally invasive local intradermal therapy (LIT), after further clinical investigations.
60MSC-derived exosomes injectable hyaluronic acid hydrogel for enhanced chronic wound healing.PubMed
Hanqing Yu, Jingyu Zhang, Liu Yang, et al.
J Control Release. 2025 Sep 10;385:113985. doi: 10.1016/j.jconrel.2025.113985. Epub 2025 Jun 26.
Diabetic chronic wounds are characterized by delayed healing and disrupted immune response. Mesenchymal stem cell-derived exosomes (MSC) hold significant potential for enhancing wound healing by facilitating intercellular communication, reducing excessive inflammation, and supporting cell proliferation. However, direct application of MSC to wound sites often results in rapid diffusion and poor retention, limiting their therapeutic efficacy. In this study, we developed a hyaluronic acid (HA)-based injectable hydrogel system to deliver MSC for treating diabetic chronic wounds. This hydrogel system exhibited excellent cytocompatibility, biodegradability, and skin-like rheology properties. The porous structure of the hydrogel system allows for in situ retention of exosomes, enabling sustained therapeutic effects on the wound. In vitro studies demonstrated that the hydrogels enhanced the proliferation and migration of endothelial cells and fibroblasts. In vivo studies confirmed the hydrogel's ability to accelerate wound closure, enhance angiogenesis, and promote re-epithelialization. This MSC loaded injectable hydrogel system provides sustained therapeutic benefits and enhances tissue regeneration, presenting a promising clinical strategy for the treatment of chronic diabetic wounds.
61pH- and glucose-responsive antioxidant hydrogel promotes diabetic wound healing.PubMed
Yanjun Zhang, Ling Zhang, Pingli Wu, et al.
Biomater Adv. 2025 Apr;169:214177. doi: 10.1016/j.bioadv.2025.214177. Epub 2025 Jan 9.
Excessive oxidative stress and persistent inflammation are key factors contributing to the formation of diabetic chronic wounds. Delivering antioxidants through a microenvironment-responsive hydrogel system can effectively enhance wound healing and tissue regeneration. In this study, we developed a novel pH- and glucose-responsive hydrogel using Schiff base reaction and phenyl borate group for intelligent antioxidant release. Hyaluronic acid (HA) modified with phenylboronic acid (PBA) (HA-PBA) was oxidized to form OHA-PBA, which was then crosslinked with carboxymethyl chitosan (CMCS) and incorporated Proanthocyanidins (PA) to create an OHA-PBA/CMCS/PA (OPCP) hydrogel. The reversible nature of imine and borate groups enabled the responsive release of PA from OPCP hydrogels under acidic and high glucose conditions. The OPCP hydrogel exhibited excellent biocompatibility, suitable mechanical properties, and biodegradability. Both in vitro and in vivo results demonstrated that the OPCP hydrogel effectively reduced reactive oxygen species (ROS), suppressed inflammation, promoted vascularization, accelerated collagen deposition, and facilitated diabetic wound healing. This strategy offers novel insights into microenvironment-responsive scaffolds, highlighting the potential application of this responsive antioxidant hydrogel scaffold for chronic diabetic wound treatment.
62Glucose and pH dual-responsive hydrogels with antibacterial, reactive oxygen species scavenging, and angiogenesis properties for promoting the healing of infected diabetic foot ulcers.PubMed
Zhendong Li, Longhui Chen, Shasha Yang, et al.
Acta Biomater. 2024 Dec;190:205-218. doi: 10.1016/j.actbio.2024.10.020. Epub 2024 Oct 16.
The healing process of diabetic foot ulcers is challenging due to the presence of a complex and severe inflammatory microenvironment, characterized by hyperglycemia, low pH, susceptibility to infection, vascular dysfunction, and over-expression of reactive oxygen species (ROS), which can potentially lead to amputation or even mortality. Herein, a glucose and pH dual-responsive hydrogel was designed and prepared by crosslinking phenylboronic acid-grafted quaternary chitosan (QF, 4 wt%) with dopamine-grafted oxidized hyaluronic acid (OD, 5 wt%) through phenylboronation, schiff-base reaction, and other techniques. The multifunctional QO/@PV@AB7 hydrogel was prepared by incorporating pravastatin-loaded chitosan nanoparticles (CSNPs@PV, 2 mg/mL) and antimicrobial peptide AMP-AB7 loaded silica nanoparticles (SiONPs@AB7, 0.5 mg/mL). The results demonstrate that the QO/@PV@AB7 hydrogel exhibits good responsiveness to acidic conditions and high glucose levels, while effectively scavenging various types of ROS. Moreover, it exerted protective effects against oxidative stress on cells, enhanced HUVECs viability, and promoted angiogenesis. Notably, the QO/@PV@AB7 hydrogel displayed potent antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli. Additionally, in an MRSA-infected rat model of diabetic foot wounds, administration of the QO/@PV@AB7 hydrogel led to increased secretion of pro-angiogenic factors such as vascular endothelial nitric oxide synthase (eNOS), vascular endothelial-generating factor (VEGF), and endothelial cell adhesion molecule (CD31). Furthermore, the hydrogel significantly reduced the levels of inflammatory factors such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), while simultaneously increasing the levels of anti-inflammatory cytokines such as interleukin-10 (IL-10). The findings suggest that multifunctional hydrogels incorporating PV@CSNPs and SiONPs@AB7 demonstrate promising potential as a therapeutic approach for the treatment of diabetic foot. STATEMENT OF SIGNIFICANCE: Here, a glucose and pH dual-responsive QO/@PV@AB7 hydrogel with antimicrobial and angiogenesis-promoting properties was developed for the treatment of infected wounds in diabetic feet. Our findings demonstrate that the proposed hydrogel exhibits good responsiveness, effectively scavenges various types of reactive oxygen species (DPPH, O, -OH, and ABTS+), provides protection against oxidative stress, enhances HUVECs cell viability, and promotes angiogenesis. Notably, it also demonstrates potent antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) and E. coli. Additionally, in vivo experiments demonstrated that the hydrogel exhibited accelerated wound healing in MRSA-infected diabetic foot ulcers, with a reduction of four days compared to the control group.
63Hyaluronic acid-Based wound dressings: A review.PubMed
Mariana F P Graça, Sónia P Miguel, Cátia S D Cabral, et al.
Carbohydr Polym. 2020 Aug 1;241:116364. doi: 10.1016/j.carbpol.2020.116364. Epub 2020 Apr 27.
Hyaluronic acid (HA), a non-sulfated glycosaminoglycan (GAG), is a major component of skin extracellular matrix (ECM) and it is involved in the inflammatory response, angiogenesis, and tissue regeneration process. Due to the intrinsic properties of HA (such as biocompatibility, biodegradability and hydrophilic character), it has been used to produce different wound dressings, namely sponges, films, hydrogels, and electrospun membranes. Herein, an overview of the different HA-based wound dressings that have been produced so far is provided as well as the future directions regarding the strategies aimed to improve the mechanical stability of HA-based wound dressings, along with the incorporation of biomolecules intended to ameliorate their biological performance during the healing process.
64High molecular weight hyaluronic acid improves salinity tolerance of Nile tilapia (Oreochromis niloticus) by mitigating apoptosis, alleviating oxidative stress and regulating carbohydrate metabolism.PubMed
Yuxi Yan, Jinquan Fan, Erchao Li, et al.
Int J Biol Macromol. 2025 Dec;332(Pt 2):148682. doi: 10.1016/j.ijbiomac.2025.148682. Epub 2025 Oct 31.
Salinity elevation induces hyperosmotic stress in fish, consistently triggering apoptosis. Apoptosis is recognized as the most sensitive indicator for assessing acute or severe salinity stress. Reducing the apoptosis under salinity stress may improve salinity tolerance of fish. Cells showed a significant decrease of hyaluronic acid content under hyperosmotic stress, suggesting that hyaluronic acid may play an important role in salinity adaptation. To investigate the functions and mechanisms by hyaluronic acid participating in osmoregulation, we explored in vitro and in vivo experiments separately. In vitro screening experiments, 100 μg/ml high molecular weight hyaluronic acid (HMW-HA, molecular weight = 1600 kDa) significantly improved cell survival rate under salinity stress. The HMW-HA scavenged ROS, enhanced cellular antioxidant capacity and inhibited apoptosis via the BCL2/caspase3 pathway. In the in vivo experiment, intraperitoneal injection of 90 mg/kg HMW-HA reduced the mortality rate of Nile tilapia (Oreochromis niloticus) under acute salinity stress. In addition to inhibiting apoptosis and improving antioxidant capacity, HMW-HA also enhanced glycolysis to produce more glucose, pyruvate and ATP for osmoregulation energy supplement Further, HMW-HA affected serum ion levels, while it had no significant effect on ion content in gills. HMW-HA alleviated gill damage such as gill filament disorder and inflammatory cell infiltration caused by high salinity. In summary, HMW-HA can enhance salinity tolerance of tilapia by inhibiting apoptosis, alleviating oxidative stress, and enhancing carbohydrate metabolism.
65Cytoprotective, anti-inflammatory, and antioxidant properties of high-molecular-weight hyaluronan enriched with red orange extract in human fibroblasts exposed to ultra violet light B irradiation.PubMed
Ilaria Penna, Ennio Albanesi, Rosalia Bertorelli, et al.
Biotechnol Appl Biochem. 2019 May;66(3):273-280. doi: 10.1002/bab.1722. Epub 2019 Jan 10.
Ultraviolet (UV) light exposure is the primary factor responsible for skin photoaging, affecting all the skin layers, mainly through the production of reactive oxygen species (ROS), activation of inflammatory responses, and apoptosis. In keeping with this evidence, exogenous supplementation with dietary antioxidants has been shown to provide photoprotective benefits. Moreover, oral administration of hyaluronic acid (HA) has been proved to reduce the signs of aged skin, such as wrinkles, and increase hydration and elasticity. The combination of different biologically active substances in order to slow down the onset of skin aging could represent a promising preventive strategy against photoaging. In the present study, we investigated the effects of a dietary supplement (IALUTEC RED), consisting of high-molecular-weight HA (HMW-HA) combined with red orange extract (ROC-Red Orange Complex ), in human fibroblasts exposed to ultra violet light B-induced oxidative stress. Our study suggests that, in fibroblasts exposed to UVB light, IALUTEC RED is active in decreasing both the inflammatory response and the generation of ROS, two events that are involved in skin photoaging.
66Antioxidant acitivity of low molecular weight hyaluronic acid.PubMed
Chunlin Ke, Lanping Sun, Deliang Qiao, et al.
Food Chem Toxicol. 2011 Oct;49(10):2670-5. doi: 10.1016/j.fct.2011.07.020. Epub 2011 Jul 20.
Two polysaccharides, low molecular weight hyaluronic acid-1 (LMWHA-1) and LMWHA-2, with their molecular weight of 1.45×10(5) and 4.52×10(4)Da, respectively, were prepared from high molecular weight hyaluronic acid (HA,1.05×10(6)Da). LMWHA-1, LMWHA-2 and HA were studied for their antioxidant activities. In vitro antioxidant assay, LMWHA showed strong inhibition of lipid peroxidation and scavenging activities of hydroxyl radical, moderate 1,1-diphenyl-2-picryldydrazyl radical and superoxide anion scavenging activity. In addition, the LMWHA-1 exhibited much stronger antioxidant activity than LMWHA-2 and HA. For antioxidant testing in vivo, LMWHA-1, LMWHA-2 and HA were orally administrated over a period of 7days in a cyclophosphamide(CY) induced immunosuppressed mice model. As results, administration of LMWHA was able to overcome CY-induced immunosuppression and significantly raised the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and total antioxidant capacity (TAOC) in immunosuppressed mice. The results showed that the LMWHA, possessing pronounced free radical scavenging and antioxidant activities.
67Antioxidant effect of hyaluronan on polymorphonuclear leukocyte-derived reactive oxygen species is dependent on its molecular weight and concentration and mainly involves the extracellular space.PubMed
Rafał Krasiński, Henryk Tchórzewski, Przemysław Lewkowicz
Postepy Hig Med Dosw (Online). 2009 May 4;63:205-12.
INTRODUCTION: Hyaluronan (HA), a component of the extracellular matrix, may regulate immune cell functions through its interactions with cellular receptors. Besides its effect on cytokine and chemokine production, its antioxidant properties have been described. However, the mechanisms of this are not fully elucidated. The aim of this study was to evaluate the relationship between HA concentration and molecular weight and its antioxidant properties towards human neutrophils. Also assessed was whether the antioxidant effect of HA is connected with a reduction in intracellular oxygen potential, which could indicate its direct effect on neutrophil respiratory burst. MATERIALS/METHODS: The relationship between HA's antioxidant properties and its concentration and molecular weight was assessed by the luminol-enhanced chemiluminescence method (CL). To evaluate the effect of HA on intracellular oxygen potential selectively, the dihydrorhodamine 123 (DHR123) flow cytometric method was used. RESULTS: Reduction of both HA molecular weight and its concentration decreased its antioxidant properties in the CL method. A selective effect of HA on intracellular oxygen potential measured by the DHR123 method was not shown. CONCLUSIONS: The antioxidant properties of HA are related to both its molecular weight and its concentration. The lack of an antioxidant effect of HA in the DHR123 test compared with a significant reduction in CL values at the same HA concentration suggests that HA acts mainly as a chemical ROI scavenger in the extracellular space.
68Saccharomyces cerevisiae fermentation of high molecular weight hyaluronic acid enhanced the antioxidant capacity in skin fibroblasts.PubMed
Meng Zhang, Doudou Shi, Mimi Cui, et al.
Arch Microbiol. 2025 Feb 20;207(3):66. doi: 10.1007/s00203-025-04274-7.
In this study, fermented hyaluronic acid (FHA) was obtained by fermentation of hyaluronic acid (HA) with molecular weight ranging from 1200 to 1300 kDa with two strains of Saccharomyces cerevicae (CICC 1308 and CICC 1747). Active substance content and spectral analysis revealed a substantial decrease in FHA molecular weight, accompanied by changes in monosaccharide composition, yet the chemical structure of FHA remained unchanged. Secondly, FHA exhibited a strong ability to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radicals. In hydrogen peroxide (HO)-induced human skin fibroblasts (HSF), FHA enhanced superoxide dismutase (SOD) and catalase (CAT) activities and reduced malondialdehyde (MDA) levels, reversing oxidative stress via the Nrf2/Keap1/HO-1 signaling pathway. These findings highlight FHA's potential as a natural cosmetic ingredient. Future research should investigate its long-term effects and potential applications in skin care treatments.
69Glycosaminoglycan and proteoglycan in skin aging.PubMed
Dong Hun Lee, Jang-Hee Oh, Jin Ho Chung
J Dermatol Sci. 2016 Sep;83(3):174-81. doi: 10.1016/j.jdermsci.2016.05.016. Epub 2016 May 27.
Glycosaminoglycans (GAGs) and proteoglycans (PGs) are abundant structural components of the extracellular matrix in addition to collagen fibers. Hyaluronic acid (HA), one of GAGs, forms proteoglycan aggregates, which are large complexes of HA and HA-binding PGs. Their crosslinking to other matrix proteins such as the collagen network results in the formation of supermolecular structures and functions to increase tissue stiffness. Skin aging can be classified as intrinsic aging and photoaging based on the phenotypes and putative mechanism. While intrinsic aging is characterized by a thinned epidermis and fine wrinkles caused by advancing age, photoaging is characterized by deep wrinkles, skin laxity, telangiectasias, and appearance of lentigines and is mainly caused by chronic sun exposure. The major molecular mechanism governing skin aging processes has been attributed to the loss of mature collagen and increased matrix metalloproteinase expression. However, various strategies focusing on collagen turnover remain unsatisfactory for the reversal or prevention of skin aging. Although the expression of GAGs and PGs in the skin and their regulatory mechanisms are not fully understood, we and others have elucidated various changes in GAGs and PGs in aged skin, suggesting that these molecules are important contributors to skin aging. In this review, we focus on skin-abundant GAGs and PGs and their changes in human skin during the skin aging process.
70Trends in polysaccharide-based hydrogels for skin anti-aging and skin antioxidant.PubMed
Kit-Leong Cheong, Qi Chen, Jude Juventus Aweya, et al.
Int J Biol Macromol. 2025 Aug;319(Pt 2):145366. doi: 10.1016/j.ijbiomac.2025.145366. Epub 2025 Jun 19.
Skin aging is a complex biological process influenced by both intrinsic and extrinsic factors, with oxidative stress playing a central role in accelerating dermal damage, pigmentation, inflammation, and mitochondrial dysfunction. In recent years, polysaccharide-based hydrogels have emerged as promising materials for anti-aging and antioxidant skincare due to their excellent biocompatibility, moisture retention, and bioactivity. This review highlights the mechanisms of skin aging, the pathological role of reactive oxygen species (ROS), and the need for effective, natural interventions. We explore diverse polysaccharides-such as alginate, carrageenan, agarose, cellulose, chitosan, β-glucan, hyaluronic acid, pectin, and starch-and their unique contributions to hydrogel performance in skin applications. Crosslinking strategies, including physical, chemical, enzymatic, click chemistry, and 3D printing, are discussed for tailoring hydrogel structure and function. We also examine the underlying mechanisms through which these hydrogels act: enhancing skin hydration, scavenging ROS, delivering antioxidants, reducing inflammation, and protecting against UV-induced photoaging. Together, these systems offer a synergistic approach to maintaining skin health and reversing age-related deterioration. With continued innovation in material design and delivery technologies, polysaccharide-based hydrogels represent a sustainable and powerful platform for next-generation anti-aging skincare solutions.
71Hyaluronic acid modified chuanxiong oil liposomes as a novel therapeutic agent for photoaging prevention.PubMed
Xiaofang He, Yue Hu, Yuanyuan Wu, et al.
Sci Rep. 2025 Apr 10;15(1):12237. doi: 10.1038/s41598-025-97450-0.
The gradual increase in ultraviolet B (UVB) health hazards to human skin, coupled with the irritation associated with existing sunscreen products, underscores the critical need for the development of natural sunscreens to combat UVB-induced photoaging. Chuanxiong oil (CXO) and hyaluronic acid (HA) possess excellent antioxidant and anti-apoptotic properties, which are closely linked to the mechanisms of photoaging. In this study, a composite nano-system (HA-CXO-Lip) comprising chuanxiong oil (CXO) and hyaluronic acid (HA) was initially fabricated. Subsequently, both in vitro HaCaT cell models and in vivo murine photoaging models were established to systematically evaluate the therapeutic efficacy and mechanistic actions of HA-CXO-Lip against photoaging under controlled experimental conditions. The investigation encompassed comprehensive assessments of its pharmacological effects and underlying molecular mechanisms through multimodal experimental approaches. Vitro experiments showed HA-CXO-Lip significantly reduced intracellular reactive oxygen species (ROS) levels and senescence-associated β-galactosidase (SA-β-Gal) activity. Furthermore, HA-CXO-Lip restored the levels of antioxidant enzymes, including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and hydroxyproline (HYP), while also decreasing the levels of lipid metabolites such as 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA). These findings indicate that HA-CXO-Lip effectively inhibits excessive oxidative stress. Additionally, HA-CXO-Lip inhibited apoptosis by reducing Bax levels and enhancing Bcl-2 expression in HaCaT cells. In vivo studies demonstrated that HA-CXO-Lip significantly reduced UVB irradiation-induced erythema and epidermal thickening in the backs of mice. It restored the orderly arrangement of collagen fibers and inhibited the activation of the core senescence pathway, AKT/mTOR, along with the downstream expression of matrix metalloproteinase 9 (MMP9), resulting in a decrease in collagen I disassembly. Additionally, HA-CXO-Lip was shown to significantly decrease the number of apoptotic cells, as indicated by the expression of the apoptosis marker cleaved cysteine aspartic protease-3 (C-Caspase-3) and the surface type I transmembrane glycoprotein (CD44), thereby further inhibiting apoptosis. The findings of this study suggest that HA-CXO-Lip can exert anti-photoaging effects through its antioxidant and anti-apoptotic properties, highlighting the synergistic efficacy of CXO and HA, which holds promise for the prevention and treatment of photoaging.
72β-Nicotinamide Mononucleotide Enhances Skin Barrier Function and Attenuates UV-B-Induced Photoaging in Mice.PubMed
Sung Jin Kim, Sullim Lee, Yea Jung Choi, et al.
Antioxidants (Basel). 2025 Nov 27;14(12):1424. doi: 10.3390/antiox14121424.
Ultraviolet B (UV-B) radiation significantly contributes to skin photoaging, which is characterized by epidermal thickening, collagen degradation, wrinkle formation, barrier dysfunction, and oxidative stress. Nicotinamide mononucleotide (NMN), a key precursor of nicotinamide adenine dinucleotide, regulates cellular energy metabolism and antioxidant defense and demonstrates anti-aging effects in animal models. Here, we investigated the protective effects of oral NMN supplementation against UV-B-induced photoaging in SKH-1 hairless mice. Over a 10-week experimental period, oral NMN administration significantly alleviated epidermal hypertrophy, reduced wrinkle formation and skin surface roughness, improved hydration and elasticity, and restored transepidermal water loss to near-normal levels. Histological analyses revealed marked preservation of collagen fiber density and attenuation of dermal matrix degradation. Furthermore, NMN supplementation inhibited the phosphorylation of MAPK signaling components (ERK, JNK, and p38), suppressed pro-inflammatory cytokine (TNF-α and IL-6) and matrix-degrading enzyme (MMP-1) expression, and restored hyaluronan synthase (HAS-1 and HAS-2) expression. Additionally, NMN enhanced the systemic antioxidant defense, as indicated by the restored superoxide dismutase activity. Thus, NMN has multi-layered protective effects against UV-B-induced skin aging by modulating oxidative stress, inflammatory signaling, extracellular matrix remodeling, and hyaluronic acid metabolism.
73Hyaluronic acid and HA-modified cationic liposomes for promoting skin penetration and retention.PubMed
Chang Ni, Zijun Zhang, Yuling Wang, et al.
J Control Release. 2023 May;357:432-443. doi: 10.1016/j.jconrel.2023.03.049. Epub 2023 Apr 18.
Hyaluronic acid (HA) has been widely used in cosmetics and topical preparations owing to its favorable moisturizing property and potential in enhancing drugs' skin permeability. Here, the influencing factors and underlying mechanism of HA on skin penetration were carefully investigated, and HA-modified Undecylenoyl-Phenylalanine (UP) liposomes (HA-UP-LPs) were designed as a proof of principle for efficacious transdermal drug delivery strategy to enhance the skin penetration and retention. An in vitro penetration test (IVPT) of HA with different molecular weights showed that low molecular weight HA (LMW-HA, 5 kDa and 8 kDa) could pass through the stratum corneum (SC) barrier and enter into the epidermis and dermis layers, whereas its high molecular counterparts (HMW-HA) were trapped on the SC surface. Mechanistic studies revealed that LMW-HA could interact with keratin and lipid in the SC meanwhile exerted a substantial skin hydration effect, which may partially contribute to the SC penetration benefit. In addition, the surface decoration of HA drove an energy-dependent caveolae/lipid raft-mediated endocytosis of the liposomes through direct binding to the CD44 receptors widely expressed on skin cell membranes. Notably, IVPT showed a 1.36-fold and 4.86-fold increase in skin retention of UP and a 1.62-fold and 5.41-fold increase in skin penetration of UP with HA-UP-LPs over UP-LPs and free UP at 24 h, respectively. As a result, the anionic HA-UP-LPs (-30.0 mV) showed enhanced drug skin penetration and retention compared with conventional cationic bared UP-LPs (+21.3 mV) on both in vitro mini-pig skin as well as in vivo mouse skin. Overall, the usage of LMW-HA might offer opportunities in developing novel topical preparations and skin care products with improved transdermal penetration and retention.
74Efficacy of cream-based novel formulations of hyaluronic acid of different molecular weights in anti-wrinkle treatment.PubMed
Tatjana Pavicic, Gerd G Gauglitz, Peter Lersch, et al.
J Drugs Dermatol. 2011 Sep;10(9):990-1000.
BACKGROUND: Due to its strong water-binding potential, hyaluronic acid (HA) is a well-known active ingredient for cosmetic applications. Native HA is proposed to help the skin to retain and maintain elasticity, turgor and moisture. OBJECTIVE: To observe the efficacy of topical application of 0.1% hyaluronan formulations of different molecular weights (MW) (50, 130, 300, 800 and 2000 kDa, respectively) in the periocular area as anti-wrinkle treatment. MATERIAL AND METHODS: Seventy-six female subjects between 30 and 60 years of age with clinical signs of periocular wrinkles applied one of the formulations twice-daily to the area of interest in a randomized fashion for 60 days. Around the other eye, a vehicle control cream was applied. Measurements of skin hydration and skin elasticity were performed before treatment, 30 and 60 days thereafter. At similar time points negative replicas were taken and evaluated by semi-automated morphometry. RESULTS: All HA-based creams utilized in this study demonstrated a significant improvement in skin hydration and overall elasticity values (R2) when compared to placebo. Measurements of wrinkle depth using mean roughness (Ra) and maximum roughness (Rz) values revealed significant improvement in the 130 and the 50 kDa HA group after 60 days of treatment compared to placebo-treated area. CONCLUSION: Topical application of all 0.1% HA formulations used in this study led to significant improvement in skin hydration and elasticity. Application of low-molecular-weight (LMW) HA was associated with significant reduction of wrinkle depth, which may be due to better penetration abilities of LMW HA.
75Benefits of topical hyaluronic acid for skin quality and signs of skin aging: From literature review to clinical evidence.PubMed
Bruna Bravo, Priscila Correia, José Euzébio Gonçalves Junior, et al.
Dermatol Ther. 2022 Dec;35(12):e15903. doi: 10.1111/dth.15903. Epub 2022 Oct 21.
Skin aging goes beyond a chronological process and also results from extrinsic factors referred to as the exposome. Hyaluronic acid (HA) is an important component of the extracellular matrix, with loss starting at 25 years old. While many studies of HA concern topical use, few literature reviews only address the use of topical HA in dermatology. This review describes the different characteristics of HA-containing cosmeceuticals, with a focus on skin aging and the impact of exposome factors on HA synthesis and degradation. A review was performed using the terms HA, hyaluronan, topical, dermatology, cosmetic, aging treatment, exposome, and cosmeceuticals. Results are also presented from a recent randomized controlled trial (RCT), which investigated the additional benefit of using a HA epidermic filler (HA-filler serum) combined with Botulinum toxin type A (BoNTA) to treat signs of skin aging. Subjects were randomized to two groups: HA-filler serum starting 24 h after the BoNTA injection then twice daily for 24 weeks, or the control group, which received BoNTA. HA is a key ingredient used in cosmeceuticals for its hydration/antiaging properties (hygroscopic, rheological, and viscoelastic). Several clinical studies indicate that HA is both well tolerated and effective, adjuvant to both post-surgical and facial rejuvenation procedures. In the RCT, one of few studies to combine BoNTA and HA with a 6-month follow-up, the HA-filler serum lengthened the duration of BoNTA's effect in reducing wrinkles. Numerous studies support HA-based cosmeceuticals as a noninvasive, effective solution for improving skin hydration and rejuvenation.
76Hyaluronic Acid Basics and Rheology.PubMed
Grace T Wu, Joanna Kam, Jason D Bloom
Clin Plast Surg. 2023 Jul;50(3):391-398. doi: 10.1016/j.cps.2022.12.004.
Hyaluronic acid (HA) is the most common dermal filler in use. It improves wrinkles and volume loss not only by filling and volumizing but also by hydrating the injected area with its water affinity. It is a naturally occurring component of skin, and there is a negligible risk of immunologic or allergic reaction with injection. It is rapidly degraded by the injection of hyaluronidase, thus creating an ideal injectable material that is low risk and reversible. Its duration of effect may be longer than expected based on bioavailability of the HA product due to collagen synthesis or fibroblast stimulation.
77Looking older: fibroblast collapse and therapeutic implications.PubMed
Gary J Fisher, James Varani, John J Voorhees
Arch Dermatol. 2008 May;144(5):666-72. doi: 10.1001/archderm.144.5.666.
Skin appearance is a primary indicator of age. During the last decade, substantial progress has been made toward understanding underlying mechanisms of human skin aging. This understanding provides the basis for current use and new development of antiaging treatments. Our objective is to review present state-of-the-art knowledge pertaining to mechanisms involved in skin aging, with specific focus on the dermal collagen matrix. A major feature of aged skin is fragmentation of the dermal collagen matrix. Fragmentation results from actions of specific enzymes (matrix metalloproteinases) and impairs the structural integrity of the dermis. Fibroblasts that produce and organize the collagen matrix cannot attach to fragmented collagen. Loss of attachment prevents fibroblasts from receiving mechanical information from their support, and they collapse. Stretch is critical for normal balanced production of collagen and collagen-degrading enzymes. In aged skin, collapsed fibroblasts produce low levels of collagen and high levels of collagen-degrading enzymes. This imbalance advances the aging process in a self-perpetuating, never-ending deleterious cycle. Clinically proven antiaging treatments such as topical retinoic acid, carbon dioxide laser resurfacing, and intradermal injection of cross-linked hyaluronic acid stimulate production of new, undamaged collagen. Attachment of fibroblasts to this new collagen allows stretch, which in turn balances collagen production and degradation and thereby slows the aging process. Collagen fragmentation is responsible for loss of structural integrity and impairment of fibroblast function in aged human skin. Treatments that stimulate production of new, nonfragmented collagen should provide substantial improvement to the appearance and health of aged skin.
78A hyaluronic acid-based micro-filler improves superficial wrinkles and skin quality: a randomized prospective controlled multicenter study.PubMed
Ferial Fanian, Jean-Jacques Deutsch, Marie Thérèse Bousquet, et al.
J Dermatolog Treat. 2023 Dec;34(1):2216323. doi: 10.1080/09546634.2023.2216323. Epub 2023 Aug 14.
BACKGROUND: NCTF®135HA is a bio-revitalizing solution containing hyaluronic acid designed to compensate for skin dehydration, fatigue, and fine wrinkles associated with endogenous and environmental aging. METHODS: We conducted a randomized, active-controlled study to evaluate the efficacy and tolerability of NCTF®135HA injections on the face (crow's feet), neck, and décolleté regions. Subjects were randomly assigned (3:1) to receive three NCTF®135HA treatment sessions plus twice-daily anti-aging moisturizer cream or cream alone (control). The primary outcome was the reduction in superficial wrinkles between baseline and Day (D)75 in the three areas, assessed by profilometric measures, clinical scoring, subjective changes, and tolerability. RESULTS: 146 subjects were randomized to NCTF®135HA ( = 107) or control ( = 38). At D75 and D120, NCTF®135HA significantly reduced wrinkles in all three areas and improved facial radiance scores compared with the control. Skin hydration significantly increased 7 d after the last NCTF®135HA injection. Self-esteem scales showed statistically significant improvements at D75 and D120 in subjects treated with NCTF®135HA versus baseline. Most adverse events were mild, resolved within 48 h, and were related to the injection procedure. CONCLUSION: NCTF®135HA is an effective and well-tolerated treatment to reduce the skin signs of aging. The results are significantly superior to a routine anti-aging cream alone.(Funded by Laboratories FILLMED; ID-RCB number: 2018-A03167-48; clinicaltrials.gov number: NCT05609617).
79Hyaluronic acid and wound healing.PubMed
Manuela G Neuman, Radu M Nanau, Loida Oruña-Sanchez, et al.
J Pharm Pharm Sci. 2015;18(1):53-60. doi: 10.18433/j3k89d.
BACKGROUND: We developed an experimental model of ethanol-induced dermatotoxicity and hepatocytoxicity using normal human keratinocytes and normal human hepatocytes that preserve inducible cytochrome p450 activities. The original work was described in several articles. The objective of this study was to determine whether hyaluronic acid attenuates skin necrosis, and to further clarify its uses in wound repair in humans, animal models and in vitro studies. METHODS: We performed a systematic review of the literature using the terms "hyaluronic acid" and "wound healing". PubMed was searched for studies published during the period 2010-2014. RESULTS: Hyaluronic acid is used in tissue regeneration alone or in combination with herbal or Western medicine. Scaffolds made up of hyaluronic acid were used to embed basic fibroblast growth factor. CONCLUSION: Hyaluronic acid extracts are safe and efficacious products to be used in skin repair.
80Wound healing through synergy of hyaluronan and an iodine complex.PubMed
K F Cutting
J Wound Care. 2011 Sep;20(9):424, 426, 428-30. doi: 10.12968/jowc.2011.20.9.424.
Hyaluronan, a glycosaminoglycan (GAG), is a polysaccharide found in many locations in the human body, such as eye, skin and soft tissue. It is also found in other mammals and bacteria. As a component of the extracellular matrix, its role in wound repair, among others, is that of providing a temporary structure to support new tissue formation. Harnessing the therapeutic action of hyaluronan into a topical application of proven clinical benefit has proved challenging. A new development in hyaluronan technology, comprising sodium hyaluronate and iodine complex, offers a novel approach in exploiting the benefits of hyaluronan and delivering real clinical benefits for a wide range of wound types.
81The Effects of Dietary Supplementation with Collagen and Vitamin C and Their Combination with Hyaluronic Acid on Skin Density, Texture and Other Parameters: A Randomised, Double-Blind, Placebo-Controlled Trial.PubMed
Katja Žmitek, Janko Žmitek, Hristo Hristov, et al.
Nutrients. 2024 Jun 17;16(12):1908. doi: 10.3390/nu16121908.
Collagen dietary supplements are becoming increasingly popular as a means to reduce signs of skin ageing. The objective of this three-way, randomised, placebo-controlled, double-blind study was to examine and contrast the effects of dietary supplementation with a daily dose of 5 g hydrolysed collagen with 80 mg of vitamin C (CP product) and their combination with 30 mg of hyaluronic acid (CPHA product) over 16 weeks. Validated methods were utilised for the objective evaluation of skin parameters. In total, 87 subjects (women, 40-65 years) completed the entire trial, distributed across the groups as follows: placebo group ( = 29), CPHA group ( = 28), and CP group ( = 30). The results showed beneficial effects of both test products, with notable enhancements in dermis density, skin texture, and a reduction in the severity of wrinkles. In contrast, the administration of either of the products did not yield any significant impacts on skin elasticity or hydration. Observation of the investigated skin parameters did not show superior effects of the addition of hyaluronic acid (HA) to collagen. Therefore, the ability of supplementation with HA to improve the effects on investigated skin parameters beyond the supplementation of collagen alone cannot be confirmed.
82Methicillin-resistant Staphylococcus aureus colonization of infectious and non-infectious skin and soft tissue lesions in patients in Tehran.PubMed
Haniyeh Khalili, Shahin Najar-Peerayeh, Mona Mahrooghi, et al.
BMC Microbiol. 2021 Oct 18;21(1):282. doi: 10.1186/s12866-021-02340-w.
BACKGROUND: The most common clinical manifestations of Staphylococcus aureus strains in the community are skin and soft-tissue infections. S. aureus could colonize the body sites and complicate the pathogenesis of skin diseases. S. aureus colonization is a risk factor for severe conditions such as bone and joint infections, pneumonia, bacteremia, and endocarditis. This study aimed to investigate the prevalence of S. aureus strains in skin and soft tissue infections and other skin disorders in patients referring to dermatology clinics and to evaluate the antibiotic resistance pattern and molecular characteristics of S. aureus isolates. METHODS: Skin swabs were collected from the lesional sites in 234 outpatients referring to dermatology clinics in three hospitals in Tehran. Antibiotic susceptibility, biofilm formation, and hemolysis tests were performed for isolates. PCR was done for SCCmec typing, agr grouping, and virulence genes detecting. RESULTS: The prevalence of S. aureus strains among patients with skin and soft-tissue infections and other skin lesions was 44.77% (30/67) and 44.91% (75/167), respectively. Also, 59 (56.19%) isolates were MRSA, 35.57% were HA-MRSA, and 30.5% were CA-MRSA. The psmα gene was more prevalent (62.8%) among isolates, followed by hlaα (56.1%), tsst-1 (15.2%) eta (13.3%), etb (6.6%), and pvl (2.8%). The agr specificity groups I, II, III, and IV were identified in 49.5, 21.9, 11.4, and 14.2% of S. aureus isolates, respectively. Most (56%) S. aureus isolates produced a moderate biofilm, and 23.8% of them produced strong biofilms. α-hemolysin (46.6%), β-hemolysin (25.7%), γ-hemolysin (19%), and both α and β-hemolysin (5.7%) were also produced by isolates. CONCLUSION: The present study results indicated high colonization of skin lesions by HA-MRSA and CA-MRSA clones; MRSA strains were more resistant to antibiotics, contained various toxin genes, and were able to form biofilms. Therefore, they could play a vital role in the pathogenesis of various skin diseases; also, they could spread and cause infections in other body sites. Eradication and decolonization strategies could prevent recurrent infections and the spread of resistant strains and improve skin conditions.
83Preparation, characterization, and Staphylococcus aureus biofilm elimination effect of baicalein-loaded tyrosine/hyaluronic acid/β-cyclodextrin-grafted chitosan nano-delivery system.PubMed
Ling Tang, Zhongbin Zhang, Wenyou Ding, et al.
Int J Biol Macromol. 2024 Jan;254(Pt 3):128066. doi: 10.1016/j.ijbiomac.2023.128066. Epub 2023 Nov 12.
Staphylococcus aureus (S. aureus) is an important cause of infections associated with implanted medical devices due to the formation of bacterial biofilm, which can prevent the penetration of drugs, thus posing a serious multi-drug resistance. Methicillin-resistant Staphylococcus aureus (MRSA) is one of them. In order to enhance the biofilm elimination effect of Baicalein (BA), a BA-loaded Tyr/HA/CD-CS nano-delivery system was successfully prepared using β-cyclodextrin grafted with chitosan (CD-CS), Hyaluronic Acid (HA), and D-Tyrosine (D-Tyr). The Tyr/HA/CD-CS-BA-NPs have a uniform particle size distribution with a particle size of 238.1 ± 3.06 nm and a PDI of 0.130 ± 0.02. The NPs showed an obvious inhibitory effect on planktonic bacteria with a MIC of 12.5 μg/mL. In vivo and in vitro tests showed that the NPs could enhance the elimination effect of BA on the MRSA biofilm. The results of Confocal Laser Scanning Microscopy (CLSM), Live & Dead Kit, and colony count experiments illustrated that Tyr/HA/CD-CS-BA-NPs could enhance the permeability of drugs to the biofilm and improve the ability to kill the biofilm bacteria, which may be an important mechanism to enhance the elimination of the MRSA biofilm. These findings will help develop new, effective medicaments for treating bacterial biofilm infections.
84Methacrylated sericin/methacrylated hyaluronic acid composite microneedles with exosomes for diabetic wound healing.PubMed
Xinhao Xu, Yuxin Shen, Jun Wu, et al.
Int J Biol Macromol. 2025 Jul;318(Pt 1):144957. doi: 10.1016/j.ijbiomac.2025.144957. Epub 2025 Jun 3.
Skin wounds that arise from diabetes and chronic wound conditions present significant challenges in clinical care. Microneedle patches can puncture the skin epidermis and deliver drugs directly to the injured tissue rendering them particularly effective for wound healing, while hydrogel offers a physical barrier, moisture retention, and oxygen permeability at wound sites. Therefore, in this study, a novel microneedle patch drug delivery system that employs methacrylated sericin and methacrylated hyaluronic acid as substrates was developed. Exosomes released from the tip portion are found to improve the proangiogenic, and reactive oxygen species and inflammation inhibitory properties of the microneedle patch. A zeolite imidazole framework-8 encapsulated in the backing portion of the microneedle to form a protective layer over the wound surface inhibits bacteria and prevents infection by foreign pathogens. The designed multifunctional microneedle patch accelerates the healing of diabetic wounds, offering a promising therapeutic approach.
85A bimetallic catalyst core-shell nanoplatform enabling all-stage management of infected diabetic wound healing.PubMed
Huihui Xie, Lan Zhang, Tiexin Ding, et al.
Biomaterials. 2026 Apr;327:123771. doi: 10.1016/j.biomaterials.2025.123771. Epub 2025 Oct 8.
The impaired healing of diabetic wounds is largely attributable to the pathological microenvironment, which promotes susceptibility to bacterial infection, hyperinflammation, vasculopathy, and neuropathy. To address these issues, a bimetallic catalyst core-shell nanoplatform ((Cu@Ce)HM) with Cu-bonded Ce-MOF as core and HHC36-loaded hyaluronic acid (HA) as shell is constructed. (Cu@Ce)HM demonstrates potent antibacterial efficacy (eliminating 97.01 % of Staphylococcus aureus (S. aureus) and 99.55 % of Escherichia coli (E. coli)) through the responsive release of Cu ions and HHC36 in infected diabetic microenvironment. Furthermore, (Cu@Ce)HM effectively scavenges excessive reactive oxygen species (ROS) via its superoxide dismutase (SOD)- and catalase (CAT)-mimicking enzymatic activities. Density functional theory (DFT) calculations reveals that Cu bonding induces electron rearrangement within the Ce-MOF, stabilizing the coexistence of Cu/Cu and Ce/Ce redox couples. This bimetallic synergy enables efficient catalysis, improving ROS scavenging performance of (Cu@Ce)HM. (Cu@Ce)HM improves the pathological microenvironment by scavenging ROS and supplying low-dose Cu ions, and thus promotes the recovery and M2 phenotype of M1 macrophages, enhancing anti-inflammatory cytokine secretion. This immunomodulation further augments the biofunctions of endothelial cells (HUVECs) and neural cells (PC12). Transcriptome sequencing analysis indicates that the antioxidant properties and Cu supplementation provided by (Cu@Ce)HM promotes the recovery and differentiation of damaged PC12 cells by upregulating key signaling pathways, including PI3K/Akt/CREB, Ras/ERK/CREB, and Ras/ERK/MAPK. This study presents a promising "all-stage" management strategy for infected diabetic wound regeneration, integrating infection elimination, immunoregulation, and the facilitation of neuroangiogenesis and extracellular matrix (ECM) remodeling.
86Novel peptide and hyaluronic acid coated biomimetic liposomes for targeting bacterial infections and sepsis.PubMed
Eman A Ismail, Calvin A Omolo, Mohammed A Gafar, et al.
Int J Pharm. 2024 Sep 5;662:124493. doi: 10.1016/j.ijpharm.2024.124493. Epub 2024 Jul 22.
Sepsis is a life-threatening syndrome resulting from an imbalanced immune response to severe infections. Despite advances in nanomedicines, effective treatments for sepsis are still lacking. Herein, vancomycin free base (VCM)-loaded dual functionalized biomimetic liposomes based on a novel TLR4-targeting peptide (P3) and hyaluronic acid (HA) (HA-P3-Lipo) were developed to enhance sepsis therapy. The nanocarrier revealed appropriate physicochemical parameters, good stability, and biocompatibility. The release of VCM from HA-P3-Lipo was found to be sustained with 76 % VCM released in 48 h. The biomimicry was elucidated by in silico tools and MST and results confirmed strong binding between the system and TLR4. Furthermore, HA-P3-Lipo revealed 2-fold enhanced antibacterial activity against S. aureus, sustained antibacterial activity against MRSA over 72 h and 5-fold better MRSA biofilm inhibition compared to bare VCM. Bacterial-killing kinetics and flow cytometry confirmed the superiority of HA-P3-Lipo in eliminating MRSA faster than VCM. The in vivo potential of the nanocarrier was elucidated in an MRSA-induced sepsis mice model, and the results confirmed the superiority of HA-P3-Lipo compared to free VCM in eliminating bacteria and down-regulating the proinflammatory markers. Therefore, HA-P3-Lipo exhibits potential as a promising novel multi-functional nanosystem against sepsis and could significantly contribute to the transformation of sepsis therapy.
87Multifunctional hyaluronic acid microneedle patch enhances diabetic wound healing in diabetic infections.PubMed
Quanyi Lu, Xiyue Tang, Bailong Tao, et al.
Int J Biol Macromol. 2025 Mar;296:139685. doi: 10.1016/j.ijbiomac.2025.139685. Epub 2025 Jan 10.
Diabetic wounds often exhibit a chronic non-healing state due to the combined effects of multiple factors, including hyperglycemia, impaired angiogenesis, immune dysfunction, bacterial infection, and excessive oxidative stress. Despite the availability of various therapeutic strategies, effectively managing the complex and prolonged healing process of diabetic infected wounds remains challenging. In this study, we combined the natural antidiabetic drug lipoic acid (LA) with the RADA16-YIGSR (RY) peptide obtained through solid-phase synthesis, utilizing reversible hydrogen bonds and coordination bonds for binding. Driven by strong electrostatic interactions and the spontaneous hydrophobic aggregation of LA molecules, the peptide underwent a self-assembly process without requiring additional external stimuli. Subsequently, this self-assembled peptide was loaded into a microneedle (MN) system composed of hyaluronic acid (HA).HA(RY/LA) MNs utilize LA's sustained release to effectively eliminate S. aureus and E. coli, reduce ROS, promote M2 macrophage polarization via NF-κB, and induce neovascularization through synergistic LA and RY release via MAPK. This significantly improves tissue regeneration in diabetic skin infections, highlighting the promising potential of this multifunctional system.
88Development and evaluation of gelatin/hyaluronic acid nanofibrous dressing loaded with silver nanoparticles and phenytoin for enhanced wound healing: an and study.PubMed
Jaber Emami, Niloofar Mostolizadeh, Majid Tabbakhian, et al.
Res Pharm Sci. 2025 Aug 25;20(4):610-634. doi: 10.4103/RPS.RPS_60_25. eCollection 2025 Aug.
BACKGROUND AND PURPOSE: Wound dressings are essential in managing chronic wounds like pressure ulcers, which increase healthcare costs and hospital stays. There is a rising demand for advanced dressings that effectively promote healing. This study developed electrospun gelatin-hyaluronic acid (Gel/HA) nanofibers loaded with silver nanoparticles (Ag NPs) and phenytoin to enhance wound healing. EXPERIMENTAL APPROACH: Ag NPs were synthesized via silver nitrate reduction using trisodium citrate and tannic acid, and characterized for size, zeta potential, PDI, UV-Vis absorption, and XRD patterns. Drug-free and drug-loaded Gel/HA nanofibers were fabricated and analyzed using FE-SEM, FTIR, DSC, XRD, swelling behavior, drug loading, and release profiles. In vitro antibacterial and in vivo wound healing studies were conducted. FINDINGS/RESULTS: Optimized Ag NPs had a size of 41.96 ± 1.2 nm, zeta potential of -23.77 ± 1.31 mV, and PDI of 0.35 ± 0.02. The ideal nanofiber formulation (20 g Gel and 0.25 g HA/100 mL) showed drug loading efficiencies of 56.02 ± 1.8% (Ag NPs) and 61.02 ± 2.82% (phenytoin), with release times of 22.23 and 28.53 h, respectively. The nanofibers demonstrated high swelling (822.2%) and strong antibacterial activity. In vivo studies revealed significantly faster wound closure, improved epithelialization, collagen deposition, and complete healing within 15 days. These effects reflect the synergy between Ag NPs' antimicrobial and phenytoin's regenerative properties. CONCLUSION AND IMPLICATIONS: Gel/HA nanofibers loaded with Ag NPs and phenytoin show great promise as advanced wound dressings. Further studies in larger animal models and clinical trials are warranted.
89Injectable and Self-Healing Probiotics-Loaded Hydrogel for Promoting Superbacteria-Infected Wound Healing.PubMed
Li Mei, Dongjie Zhang, Huarong Shao, et al.
ACS Appl Mater Interfaces. 2022 May 11;14(18):20538-20550. doi: 10.1021/acsami.1c23713. Epub 2022 Apr 26.
Superbacteria-induced skin wound infections are huge health challenges, resulting in significant financial and medical costs due to notable morbidity and mortality worldwide. Probiotics are found in the skin and are effective in treating bacterial infection, moderating the microbial dysbiosis and inflammation induced by pathogens, regulating the immune system, as well as even promoting tissue repair. However, improving their colonization efficiency and viability remains a large obstacle for proper applications. Inspired by probiotic therapy and the natural extracellular matrix structure, hyaluronate-adipic dihydrazide/aldehyde-terminated Pluronic F127/fucoidan hydrogels loaded with (HPF@L.rha) with unique (bio)physicochemical characteristics were developed through the dynamic Schiff-base reaction for superbacteria-infected trauma management. The developed HPF@L.rha exhibit a shortened gelation time, enhanced mechanical strength, and excellent self-healing and liquid-absorption abilities. Importantly, their anti-superbacteria () effect was greatly increased in a dose-dependent fashion. Additionally, evaluation shows that the prepared HPF@L.rha containing appropriate probiotic concentrations (less than 1 × 10 CFU/mL) possess satisfactory cytocompatibility and blood compatibility. Further, compared to the HPF hydrogel, the hydrogel combined with probiotics significantly inhibits infection and inflammation, promotes the formation of re-epithelialization and collagen, and thus accelerates full-thickness superbacteria-infected wound repair, which is comparable to commercial Prontosan gel formulation. This work suggests that the combination of biomimicking hydrogels and probiotic therapy displays the great potential to manage superbug-infected trauma.
90Novel Biocompatible Hyaluronic Acid (HA)/Polyvinylpyrrolidone (PVP) Composites Containing Silver Decorated-Zinc MOF Nanoparticles: Antimicrobial Activity, Drug Delivery and Wound Healing.PubMed
Eman Abdelhakeem, Sally A Mohamed, Hanan H Beherei, et al.
AAPS PharmSciTech. 2025 Oct 30;27(1):5. doi: 10.1208/s12249-025-03243-z.
Chronic infected wounds with multi-drug resistant pathogens present severe therapeutic challenges, often leading to prolonged morbidity and impaired healing. To address this, we engineered a multifunctional wound dressing by integrating Voriconazole-loaded silver/zinc MOFs (VOR@Ag/Zn-MOFs) into a hyaluronic acid/polyvinylpyrrolidone (HA/PVP) matrix. The composite was prepared through sequential steps, including preparing a HA/PVP polymer blend via solvent dissolution, synthesizing bimetallic Ag/Zn-MOFs through chemical precipitation, loading VOR into MOF nanocages, and consolidating the composite via chemical crosslinking followed by freeze-drying. Comprehensive physicochemical characterization to confirm MOF stability was implemented through SEM/EDX, confirming uniform nanocage architecture and elemental distribution (Ag/Zn/F), while DLS revealed controlled particle dimensions, and optimal colloidal stability (PDI 0.694; ZP + 39.3 mV). FTIR confirmed structural integrity through preserved polymer peaks (O-H 3400-3200, C = O 1650 cm⁻1), VOR-specific C-F (1400-1200 cm⁻1), and MOF Zn-O bonds (1100-1000/400-600 cm⁻1), verifying non-covalent integration without degradation. XRD further verified the crystalline framework integrity. In-vitro release studies demonstrated sustained biphasic release (44.2% ± 5.9 burst within 6 h; up to 77.1% ± 1.8 controlled release over 360 h). The composite demonstrated exceptional antimicrobial synergy, including potent antifungal activity against Candida albicans (MFC 1.4 µg/mL, 214-fold lower than free VOR) with microscopically-confirmed cell wall disruption, and broad bacteriostatic activity (MIC range 8,500-17,000 µg/mL against Gram-positive and Gram-negative pathogens). Critically, it achieved complete fungal clearance within 24 h. Simultaneously, it accelerated tissue regeneration, showing outstanding fibroblast biocompatibility (IC₃₀ > 3,000 µg/mL; 6,000-fold safety margin) and enabling marked wound closure within 72 h in the wound healing study. By converging triple-action mechanisms, including metal ion synergy (Ag⁺/Zn2⁺), targeted azole delivery, and HA/PVP-mediated pro-regenerative signaling, this platform uniquely eliminates resilient infections and restores wound integrity that was previously unattainable with conventional azole therapies.
91A synergistically antimicrobial and antioxidant hyaluronic acid hydrogel for infected wounds.PubMed
Yongping Lu, Weiqi Kang, Yue Yu, et al.
Int J Biol Macromol. 2024 Jun;269(Pt 2):131795. doi: 10.1016/j.ijbiomac.2024.131795. Epub 2024 Apr 24.
Bacterial infections during wound healing impede the healing process and trigger local or systemic inflammatory reactions. Consequently, there is an urgent need to develop a new material with antimicrobial and antioxidant properties to promote infected wound healing. A synergistically antimicrobial and antioxidant hyaluronic acid hydrogel (HMn) is prepared by employing MnO nanosheets into 4ARM-PEG5000-SH crosslinked methacrylated hyaluronic acid (HAMA) network. The coordination between sulfhydryl groups of 4ARM-PEG5000-SH and MnO nanosheets ensures entrapment of the nanosheets within the hydrogel, while the interaction between 4ARM-PEG5000-SH and HAMA results in facile gelation through thiol-ene click reaction. MnO nanosheets exhibit strong photothermal properties and reactive oxygen species (ROS) scavenging abilities, while hyaluronic acid promotes wound healing. When subjected to near-infrared (NIR) irradiation, the HMn achieves a bactericidal rate of 95.24 % for Staphylococcus aureus and nearly 100 % for Escherichia coli. In animal experiments, treatment with the HMn under NIR irradiation results in the best wound healing outcomes. Both in vitro and vivo biocompatible assays demonstrate that the HMn has rarely cell cytotoxicity and tissue damage. The HMn is easy to prepare and has good biocompatibility as well as efficient antibacterial and antioxidant properties, providing a novel method for the treatment of infected wounds.
92Living Bacterial Hydrogels for Accelerated Infected Wound Healing.PubMed
Zunzhen Ming, Lin Han, Meiyu Bao, et al.
Adv Sci (Weinh). 2021 Dec;8(24):e2102545. doi: 10.1002/advs.202102545. Epub 2021 Oct 31.
Damaged skin cannot prevent harmful bacteria from invading tissues, causing infected wounds and even serious tissue damage. Traditional treatments can not only kill pathogenic bacteria, but also suppress the growth of beneficial bacteria, thus destroying the balance of the damaged skin microbial ecosystem. Here, a living bacterial hydrogel scaffold is reported that accelerates infected wound healing through beneficial bacteria secreting antibacterial substances. Lactobacillus reuteri, a common probiotic, is encapsulated in hydrogel microspheres by emulsion polymerization and further immobilized in a hydrogel network by covalent cross-linking of methacrylate-modified hyaluronic acid. Owing to light-initiated crosslinking, the hydrogel dressing can be generated in situ at the wound site. This hydrogel scaffold not only protects bacteria from immune system attack, but also prevents bacteria from escaping into the local environment, thus avoiding potential threats. Both in vitro and in vivo experiments show that it has excellent ability against harmful bacteria and anti-inflammatory capabilities, promoting infected wound closure and new tissue regeneration. This work may open up new avenues for the application of living bacteria in the clinical management of infected wounds.
93Multifunctional Therapeutic Nanodiamond Hydrogels for Infected-Wound Healing and Cancer Therapy.PubMed
Peiwen Wang, Zishuo Hou, Zizhen Wang, et al.
ACS Appl Mater Interfaces. 2024 Feb 28;16(8):9656-9668. doi: 10.1021/acsami.3c13464. Epub 2024 Feb 20.
Wound infection and tumor recurrence are the two main threats to cancer patients after surgery. Although researchers have developed new treatment systems to address the two significant challenges simultaneously, the potential side effects of the heavy-metal-ion-based treatment systems still severely limit their widespread application in therapy. In addition, the wounds from tumor removal compared with general operative wounds are more complex. The tumor wounds mainly exhibit more hemorrhage, larger trauma area, greater vulnerability to bacterial infection, and residual tumor cells. Therefore, a multifunctional treatment platform is urgently needed to integrate rapid hemostasis, sterilization, wound healing promotion, and antitumor functions. In this work, nanodiamonds (NDs), a material that has been well proven to have excellent biocompatibility, are added into a solution of acrylic-grafted chitosan (CEC) and oxidized hyaluronic acid (OHA) to construct a multifunctional treatment platform (CEC-OHA-NDs). The hydrogels exhibit rapid hemostasis, a wound-healing-promoting effect, excellent self-healing, and injectable abilities. Moreover, CEC-OHA-NDs can effectively eliminate bacteria and inhibit tumor proliferation by the warm photothermal effect of NDs under tissue-penetrable near-infrared laser irradiation (NIR) without cytotoxicity. Consequently, we adopt a simple and convenient strategy to construct a multifunctional treatment platform using carbon-based nanomaterials with excellent biocompatibility to promote the healing of infected wounds and to inhibit tumor cell proliferation simultaneously.
94Macrophage polarization and diabetic wound healing.PubMed
Amanda E Louiselle, Stephen M Niemiec, Carlos Zgheib, et al.
Transl Res. 2021 Oct;236:109-116. doi: 10.1016/j.trsl.2021.05.006. Epub 2021 Jun 2.
Diabetes mellitus is a costly disease and nearly one-third of these costs are attributed to management of diabetic foot disease including chronic, non-healing, diabetic foot ulcers. Therefore, much effort has been placed into understanding the pathogenesis of diabetic wounds and novel therapeutics. A relatively new area of interest has been macrophage polarization and its role in diabetic wound healing. Diabetic wounds show dysregulated and persistent M1 (pro-inflammatory) macrophage polarization whereas normal wounds will display a transition to M2 (pro-healing) macrophages around day three after wounding. We reviewed factors known to affect macrophage polarization, mostly focused on those that contribute to M2 macrophage polarization, and potential treatments that at least in part target macrophage polarization in the diabetic wound bed. Much of the work has been aimed at reducing hyperglycemia and encouraging pro-inflammatory cytokine neutralization or decreased expression given this has a significant role in producing M1 macrophages. Treatment of diabetic wounds will likely require a multi-modal approach including management of underlying diabetes and control of hyperglycemia, topical therapeutics, and prevention of secondary infection and inflammation.
95Single cell transcriptomic landscape of diabetic foot ulcers.PubMed
Georgios Theocharidis, Beena E Thomas, Debasree Sarkar, et al.
Nat Commun. 2022 Jan 10;13(1):181. doi: 10.1038/s41467-021-27801-8.
Diabetic foot ulceration (DFU) is a devastating complication of diabetes whose pathogenesis remains incompletely understood. Here, we profile 174,962 single cells from the foot, forearm, and peripheral blood mononuclear cells using single-cell RNA sequencing. Our analysis shows enrichment of a unique population of fibroblasts overexpressing MMP1, MMP3, MMP11, HIF1A, CHI3L1, and TNFAIP6 and increased M1 macrophage polarization in the DFU patients with healing wounds. Further, analysis of spatially separated samples from the same patient and spatial transcriptomics reveal preferential localization of these healing associated fibroblasts toward the wound bed as compared to the wound edge or unwounded skin. Spatial transcriptomics also validates our findings of higher abundance of M1 macrophages in healers and M2 macrophages in non-healers. Our analysis provides deep insights into the wound healing microenvironment, identifying cell types that could be critical in promoting DFU healing, and may inform novel therapeutic approaches for DFU treatment.
96Snail-inspired AFG/GelMA hydrogel accelerates diabetic wound healing via inflammatory cytokines suppression and macrophage polarization.PubMed
Zhipeng Zhou, Tuo Deng, Maixian Tao, et al.
Biomaterials. 2023 Aug;299:122141. doi: 10.1016/j.biomaterials.2023.122141. Epub 2023 May 7.
Diabetic foot ulcers (DFUs) are a severe and rapidly growing diabetic complication, but treating DFUs remains a challenge for the existing therapies are expensive and highly non-responsive. Recently, we discovered that a natural adhesive from snail mucus can promote skin wound healing. Herein, inspired by the finding, we developed a double-network hydrogel biomaterial that composed of snail glycosaminoglycan (AFG) and methacrylated gelatin (GelMA), in which AFG is the main bioactive component of snail mucus and GelMA provides a scaffold mimicking the proteins in snail mucus. The biomimetic hydrogel exhibited strong tissue adhesion, potent anti-inflammatory activity, and excellent biocompatibility. The biodegradable AFG/GelMA hydrogel markedly promoted chronic wound healing in both STZ-induced type 1 diabetic rat and db/db mouse models after a single treatment. Further mechanistic research showed that the hydrogel significantly attenuated inflammation by sequestrating pro-inflammatory cytokines, as well as downregulated their expression by inhibiting NF-ĸB signaling pathway, and it can also promote macrophage polarization to M2 phenotype. Taken together, the bioinspired hydrogel can effectively promote the transition of chronic wounds from inflammation to proliferation stage. These data suggest that the AFG/GelMA hydrogel is a promising therapeutic biomaterial for the treatment of chronic diabetic wounds.
97An antibacterial, antioxidant and hemostatic hydrogel accelerates infectious wound healing.PubMed
Ziyi Zhou, Dengjun Zhang, Xuchao Ning, et al.
J Nanobiotechnology. 2025 Jan 28;23(1):49. doi: 10.1186/s12951-025-03148-w.
Hydrogel drug-delivery system that can effectively load antibacterial drugs, realize the in-situ drug release in the microenvironment of wound infection to promote wound healing. In this study, a multifunctional hydrogel drug delivery system (HA@TA-Okra) was constructed through the integration of hyaluronic acid methacrylate (HAMA) matrix with tannic acid (TA) and okra extract. The composition and structural characteristics of HA@TA-Okra system and its unique advantages in the treatment of diverse wounds were systematically evaluated. TA, due to its unique chemical structure, is able to anchor within the HAMA network through interactions and cross-linking, conferring exceptional mechanical strength and stability to the hydrogel. Both TA and okra extract possess antioxidant and antibacterial properties, and when they two acts synergistically they can effectively scavenge free radicals, enhance antibacterial action, diminishing the risk of wound infection. In vitro experiments revealed that HA@TA-Okra system has superior properties, such as rapid gel response, remarkable swelling regulation, and potent antioxidant ability. Furthermore, the HA@TA-Okra system significantly outperformed conventional dressings in terms of hemostatic performance in a rat hemorrhage model. We further evaluated the repair role of HA@TA-Okra system in vivo by establishing an animal model of full-thickness skin defects and a model of infected total skin defects. The results confirmed its positive effects in fighting bacterial infection, reducing inflammation and promoting wound healing. In summary, the HA@TA-Okra system exhibits comprehensive properties such as antibacterial, antioxidant and hemostatic properties, which has a potential application in the field of tissue repair medicine.
98Different molecular weights of hyaluronan research in knee osteoarthritis: A state-of-the-art review.PubMed
Ko-Hsiu Lu, Peace Wun-Ang Lu, Chiao-Wen Lin, et al.
Matrix Biol. 2023 Mar;117:46-71. doi: 10.1016/j.matbio.2023.02.006. Epub 2023 Feb 26.
Osteoarthritis (OA), the most common form of arthritis, is characterized by progressive cartilage destruction, concomitant adaptive osteogenesis, and loss of joint function. The progression of OA with aging is associated with a decrease in native hyaluronan (HA, hyaluronate or hyaluronic acid) with a high molecular weight (HMW) in synovial fluid and a subsequent increase in lower MW HA and fragments. As HMW HA possesses numerous biochemical and biological properties, we review new molecular insights into the potential of HA to modify OA processes. Different MWs in the formulation of products appear to have varying effects on knee OA (KOA) pain relief, improved function, and postponing surgery. In addition to the safety profile, more evidence indicates that intraarticular (IA) HA administration may be an effective option to treat KOA, with a particular emphasis on the use of HA with fewer injections of higher MW, including potential applications of HA of very HMW. We also analyzed published systemic reviews and meta-analyses of IA HA in treating KOA in order to discuss their conclusions and consensus statements. According to its MW, HA may offer a simple way to refine therapeutic information in selective KOA.
99Cartilage boundary lubrication of ovine synovial fluid following anterior cruciate ligament transection: a longitudinal study.PubMed
M Atarod, T E Ludwig, C B Frank, et al.
Osteoarthritis Cartilage. 2015 Apr;23(4):640-7. doi: 10.1016/j.joca.2014.12.017. Epub 2014 Dec 30.
OBJECTIVE: To assess ovine synovial fluid (oSF) from different post-injury time points for (1) proteoglycan-4 (PRG4) and hyaluronan (HA) concentration, (2) HA molecular weight (MW) distribution, (3) cartilage boundary lubrication function, and (4) lubricant composition-function relationships. The association between cartilage boundary lubrication and gross cartilage changes after injury was also examined. METHODS: oSF was collected 2, 4, 10, and 20 weeks post anterior cruciate ligament (ACL) transection in five skeletally mature sheep. PRG4 and HA concentrations were measured using sandwich enzyme-linked immunosorbent assay, and HA MW distribution by agarose gel electrophoresis. Cartilage boundary lubrication of oSF was assessed using a cartilage-cartilage friction test. Gross damage to articular cartilage was also quantified at 20 weeks using modified Drez scoring protocol. RESULTS: Early (2-4 weeks) after ACL injury, PRG4 concentrations were significantly higher (P = 0.045, P = 0.037), and HA concentrations were substantially lower (P = 0.005, P = 0.005) compared to 20 weeks. The HA MW distribution also shifted towards lower ranges in the early post-injury stage. The kinetic friction coefficients were significantly higher 2-4 weeks post injury (P = 0.008 and P = 0.049) compared to 20 weeks. Poor cartilage boundary lubricating ability early after injury was associated with cartilage damage at 20 weeks. CONCLUSION: Altered composition and diminished boundary lubrication of oSF early after ACL transection may pre-dispose the articular cartilage to degenerative changes and initiate osteoarthritis (OA). These observations also provide potential motivation for biotherapeutic interventions at earlier time points post injury.
100High-Molecular-Weight Hyaluronic Acid Inhibits IL-1β-Induced Synovial Inflammation and Macrophage Polarization through the GRP78-NF-κB Signaling Pathway.PubMed
Chien-Hsing Lee, Chi-Fu Chiang, Feng-Chih Kuo, et al.
Int J Mol Sci. 2021 Nov 3;22(21):11917. doi: 10.3390/ijms222111917.
Recent evidence has suggested that synovial inflammation and macrophage polarization were involved in the pathogenesis of osteoarthritis (OA). Additionally, high-molecular-weight hyaluronic acid (HMW-HA) was often used clinically to treat OA. GRP78, an endoplasmic reticulum (ER) stress chaperone, was suggested to contribute to the hyperplasia of synovial cells in OA. However, it was still unclear whether HMW-HA affected macrophage polarization through GRP78. Therefore, we aimed to identify the effect of HMW-HA in primary synovial cells and macrophage polarization and to investigate the role of GRP78 signaling. We used IL-1β to treat primary synoviocytes to mimic OA, and then treated them with HMW-HA. We also collected conditioned medium (CM) to culture THP-1 macrophages and examine the changes in the phenotype. IL-1β increased the expression of GRP78, NF-κB (p65 phosphorylation), IL-6, and PGE2 in primary synoviocytes, accompanied by an increased macrophage M1/M2 polarization. GRP78 knockdown significantly reversed the expression of IL-1β-induced GRP78-related downstream molecules and macrophage polarization. HMW-HA with GRP78 knockdown had additive effects in an IL-1β culture. Finally, the synovial fluid from OA patients revealed significantly decreased IL-6 and PGE2 levels after the HMW-HA treatment. Our study elucidated a new form of signal transduction for HMW-HA-mediated protection against synovial inflammation and macrophage polarization and highlighted the involvement of the GRP78-NF-κB signaling pathway.
101The relationship of ALPK1, hyaluronic acid and M1 macrophage polarization in the temporomandibular joint synovitis.PubMed
Jie Zhao, Yaping Feng, Xin Liu, et al.
J Cell Mol Med. 2024 Apr;28(7):e18172. doi: 10.1111/jcmm.18172.
M1 macrophage polarization and synovitis play an important role in the pathogenesis of temporomandibular joint osteoarthritis (TMJOA). Reduced molecular weight of hyaluronic acid (HA) in synovial fluid of patients with TMJOA. In addition, high molecular weight hyaluronic acid (HMW-HA) is often used clinically to treat TMJ inflammation. As a pattern recognition receptor of the cytoplasm, ALPK1 was found to be pro-inflammatory in a variety of diseases. However, the relationship of ALPK1, HA and M1 macrophage polarization in TMJ synovitis remains unclear. We aimed to investigate the role of ALPK1 and HA in macrophage polarization and TMJ synovitis and the underlying mechanisms. The results demonstrated that ALPK1 was highly upregulated in the synovial macrophages in the inflamed TMJ synovium of patients. Low molecular weight hyaluronic acid (LMW-HA) promoted the expression of ALPK1 and M1 macrophage-associated genes. Besides, rhALPK1 promoted the expression of M1 macrophage-associated factors and the nuclear translocation of PKM2. Furthermore, ALPK1 knockout mice exhibited limited infiltration of macrophages and decreased expression levels of M1 macrophage-associated genes in CFA-induced TMJ synovitis. While HMW-HA inhibited the expression of ALPK1 and M1 macrophage polarization. Our results elucidated that ALPK1 promoted TMJ synovitis by promoting nuclear PKM2-mediated M1 macrophage polarization, whereas HMW-HA inhibited the expression of ALPK1 as well as M1 macrophage polarization.
102Hyaluronic Acid Hampers the Inflammatory Response Elicited by Extracellular Vesicles from Activated Monocytes in Human Chondrocytes.PubMed
Vittoria Carrabs, Maria Isabel Guillén, María Luisa Ferrándiz, et al.
Pharmaceutics. 2024 Oct 28;16(11):1386. doi: 10.3390/pharmaceutics16111386.
Osteoarthritis (OA) is the most common joint disease in the adult population. OA is the result of multiple mechanisms leading to inflammation and the degradation of the cartilage. A complex series of etiological actors have been identified so far, including extracellular vesicles (EVs). The EV content of the synovial fluid (SF) can release inflammatory mediators that enhance OA progression. An intra-articular viscosupplementation of high-MW hyaluronic acid (HyA) constitutes the first-line conservative treatment for OA. Although attractive for the potential pharmacological implications, the possibility that HyA may interact with EVs in the context of OA has not yet been specifically investigated; therefore, the present study aimed to fill this gap. We studied the effect of a HyA preparation (a blend of crosslinked and linear polymers, CLHyA) on the relevant inflammatory markers in chondrocytes (HC cells or primary chondrocytes isolated from patients with advanced OA) exposed to the EVs collected from IL-1β-stimulated THP-1 human monocytes (EVs+). EVs+ caused specific inflammatory responses in chondrocytes that could be prevented by coincubation with CLHyA. This anti-inflammatory activity is likely dependent on the direct binding of CLHyA to CD44 receptors highly expressed in EVs+ and on the subsequent hindrance to EVs+ diffusion and docking to target cells. On the whole, the tight interactions identified herein between HMW HyA and EVs+ represent a novel, pharmacologically exploitable mechanism potentially relevant in the context of OA treatment.
103Safety and efficacy of a single intra-articular injection of hyaluronic acid in osteoarthritis of the hip: a case series of 87 patients.PubMed
David M Long, Jane Fitzpatrick
BMC Musculoskelet Disord. 2021 Sep 16;22(1):797. doi: 10.1186/s12891-021-04672-0.
BACKGROUND: Osteoarthritis (OA) is the most prevalent form of joint disease and commonly affects the hip. Hip OA is associated with a high socioeconomic burden. Intra-articular hyaluronic acid (HA) injection may be of benefit but quality evidence for HA use in hip OA is lacking. The purpose of this study was to assess the safety and efficacy of ultrasound guided injection of a high molecular weight, non-animal derived, stabilised HA (NASHA) in patients with mild to moderate hip OA. METHODS: This single site study is an analysis of prospectively collected outcome data for 87 consecutive patients over a 2-year period who received a single HA (Durolane) injection for symptomatic hip OA. Inclusion criteria were male or female patients over 18-years of age with mild to moderate hip OA on x-ray. Patients with severe hip OA were excluded. The primary outcome measure was a modified Harris Hip Score (mHHS) questionnaire at baseline and 6-weeks with a minimal clinically important difference (MCID) of 10 points. All adverse events were recorded and assessed. RESULTS: Data from 87 patients, 49 women and 38 men with mean age of 54 (SD = 10.8) were analysed. At baseline, mean mHHS was 58.47 (SD 14.31). At the 6 week follow up, mean mHHS improved to 71.30 (SD 16.46), a difference of 12.83 (p < 0.01). This was greater than the MCID of 10. No significant adverse events were encountered. Five patients reported short-lived injection site pain. CONCLUSION: A single injection of HA (NASHA) in the setting of hip joint OA was both safe and efficacious in this 87 patient cohort. Improvement in pain and function as measured with mHHS was statistically significant and reached the MCID of 10. TRIAL REGISTRATION: The study was retrospectively registered on the 1st of February 2021 in the Australian New Zealand Clinical Trials Registry with registry number ACTRN12621000098831 . All research was performed in accordance with the Declaration of Helsinki.
104A Multicenter, Randomized, Double-Blinded, Placebo-Controlled Clinical Trial to Evaluate the Efficacy and Safety of a Krill Oil, Astaxanthin, and Oral Hyaluronic Acid Complex on Joint Health in People with Mild Osteoarthritis.PubMed
W Stephen Hill, Margaret H Dohnalek, Yejin Ha, et al.
Nutrients. 2023 Aug 29;15(17):3769. doi: 10.3390/nu15173769.
Osteoarthritis is a significant global health problem. Many patients seek more effective alternatives to nonsteroidal anti-inflammatory medicines or commercial supplements to manage joint pain and inflammation. FlexPro MD (FP-MD) combines krill oil, astaxanthin, and lower molecular weight hyaluronic acid to support joint health. A 12-week, randomized, double-blind, placebo-controlled trial compared the efficacy and safety of FP-MD and placebo once daily in participants (n = 100) with mild osteoarthritis of the knee or hip joint. For the primary endpoint of joint pain score, per-protocol participants (n = 75) in the FP-MD group (n = 37) had a statistically significantly greater mean reduction from baseline in the Korean Visual Analog Scale (K-VAS) at week 12 compared with participants in the placebo group (n = 38) (20.8 ± 16.16 mm vs. 10.6 ± 17.58, = 0.0105). The Korean Western Ontario and McMaster Universities Osteoarthritis Index (K-WOMAC) total score was also significantly improved in the FP-MD group at week 12 compared with placebo (-13.0 ± 13.62 vs. -5.5 ± 18.08, = 0.0489), especially an improvement in pain score (-2.5 ± 2.92 vs. -1.3 ± 3.94, = 0.02635). FP-MD group had greater improvement in joint function scoring by investigator assessment ( = 0.0127) and by group participants ( = 0.0070). A statistically significantly greater number of patients reported adverse events in the placebo group compared with the FP-MD group (16% vs. 4%, = 0.0455), most commonly gastrointestinal disorders in both of the groups. These findings suggest that FP-MD is well tolerated and can be effectively used to address joint pain in patients diagnosed with mild osteoarthritis, the main symptom of this condition.
105Addition of High Molecular Weight Hyaluronic Acid to Fibroblast-Like Stromal Cells Modulates Endogenous Hyaluronic Acid Metabolism and Enhances Proteolytic Processing and Secretion of Versican.PubMed
Jiapeng Xue, Jinnan Chen, Quan Shen, et al.
Cells. 2020 Jul 13;9(7):1681. doi: 10.3390/cells9071681.
We have examined the effect of exogenous linear chain high molecular weight hyaluronic acid (HMW HA) on endogenously synthesized hyaluronic acid (HA) and associated binding proteins in primary cultures of fibroblast-like stromal cells that were obtained by collagenase digestion of the murine peripatellar fat pad. The cultures were expanded in DMEM that was supplemented with fetal bovine serum and basic fibroblast growth factor (bFGF) then exposed to macrophage-colony-stimulating factor (MCSF) to induce macrophage properties, before activation of inflammatory pathways using lipopolysaccharide (LPS). Under all culture conditions, a significant amount of endogenously synthesized HA localized in LAMP1-positive lysosomal vesicles. However, this intracellular pool was depleted after the addition of exogenous HMW HA and was accompanied by enhanced proteolytic processing and secretion of de novo synthesized versican, much of which was associated with endosomal compartments. No changes were detected in synthesis, secretion, or proteolytic processing of aggrecan or lubricin (PRG4). The addition of HMW HA also modulated a range of LPS-affected genes in the TLR signaling and phagocytosis pathways, as well as endogenous HA metabolism genes, such as , , and . However, there was no evidence for association of endogenous or exogenous HMW HA with cell surface CD44, TLR2 or TLR4 protein, suggesting that its physiochemical effects on pericelluar pH and/or ionic strength might be the primary modulators of signal transduction and vesicular trafficking by this cell type. We discuss the implications of these findings in terms of a potential in vivo effect of therapeutically applied HMW HA on the modification of osteoarthritis-related joint pathologies, such as pro-inflammatory and degradative responses of multipotent mesenchymal cells residing in the synovial membrane, the underlying adipose tissue, and the articular cartilage surface.
106The role of high molecular weight hyaluronic acid in mucoadhesion on an ocular surface model.PubMed
Cristian Guarise, Laura Acquasaliente, Gianfranco Pasut, et al.
J Mech Behav Biomed Mater. 2023 Jul;143:105908. doi: 10.1016/j.jmbbm.2023.105908. Epub 2023 May 15.
Hyaluronic acid (HA) is frequently formulated in eye drops to improve the stability of the tear film by hydration and lubrication. Mucoadhesion is related to the ocular residence time and therefore to the effectiveness of the eye drops. The ocular residence time of the HA formulation is correlated with the ability of HA to create specific strong interactions in the ocular surface with the mucus layer, mainly composed of a mixture of secreted mucins (MUC; gel forming MUC5AC and MUC2) and shed membrane-bound soluble mucins (MUC1, MUC4, and MUC16). Dry eye disease (DED) is a multifactorial pathology of the preocular tear film with possible damage to the ocular surface classified in two types: (1) aqueous-deficient dry eye and (2) evaporative dry eye, caused by a decrease in goblet cell density that reduces MUC expression and/or by meibomian gland dysfunction, that results in a drop in the lipidic fraction of the tear film. In this work, the binding affinity between HA and MUC2 has been evaluated with three complementary approaches because the secreted MUCs play a pivotal role in the viscoelastic properties of the tear film: 1. Rheological analysis, measuring the mucoadhesive index and the complex viscosity in relation to MM (Molecular Mass) and concentration; 2. Fluorescence analysis, using a fluorescent hydrophobic probe, to investigate the conformational change of MUC2 during the interaction with the HA polymer; 3. Surface plasmon resonance analysis, used to measure the affinity between MUC2 (immobilized on the surface of a sensor chip) and the HA polymers that flowed on it at the molecular level. For all these tests, the mucoadhesive performance of the natural HA linearly increases with the MM, whereas cross-linked HA and other emollient and gelling agents (formulated in artificial tears) do not show the same mucoadhesive properties (with the exception of xanthan gum). The mucoadhesive performance of high MM HA has also been confirmed in conditions that simulate the pathological condition of the tear film during DED by decreasing the MUC2 or oleic acid concentration. Physico-chemical analysis of a series of marketed artificial tears confirms the linear correlation between the MM of the HA used in the products and the mucoadhesive index measured on the ocular surface model.
107The Effects of High Molecular Weight Hyaluronic Acid Eye Drop Application in Environmental Dry Eye Stress Model Mice.PubMed
Takashi Kojima, Taeko Nagata, Haruka Kudo, et al.
Int J Mol Sci. 2020 May 15;21(10):3516. doi: 10.3390/ijms21103516.
Hyaluronic acid (HA) ophthalmic solution is widely used in dry eye treatment worldwide. However, there are no reports comparing the dry eye treatment effects of high molecular weight HA with low molecular weight HA. Sixty eight-week-old C57BL/6 mice were assigned to the following 6 groups and exposed to environmental dry eye stress (EDES) that mimics office work environment: 1) 0.1% low molecular weight HA (LMWHA) eye drops, 2) 0.3% LMWHA eye drops, 3) 3% diquafosol sodium (DQ) eye drops, 4) 0.15% high molecular weight HA (HMWHA) eye drops, 5) no treatment with exposure to EDES (EDES+/Treatment-), and 6) no treatment without exposure to EDES (EDES-/Treatment-). After EDES, the HMWHA group had significantly longer break-up time (BUT) than the 0.1%, 0.3% LMWHA groups and the DQ group. After EDES, the HMWHA group had significantly lower lissamine green staining scores than the LMWHA and DQ groups. Subepithelial presumed dendritic cell density in the HMWHA group was significantly lower than the EDES+/Treatment- group. After EDES exposure, Conjunctival Muc5AC mRNA expression in the HMWHA group was significantly higher than the 0.1 and 0.3% LMWHA groups. Ophthalmic HMWHA solution may have a better dry eye treatment effect than LMWHA or DQ solution, owing to its anti-inflammatory effect.
108Management of Oxidative Stress and Inflammation in Patients with Symptomatic Dry Eye Disease Treated with a Preservative-Free Ophthalmic Emulsion Combining Alpha-Lipoic Acid and High Molecular Weight Sodium Hyaluronate.PubMed
Laure Chauchat, Camille Guerin, Marwan Sahyoun, et al.
Adv Ther. 2025 May;42(5):2219-2233. doi: 10.1007/s12325-025-03139-1. Epub 2025 Mar 13.
INTRODUCTION: This study aimed to explore the beneficial effect of a preservative-free (PF) emulsion eyedrop combining high molecular weight sodium hyaluronate (HMW-HA) and alpha-lipoic acid on oxidative stress and ocular surface inflammation in patients with dry eye disease (DED). METHODS: In this prospective exploratory study, patients with moderate to severe DED symptoms were treated with the study eyedrop, 4-6 times/day for 1 month. Two visits were scheduled: baseline (D0) and after 35 days (D35). The following parameters were assessed: quality of life (OSDI score); superoxide dismutase (SODase) concentration in tears; goblet cell (GC) density (impression cytology); conjunctival hyperemia (Efron scale); corneal, conjunctival, and eyelid staining (Oxford scale); and meibomian gland (MG) blockage. Data were compared between D0 and D35 in subgroups of patients with a significant abnormality of the study outcome at baseline. RESULTS: Forty patients were involved, with a highly significant improvement of OSDI score at D35. The mean concentration of SODase significantly increased by 3.2 and 2.4 times for SODase1 and SODase2 respectively, in patients with deficient SODase at baseline. In patients with abnormal GC density at baseline, GC count increased by five times at D35. Conjunctival hyperemia and corneal staining scores significantly improved in the subpopulation of patients with a baseline grade ≥ 2. Eyelid margin staining was significantly reduced at D35 in patients with significant abnormalities at baseline. A significant reduction of MG obstruction was shown in the lower eyelid in patients with significant blockage at baseline. CONCLUSION: The use of a PF ophthalmic emulsion combining alpha-lipoic acid and HMW-HA shows a beneficial effect on the ocular surface through the improved quality of life score. Noteworthy, patients presenting high oxidative and inflammatory conditions experienced significant improvement in oxidative stress and inflammation markers; this synergic effect is likely due to the well-known properties of alpha-lipoic acid and HMW-HA, enhancing DED management. TRIAL REGISTRATION: ISRCTN.com identifier, 17861788.
109Sodium Hyaluronate-PDGF Repairs Cartilage and Subchondral Bone Microenvironment via HIF-1α-VEGF-Notch and SDF-1-CXCR4 Inhibition in Osteoarthritis.PubMed
Zhengchao Wang, Pengfei Zhu, Hongmei Li, et al.
J Cell Mol Med. 2025 Apr;29(7):e70515. doi: 10.1111/jcmm.70515.
Chronic degenerative changes in cartilage and subchondral bone that lead to instability of the cartilage microenvironment are essential for the development of osteoarthritis (OA) in the old. Synchronous repair of cartilage and subchondral bone may be a key strategy for OA treatment. PDGF-BB effectively promoted chondrocyte regeneration and angiogenesis. However, the mechanisms by which PDGF-BB affects subchondral bone and the delivery of PDGF-BB to the joint cavity need to be further explored. In this study, we used sodium hyaluronate to deliver PDGF-BB (SH-PDGF) to the joint space and aimed to determine the mechanisms of SH-PDGF in repairing cartilage and subchondral bone and stabilising the cartilage microenvironment. In this research, we determined the pharmacokinetics of PDGF-BB and SH-PDGF in cartilage. Moreover, we investigated the effects of PDGF-BB and SH-PDGF on cartilage and the subchondral bone microenvironment by identifying changes in the HIF-VEGF-Notch axis and SDF-1-CXCR4 axis in an OA rat model. The results showed that PDGF-BB increased cell viability, decreased HIF-1α levels, inhibited inflammation and improved matrix metabolism in osteoarthritic chondrocytes under hyperoxic or hypoxic conditions. We also found that PDGF-BB and SH-PDGF showed similar effects on repairing cartilage and subchondral bone simultaneously. However, SH-PDGF had some advantages over PDGF-BB in prolonging the injection interval and decreasing the injection time. These protective effects were mediated by the inhibition of both the HIF-1α-VEGF-Notch axis and the SDF-1-CXCR4 axis. The underlying mechanisms include the inhibition of HIF-1α-VEGF-Notch-mediated vessel invasion and SDF-1-CXCR4 axis-mediated crosstalk between cartilage and subchondral tissue.
110Intra-articular injection of synovium-derived mesenchymal stem cells and hyaluronic acid promote regeneration of massive cartilage defects in rabbits.PubMed
Vyacheslav Ogay, Miras Karzhauov, Ainur Mukhambetova, et al.
Cent Asian J Glob Health. 2014 Jan 24;2(Suppl):97. doi: 10.5195/cajgh.2013.97. eCollection 2013.
INTRODUCTION: The purpose of this study was to investigate whether intra-articular injection of synovium-derived mesenchymal stem cells (SD MSCs) with low molecular weight hyaluronic acid (HA) could promote regeneration of massive cartilage in rabbits. MATERIAL AND METHODS: The SD MSCs were harvested from the knees of 10 Flemish giant rabbits, expanded in culture, and characterized. A reproducible 4-mm cylindrical defect was created in the intercondylar groove area using a kit for the mosaic chondroplasty of femoral condyle COR (De Puy, Mitek). The defect was made within the cartilage layer without destruction of subchondral bone. Two weeks after the cartilage defect, SD MSCs (2 × 106 cell/0.15 ml) were suspended in 0.5% low molecular weight HA (0.15 ml) and injected into the left knee, and HA solution (0.30 ml) alone was placed into the right knee. Cartilage regeneration in the experimental and control groups were evaluated by macroscopically and histologically at 10, 30, and 60 days. RESULTS: On day 10, after intra-articular injection of SD MSCs, we observed an early process of cartilage regeneration in the defect area. Histological studies revealed that cartilage defect was covered by a thin layer of spindle-shaped undifferentiated cells and proliferated chodroblasts. In contrast, an injection of HA did not induce reparation of cartilage in the defect area. At 30 days, macroscopic observation showed that the size of cartilage defect after SD MSC injection was significantly smaller than after HA injection. Histological score was also better in the MSC-treated intercondylar defect. At 60 days after MSC treatment, cartilage defect was nearly nonexistent and looked similar to an intact cartilage. CONCLUSION: Thus, intra-articular injection of SD MSCs can adhere to the defect in the intercondylar area, and promote cartilage regeneration in rabbits.
111A Superparamagnetic Composite Hydrogel Scaffold as In Vivo Dynamic Monitorable Theranostic Platform for Osteoarthritis Regeneration.PubMed
Mayifei Rong, Dingge Liu, Xiaoguang Xu, et al.
Adv Mater. 2024 Aug;36(35):e2405641. doi: 10.1002/adma.202405641. Epub 2024 Jul 6.
Osteoarthritis (OA) is a prevalent disease, characterized by subchondral fractures in its initial stages, which has no precise and specific treatment now. Here, a novel multifunctional scaffold is synthesized by photopolymerizing glycidyl methacrylate-modified hyaluronic acid (GMHA) as the matrix in the presence of hollow porous magnetic microspheres based on hydroxyapatite. In vivo subchondral bone repairing results demonstrate that the scaffold's meticulous design has most suitable properties for subchondral bone repair. The porous structure of inorganic particles within the scaffold facilitates efficient transport of loaded exogenous vascular endothelial growth factor (VEGF). The FeO nanoparticles assembled in microspheres promote the osteogenic differentiation of bone marrow mesenchymal stem cells and accelerate the new bone generation. These features enable the scaffold to exhibit favorable subchondral bone repair properties and attain high cartilage repair scores. The therapy results prove that the subchondral bone support considerably influences the upper cartilage repair process. Furthermore, magnetic resonance imaging monitoring demonstrates that FeO nanoparticles, which are gradually replaced by new bone during osteochondral defect repair, allow a noninvasive and radiation-free assessment to track the newborn bone during the OA repair process. The composite hydrogel scaffold (CHS) provides a versatile platform for biomedical applications in OA treatment.
112Cartilage regeneration using transforming growth factor-beta 3-loaded injectable crosslinked hyaluronic acid hydrogel.PubMed
Ju Hwa Lee, Pil Yun Kim, Yun Chang Pyun, et al.
Biomater Sci. 2024 Jan 16;12(2):479-494. doi: 10.1039/d3bm01008b.
Cartilage defects can be difficult to heal, potentially leading to complications such as osteoarthritis. Recently, a tissue engineering approach that uses scaffolds and growth factors has been proposed to regenerate new cartilage tissues. Herein, we investigated the application of hyaluronic acid (HA) gel loaded with transforming growth factor-beta 3 (TGF-β3) for enhanced cartilage regeneration. We assessed the clinical conditions required to efficiently enhance the ability of the modified HA gel to repair defective cartilage. Based on our findings, the prepared HA gel exhibited good physicochemical and mechanical properties and was non-toxic and non-inflammatory. Moreover, HA gel-loaded TGF-β3 (HAT) had improved biocompatibility and promoted the synthesis of cartilage-specific matrix and collagen, further improving its ability to repair defects. The application of HAT resulted in an initial burst release of HA, which degraded slowly . Finally, HAT combined with microfracture-inducing bone marrow stem cells could significantly improve the cartilage microenvironment and regeneration of cartilage defects. Our results indicate that HA is a suitable material for developing growth factor carriers, whereas HAT is a promising candidate for cartilage regeneration. Furthermore, this differentiated strategy provides a rapid and effective clinical approach for next-generation cartilage regeneration.
113Growth factor-loaded sulfated microislands in granular hydrogels promote hMSCs migration and chondrogenic differentiation.PubMed
Anna Puiggalí-Jou, Maryam Asadikorayem, Katharina Maniura-Weber, et al.
Acta Biomater. 2023 Aug;166:69-84. doi: 10.1016/j.actbio.2023.03.045. Epub 2023 Apr 6.
Cell-based therapies for articular cartilage lesions are expensive and time-consuming; clearly, a one-step procedure to induce endogenous repair would have significant clinical benefits. Acellular heterogeneous granular hydrogels were explored for their injectability, cell-friendly cross-linking, and ability to promote migration, as well as to serve as a scaffold for depositing cartilage extracellular matrix. The hydrogels were prepared by mechanical sizing of bulk methacrylated hyaluronic acid (HAMA) and bulk HAMA incorporating sulfated HAMA (SHAMA). SHAMA's negative charges allowed for the retention of positively charged growth factors (GFs) (e.g., TGFB3 and PDGF-BB). Mixtures of HAMA and GF-loaded SHAMA microgels were annealed by enzymatic cross-linking, forming heterogeneous granular hydrogels with GF deposits. The addition of GF loaded sulfated microislands guided cell migration and enhanced chondrogenesis. Granular heterogeneous hydrogels showed increased matrix deposition and cartilage tissue maturation compared to bulk or homogeneous granular hydrogels. This advanced material provides an ideal 3D environment for guiding cell migration and differentiation into cartilage. STATEMENT OF SIGNIFICANCE: Acellular materials which promote regeneration are of great interest for repair of cartilage defects, and they are more cost- and time-effective compared to current cell-based therapies. Here we develop an injectable, granular hydrogel system which promotes cell migration from the surrounding tissue, facilitating endogenous repair. The hydrogel architecture and chemistry were optimized to increase cell migration and extracellular matrix deposition. The present study provides quantitative data on the effect of microgel size and chemical modification on cell migration, growth factor retention and tissue maturation.
114Three-Dimensional Bioprinting of Growth Differentiation Factor 5-Preconditioned Mesenchymal Stem Cell-Derived Exosomes Facilitates Articular Cartilage Endogenous Regeneration.PubMed
Yazhe Zheng, Liwei Fu, Zhichao Zhang, et al.
ACS Nano. 2025 Apr 29;19(16):15281-15301. doi: 10.1021/acsnano.4c13492. Epub 2025 Apr 17.
The repair of articular cartilage defects remains a major regenerative and clinical challenge. Exosomes (Exos) derived from mesenchymal stem cells (MSCs) have good application potential in cartilage tissue engineering. Numerous studies have indicated that appropriate preconditioning methods can promote the therapeutic effect of Exos. Growth differentiation factor 5 (GDF-5) plays a critical role in chondrogenesis and regeneration. In this study, GDF-5 was used to precondition synovial mesenchymal stem cells (SMSCs) to increase the chondrogenic-promoting effect of Exos (G-Exos). In addition, we demonstrated that G-Exos rich in miR-383-3p increased the chondrogenic potential of SMSCs by activating the Kdm2a/SOX2 signaling pathway. On this basis, G-Exos were loaded into a glycyrrhizic acid/methacrylate-acylated hyaluronic acid (GA/HA/G-Exos) scaffold via digital light processing (DLP) bioprinting to maintain bioactivity and sustained release. The GA/HA/G-Exos scaffolds not only presented significant biological properties in vitro but also significantly promoted the remodeling of the joint cavity regenerative microenvironment and the regeneration of articular cartilage in Sprague-Dawley rats. This study provides a promising cell-free regenerative strategy for cartilage defect repair via the use of engineered exofunctionalized biological scaffolds.
115Chondrogenic and chondroprotective response of composite collagen I/II-hyaluronic acid scaffolds within an inflammatory osteoarthritic environment.PubMed
Carly M Battistoni, Javier Munoz Briones, Douglas K Brubaker, et al.
Biomater Sci. 2025 Jun 10;13(12):3252-3263. doi: 10.1039/d5bm00033e.
Inflammation plays a key role in cartilage damage that occurs in osteoarthritis (OA). However, assessments of tissue-engineered constructs for cartilage regeneration generally do not consider their performance in the presence of inflammation. In this work, the chondrogenic differentiation potential of mesenchymal stromal cells (MSCs) was evaluated in the presence of both chondrogenic factors and inflammatory cytokines, and cartilage formation, degradative response, and inflammatory response were characterized. The addition of cytokines reduced cartilage production, increased cell proliferation, and resulted in an increase in inflammatory markers. Incorporation of hyaluronic acid (HA) had little impact on both collagen fibril microstructure and mechanical properties, two gel properties known to affect cell response, and thus allows the work to probe the biological impact of HA without the confounding effect of these gel properties. Regardless of environment, HA did not change cartilage production. The inflammatory response was similar with or without HA in terms of IL-6 and IL-10 secretion whereas IL-8 production exhibited some correlation with HA concentration as observed a linear regression model. Additionally, in the presence of cytokines, inclusion of HA statistically decreased the gene- and protein-level expression of matrix metalloproteinase-13 (MMP-13). Thus, when exposed to both chondrogenic growth factors and inflammatory cytokines within a chondrogenic-promoting collagen I/II blended hydrogel, chondrogenic differentiation of MSCs was limited by the inflammatory environment. These findings emphasize the importance of understanding how biomaterials affect cell responses within disease-relevant inflammatory environments.
116Naked mole-rat hyaluronan.PubMed
Francisco Alejandro Lagunas-Rangel
Biochimie. 2024 May;220:58-66. doi: 10.1016/j.biochi.2023.12.008. Epub 2023 Dec 27.
Naked mole rats (NMRs) are renowned for their exceptional longevity and remarkable maintenance of health throughout their lifetime. Their subterranean lifestyle has led to adaptations that have resulted in elevated levels of a very high molecular weight hyaluronan in their tissues. Hyaluronan, a glycosaminoglycan, is a key component of the extracellular matrix, which plays a critical role in maintaining tissue structure and regulating cell signaling pathways. This phenomenon in NMRs is attributed to a higher processing and production capacity by some of their hyaluronan synthases, along with lower degradation by certain hyaluronidases. Furthermore, this adaptation indirectly confers several advantages to NMRs, such as the preservation of skin elasticity and youthful appearance, accelerated wound healing, protection against oxidative stress, and resistance to conditions such as cancer and arthritis, largely attributable to CD44 signaling and other intricate mechanisms. Thus, the main objective of this study was to conduct a comprehensive study of the distinctive features of NMR hyaluronan, particularly emphasizing the currently known molecular mechanisms that contribute to its beneficial properties. Furthermore, this research delves into the potential applications of NMR hyaluronan in both cosmetic and therapeutic fields, as well as the challenges involved.
117Increased hyaluronan by naked mole-rat Has2 improves healthspan in mice.PubMed
Zhihui Zhang, Xiao Tian, J Yuyang Lu, et al.
Nature. 2023 Sep;621(7977):196-205. doi: 10.1038/s41586-023-06463-0. Epub 2023 Aug 23.
Abundant high-molecular-mass hyaluronic acid (HMM-HA) contributes to cancer resistance and possibly to the longevity of the longest-lived rodent-the naked mole-rat. To study whether the benefits of HMM-HA could be transferred to other animal species, we generated a transgenic mouse overexpressing naked mole-rat hyaluronic acid synthase 2 gene (nmrHas2). nmrHas2 mice showed an increase in hyaluronan levels in several tissues, and a lower incidence of spontaneous and induced cancer, extended lifespan and improved healthspan. The transcriptome signature of nmrHas2 mice shifted towards that of longer-lived species. The most notable change observed in nmrHas2 mice was attenuated inflammation across multiple tissues. HMM-HA reduced inflammation through several pathways, including a direct immunoregulatory effect on immune cells, protection from oxidative stress and improved gut barrier function during ageing. These beneficial effects were conferred by HMM-HA and were not specific to the nmrHas2 gene. These findings demonstrate that the longevity mechanism that evolved in the naked mole-rat can be exported to other species, and open new paths for using HMM-HA to improve lifespan and healthspan.
118A Trickster in Disguise: Hyaluronan's Ambivalent Roles in the Matrix.PubMed
Lena Bohaumilitzky, Ann-Kathrin Huber, Eva Maria Stork, et al.
Front Oncol. 2017 Oct 9;7:242. doi: 10.3389/fonc.2017.00242. eCollection 2017.
Hyaluronan (HA) is a simple but diverse glycosaminoglycan. It plays a major role in aging, cellular senescence, cancer, and tissue homeostasis. In which way HA affects the surrounding tissues greatly depends on the molecular weight of HA. Whereas high molecular weight HA is associated with homeostasis and protective effects, HA fragments tend to be linked to the pathologic state. Furthermore, the interaction of HA with its binding partners, the hyaladherins, such as CD44, is essential for sustaining tissue integrity and is likewise related to cancer. The naked mole rat, a rodent species, possesses a special form of very high molecular weight (vHMW) HA, which is associated with the extraordinary cancer resistance and longevity of those animals. This review addresses HA and its diverse facets: from HA synthesis to degradation, from oligomeric HA to vHMW-HA and from its beneficial properties to the involvement in pathologies. We further discuss the functions of HA in the naked mole rat and compare them to human conditions. Though intensively researched, this simple polymer bears some secrets that may hold the key for a better understanding of cellular processes and the development of diseases, such as cancer.
119Hyaluronan goes to great length.PubMed
Vera Gorbunova, Masaki Takasugi, Andrei Seluanov
Cell Stress. 2020 Jul 17;4(9):227-229. doi: 10.15698/cst2020.09.231.
Hyaluronan is a major non-protein component of extracellular matrix that affects biomechanical properties of tissues and interacts with cell receptors. Hyaluronan is a linear glycosaminoglycan composed of repeating disaccharides of (β, 1-4)-glucuronic acid (GlcUA) and (β, 1-3)-N-acetyl glucosamine (GlcNAc). The length of hyaluronan can range from an oligomer to an extremely long form up to millions of daltons. The concept that emerged in the field is that high (HMW-HA) and low (LMW-HA) molecular weight hyaluronans have different biological properties and trigger different signaling cascades within the cells. LMW-HA is associated with inflammation, tissue injury and metastasis, while HMW-HA improves tissue homeostasis and has anti-inflammatory and antimetastatic properties. HMW-HA is used in the clinic to treat arthritis, and as a filler in surgery and in the form of rinses to treat local inflammation. However, HMW-HA products used in the clinic come in a range of sizes between 0.5-6 mDa that are used interchangeably. Remarkably, the tissues of a long-lived and cancer-resistant rodent, the naked mole rat, contain abundant HA of very high molecular weight. While human fibroblasts secrete HA up to 2 MDa, naked mole rat fibroblasts produce HA of 6-12 MDa. Does this very high HMW-HA (vHMW-HA) differ functionally from HMW-HA? We found that vHMW-HA has superior cytoprotective properties compared to HMW-HA, and interacts differently with the CD44 receptor leading to distinct transcriptional changes (Takasugi (2020), Nat Commun). These results indicate that vHMW-HA has greater therapeutic benefits than the standard HMW-HA.
120Abundance and size of hyaluronan in naked mole-rat tissues and plasma.PubMed
Delphine Del Marmol, Susanne Holtze, Nadia Kichler, et al.
Sci Rep. 2021 Apr 12;11(1):7951. doi: 10.1038/s41598-021-86967-9.
Large amounts of ultra-high molecular weight hyaluronan (HA) have been described as the main cause of cancer resistance in naked mole-rats (Heterocephalus glaber, NMR). Our work examined HA metabolism in these rodents more closely. HA was localized and quantified using HA binding proteins. Its molecular weight was determined using size exclusion chromatography and gel electrophoresis, HA family gene expression using RNAseq analysis, and hyaluronidase activity using zymography. Guinea pigs (Cavia porcellus) and mice (Mus musculus) were used as controls for some experiments. We found that HA localization was similar in NMR, guinea pig, and mouse tissues but NMR had larger amounts and higher molecular weight (maximum, around 2.5 MDa) of HA in serum and almost all tissues tested. We could not find ultra-high molecular weight HA (≥ 4 MDa) in NMR samples, in contrast to previous descriptions. Hyaluronidase-1 had lower expression and activity in NMR than mouse lymph nodes. RNAseq results showed that, among HA family genes, Tnfaip6 and hyaluronidase-3 (Hyal3) were systematically overexpressed in NMR tissues. In conclusion, NMR samples, contrary to expectations, do not harbor ultra-high molecular weight HA, although its amount and average molecular weight are higher in NMR than in guinea pig tissues and serum. Although hyaluronidase expression and activity are lower in NMR than mouse lymph nodes, this not sufficient to explain the presence of high molecular weight HA. A different activity of the NMR HA synthases remains possible. These characteristics, together with extremely high Hyal3 and Tnfaip6 expression, may provide the NMR with a bespoke, and perhaps protective, HA metabolism.
121Evolution of High-Molecular-Mass Hyaluronic Acid is Associated with Subterranean Lifestyle.PubMed
Yang Zhao, Zhizhong Zheng, Zhihui Zhang, et al.
bioRxiv. 2023 May 9:2023.05.08.539764. doi: 10.1101/2023.05.08.539764.
Hyaluronic acid (HA) is a major component of extracellular matrix (ECM) which plays an important role in development, cellular response to injury and inflammation, cell migration, and cancer. The naked mole-rat (NMR, ) contains abundant high-molecular-mass HA (HMM-HA) in its tissues, which contributes to this species' cancer resistance and possibly longevity. Here we report that abundant HMM-HA is found in a wide range of subterranean mammalian species, but not in phylogenetically related aboveground species. These species accumulate abundant HMM-HA by regulating the expression of genes involved in HA degradation and synthesis and contain unique mutations in these genes. The abundant high molecular weight HA may benefit the adaptation to subterranean environment by increasing skin elasticity and protecting from oxidative stress due to hypoxic subterranean environment. HMM-HA may also be coopted to confer cancer resistance and longevity to subterranean mammals. Our work suggests that HMM-HA has evolved with subterranean lifestyle.
122Naked mole-rat TMEM2 lacks physiological hyaluronan-degrading activity.PubMed
Shinya Sato, Yukiko Mizutani, Minori Abe, et al.
Arch Biochem Biophys. 2024 Sep;759:110098. doi: 10.1016/j.abb.2024.110098. Epub 2024 Jul 14.
Mouse transmembrane protein 2 (mTMEM2) has been identified as a hyaluronidase, which has extracellularly G8 and GG domains and PbH1 repeats; however, our previously study showed that human TMEM2 (hTMEM2) is not a catalytic hyaluronidase due to the absence of the critical amino acid residues (His248/Ala303) in the GG domain. Naked mole-rats (NMRs) accumulate abundant high-molecular weight hyaluronan (HA) in their tissues, suggesting decreased HA degradation. Therefore, we aimed to evaluate the HA-degrading activity of NMR TMEM2 (nmrTMEM2) and compare it with those of mTMEM2 and hTMEM2. The amino acid residues of nmrTMEM2 (Asn247/Val302) are similar to Asn248/Phe303 of hTMEM2, and nmrTMEM2-expressing HEK293T cells showed negligible activity. We confirmed the significance of these amino acid residues using an inactive chimeric TMEM2 with the human GG domain, which acquired catalytic activity when Asn248/Phe303 was substituted with His248/Ala303. Semi-quantitative comparison of the activities of the membrane-fractions derived from m/h/nmrTMEM2-expressing HEK293T cells revealed that at least 20- and 14-fold higher amounts of nmr/hTMEM2 were required to degrade HA to the same extent as by mTMEM2. Thus, unlike mTMEM2, nmrTMEM2 is not a physiological hyaluronidase. The inability of nmrTMEM2 to degrade HA might partially account for the high-molecular-weight HA accumulation in NMR tissues.
123Cancer resistance, high molecular weight hyaluronic acid, and longevity.PubMed
Gary J Fisher
J Cell Commun Signal. 2015 Mar;9(1):91-2. doi: 10.1007/s12079-015-0278-6. Epub 2015 Mar 5.
Longevity varies greatly among mammals. The naked mole rat is among the longest-lived rodents, having an average lifespan of 32 years, compared to the similarly-sized house mouse with lifespan of 4 years. The rate of cancer also varies widely among mammals and interestingly, the naked mole rat is essentially cancer-free (Gorbunova et al., Nat Rev Genet 15(531):540, 2014). A series of elegant studies (Tian et al. Nature 499:346-349, 2013) has revealed that this cancer resistance derives from the abundant production of high molecular weight hyaluronic acid. Remarkably, high molecular weight hyaluronic acid, which accumulates within the extracellular matrix, stimulates an intracellular pathway that induces expression of p16(ink4a) and suppresses oncogenic transformation.
124Hyaluronic Acid and Its Synthases-Current Knowledge.PubMed
Klaudia Palenčárová, Romana Köszagová, Jozef Nahálka
Int J Mol Sci. 2025 Jul 22;26(15):7028. doi: 10.3390/ijms26157028.
Hyaluronic acid (HA) is a linear heteropolysaccharide that naturally occurs in vertebrates. Thanks to its unique physico-chemical properties, it is involved in many key processes in living organisms. These biological activities provide the basis for its broad applications in cosmetics, medicine, and the food industry. The molecular weight of HA might vary significantly, as it can be less than 10 kDa or reach more than 6000 kDa. There is a strong correlation between variations in its molecular weight and bioactivities, as well as with various pathological processes. Consequently, monodispersity is a crucial requirement for HA production, together with purity and safety. Common industrial approaches, such as extraction from animal sources and microbial fermentation, have limits in fulfilling these requests. Research and protein engineering with hyaluronic acid synthases can provide a strong tool for the production of monodisperse HA. One-pot multi-enzyme reactions that include in situ nucleotide phosphate regeneration systems might represent the future of HA production. In this review, we explore the current knowledge about HA, its production, hyaluronic synthases, the most recent stage of in vitro enzymatic synthesis research, and one-pot approaches.
125Microbial hyaluronic acid production.PubMed
Barrie Fong Chong, Lars M Blank, Richard Mclaughlin, et al.
Appl Microbiol Biotechnol. 2005 Jan;66(4):341-51. doi: 10.1007/s00253-004-1774-4. Epub 2004 Nov 13.
Hyaluronic acid (HA) is a commercially valuable medical biopolymer increasingly produced through microbial fermentation. Viscosity limits product yield and the focus of research and development has been on improving the key quality parameters, purity and molecular weight. Traditional strain and process optimisation has yielded significant improvements, but appears to have reached a limit. Metabolic engineering is providing new opportunities and HA produced in a heterologous host is about to enter the market. In order to realise the full potential of metabolic engineering, however, greater understanding of the mechanisms underlying chain termination is required.
126Tapping on the Potential of Hyaluronic Acid: from Production to Application.PubMed
Priya Shukla, Rupika Sinha, Shubhankar Anand, et al.
Appl Biochem Biotechnol. 2023 Nov;195(11):7132-7157. doi: 10.1007/s12010-023-04461-6. Epub 2023 Mar 24.
The manufacture, purification, and applications of hyaluronic acid (HA) are discussed in this article. Concerning the growing need for affordable, high-quality HA, it is essential to consider diverse production techniques using renewable resources that pose little risk of cross-contamination. Many microorganisms can now be used to produce HA without limiting the availability of raw materials and in an environmentally friendly manner. The production of HA has been associated with Streptococci A and C, explicitly S. zooepidemicus and S. equi. Different fermentation techniques, including the continuous, batch, fed-batch, and repeated batch culture, have been explored to increase the formation of HA, particularly from S. zooepidemicus. The topic of current interest also involves a complex broth rich in metabolites and residual substrates, intensifying downstream processes to achieve high recovery rates and purity. Although there are already established methods for commercial HA production, the anticipated growth in trade and the diversification of application opportunities necessitate the development of new procedures to produce HA with escalated productivity, specified molecular weights, and purity. In this report, we have enacted the advancement of HA technical research by analyzing bacterial biomanufacturing elements, upstream and downstream methodologies, and commercial-scale HA scenarios.
127Insight into hyaluronic acid molecular weight control.PubMed
Esteban Marcellin, Jennifer A Steen, Lars K Nielsen
Appl Microbiol Biotechnol. 2014 Aug;98(16):6947-56. doi: 10.1007/s00253-014-5853-x. Epub 2014 Jun 24.
Hyaluronic acid (HA) is a ubiquitous polysaccharide found in humans, animals, bacteria, algae and molluscs. Simple yet sophisticated, HA demonstrates unique and valuable rheological properties. In solution, HA behaves as a stiffened random coil and the resultant behaviour, even at low concentrations, is far from Newtonian or 'ideal'. These rheological properties are heavily influenced by molecular weight (MW), so it is not surprising that many of the biological functions of HA are dependent on molecular size. The current billion dollar market for HA continues to grow rapidly, both in gross production and the number of applications for its use. Increasing demand, in conjunction with a reticence to use animal-derived HA, has revitalised the market for HA produced by bacterial fermentation. Although the genes and pathways involved in bacterial production of HA are well characterised, the mechanisms that underlie HA MW control are less well understood. By performing a thorough analysis of the proposed mechanisms of MW control in bacterial fermentation, this mini-review tries to elucidate the challenges and future directions for bacterial HA biosynthesis.
128Efficient production of high-molecular-weight hyaluronic acid with a two-stage fermentation.PubMed
J Liu, Y Wang, Z Li, et al.
RSC Adv. 2018 Oct 24;8(63):36167-36171. doi: 10.1039/c8ra07349j. eCollection 2018 Oct 22.
In this paper, the production pattern of hyaluronic acid (HA) was revealed: the chain growth of HA mainly occurred in the first half of fermentation while the product accumulated throughout the fermentation period. Attempts were made to develop a two-stage fermentation process which provided high-level synthesis in both product titer and molecular weight. The pH was kept at 8.0 with a temperature of 31 °C in the first fermentation stage (0-10 h) to promote the growth of weight-average molecular weight ( ), and the pH and temperature were maintained at 7.0 and 37 °C, respectively, in the following fermentation stage to facilitate HA accumulation. In addition, constant rates of aeration (1 vvm) and agitation (600 rpm) were adopted. The two-stage fermentation provided a balanced result in which a product titer of 4.75 g L and a of 2.36 × 10 Da were achieved under optimized conditions. The process introduces an effective way to produce HA considering the effect of segmented control strategy.
129The rate and evenness of the substitutions on hyaluronan grafted by dodecanoic acid influenced by the mixed-solvent composition.PubMed
Eva Kutálková, Josef Hrnčiřík, Roman Witasek, et al.
Int J Biol Macromol. 2021 Oct 31;189:826-836. doi: 10.1016/j.ijbiomac.2021.08.137. Epub 2021 Aug 21.
In this work, low molecular weight (17 kDa) hyaluronan was modified by dodecanoyl substituents. The activation of dodecanoic acid was mediated by benzoyl chloride towards the preparation of a mixed anhydride, which reacts in a second step with HA in water mixed with an organic solvent. The effect of the cosolvent was studied and showed an even distribution of substituents and higher reaction rate in water: 1,4-dioxane compared to water:tert-butanol where substituents occupy adjacent positions. The chemical characterization of the prepared derivatives was elucidated by NMR, FTIR spectroscopy, thermal analyses, and gas chromatography, while the distribution of substituents was evaluated by enzymatic degradation. Molecular-dynamics simulations reveal opposite solvent separations around HA and dodecanoyl chains, that is stronger in water:tert-butanol solution. The resulting incompatibility of solvation-shells of the two entities repels the reaction intermediates from the HA chain and drives them towards the already bound substituents, explaining the observed differences in the distribution evenness. Thus, the influence of the solvent on the reaction selectivity is observed by shielding reactive sites around HA. Therefore, a control of the distribution of the substituents was obtained by defining the concentration of HA and used cosolvent.
130Repetitive Synthesis of High-Molecular-Weight Hyaluronic Acid with Immobilized Enzyme Cascades.PubMed
Johannes Gottschalk, Miriam Aßmann, Jürgen Kuballa, et al.
ChemSusChem. 2022 May 6;15(9):e202101071. doi: 10.1002/cssc.202101071. Epub 2021 Jul 12.
Industrial hyaluronic acid (HA) production comprises either fermentation with Streptococcus strains or extraction from rooster combs. The hard-to-control product quality is an obstacle to these processes. Enzymatic syntheses of HA were developed to produce high-molecular-weight HA with low dispersity. To facilitate enzyme recovery and biocatalyst re-use, here the immobilization of cascade enzymes onto magnetic beads was used for the synthesis of uridine-5'-diphosphate-α-d-N-acetyl-glucosamine (UDP-GlcNAc), UDP-glucuronic acid (UDP-GlcA), and HA. The combination of six enzymes in the UDP-sugar cascades with integrated adenosine-5'-triphosphate-regeneration reached yields between 60 and 100 % for 5 repetitive batches, proving the productivity. Immobilized HA synthase from Pasteurella multocida produced HA in repetitive batches for three days. Combining all seven immobilized enzymes in a one-pot synthesis, HA production was demonstrated for three days with a HA concentration of up to 0.37 g L , an average M of 2.7-3.6 MDa, and a dispersity of 1.02-1.03.
131Tailored molecular weight hyaluronic acid production by engineered Lactococcus lactis.PubMed
Sharath Soundiraraj, Nakul Ravishankar, Pandeeswari Jeeva, et al.
Microb Cell Fact. 2026 Feb 24;25(1):64. doi: 10.1186/s12934-026-02945-8.
BACKGROUND: Hyaluronic acid (HA) is a glycosaminoglycan with a wide range of biological functions that depend on its molecular weight (MW). Recently, there has been an increasing interest in producing HA at particular MWs for various cosmetic and biomedical applications. HA is traditionally produced by extraction or microbial fermentation, which is then subjected to chemical or enzymatic treatments to customize the MW. On the other hand, direct microbial synthesis at desired MWs has considerable advantages over conventional techniques. The present study introduces a combinatorial approach using four critical variables which influence the molecular weight of HA (MW): (1) Expression of HA synthases from different species (,,, and ) which intrinsically produce different MW; (2) Supply of HA precursors by varying heterlogous gene expression in the HA-precursor pathways ( vs. ); (3) Re-routing of metabolic fluxes by deletion of the lactate dehydrogenase () gene; and (4) Varying the initial glucose concentration in batch fermentation. RESULTS: Recombinant strains expressing HA synthase genes taken from diverse Streptococcal sp. were found to produce varying MW under otherwise identical genetic and bioreactor conditions. The HA synthases sourced from and synthesized higher MW, whereas those from produced lower MW. In silico analysis of the HA synthase sequences indicated that differences in the transmembrane regions among the various isoforms are the probable cause of variations in MW. Compared to their wild-type counterparts, -knockout strains showed a noticeable increase in MW due to a substantial increase in HA precursor levels. Further, the co-expression of in addition to , considerably increased MW due to a better balance of the intracellular HA-precursor ratios. This multiplexing approach, involving simultaneous manipulation of the above factors, allowed us to produce HA with tailored MW over a broad range from 0.2 to 2.6 MDa. CONCLUSION: Our technology eliminates the need for enzymatic desizing or post-processing of HA to achieve the desired MW. In summary, this multiplexing approach enables one-pot synthesis of desired MW, opening up new avenues for producing customized HA. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-026-02945-8.
132Independent control of molecular weight, concentration, and stiffness of hyaluronic acid hydrogels.PubMed
Jakob M Townsend, Megan E Sanders, Emi A Kiyotake, et al.
Biomed Mater. 2022 Sep 15;17(6). doi: 10.1088/1748-605X/ac8e41.
Hyaluronic acid (HA) hydrogels have been used for a multitude of applications, perhaps most notably for tissue engineering and regenerative medicine, owing to the versatility of the polymer and its tunable nature. Various groups have investigated the impact of hydrogel parameters (e.g. molecular weight, concentration, stiffness, etc)andto achieve desired material performance characteristics. A limitation in the literature to date has been that altering one hydrogel parameter (a 'manipulated variable') to achieve a given hydrogel characteristic (a 'controlled variable') changes two variables at a time (e.g. altering molecular weight and/or concentration to investigate cell response to stiffness). Therefore, if cell responses differ, it may be possible that more than one variable caused the changes in observed responses. In the current study, we leveraged thiol-ene click chemistry with a crosslinker to develop a method that minimizes material performance changes and permitted multiple material properties to be independently held constant to evaluate a single variable at a time. Independent control was accomplished by tuning the concentration of crosslinker to achieve an effectively constant stiffness for different HA hydrogel molecular weights and polymer concentrations. Specific formulations were thereby identified that enabled the molecular weight (76-1550 kDa), concentration (2%-10%), or stiffness (∼1-350 kPa) to be varied while the other two were held constant, a key technical achievement. The response of rat mesenchymal stem cells to varying molecular weight, concentration, and stiffness demonstrated consistent upregulation of osteocalcin gene expression. The methodology presented to achieve independent control of hydrogel parameters may potentially be adopted by others for alternative hydrogel polymers, cell types, or cell culture medium compositions to minimize confounding variables in experimental hydrogel designs.
133Magnesium metal-organic framework microneedles loaded with curcumin for accelerating oral ulcer healing.PubMed
Junhui Liu, Zhipeng Zhang, Xiufei Lin, et al.
J Nanobiotechnology. 2024 Sep 30;22(1):594. doi: 10.1186/s12951-024-02873-y.
Oral ulcers are a common oral mucosal disease that seriously affect the quality of life. Traditional drug treatments have shown unsatisfactory efficacy and potential adverse reactions. In this study, curcumin-loaded multifunctional magnesium metal-organic framework-embedded hyaluronic acid-soluble microneedles patches were developed to optimize treatment strategies for oral ulcers. This microneedles patch achieves efficient release of curcumin and Mg in the ulcer through precisely targeted delivery and controllable release mechanism, significantly regulates inflammation, promotes cell migration and angiogenesis, and accelerates the ulcer healing process. At the same time, the synergistic effect of curcumin and gallic acid effectively alleviated oxidative stress, while the backplate ε-poly-L-lysine and needle tip Mg jointly constructed an antibacterial barrier to effectively inhibit pathogens. Verification using an oral ulcer rat model showed that the microneedles patch exhibited excellent therapeutic effects. This not only opens up a new avenue for clinical oral treatment but also marks a breakthrough in nanobiomaterials science and drug delivery technology and heralds a broad prospect in the field of oral ulcer treatment in the future.
134Carboxymethyl chitosan microspheres loaded hyaluronic acid/gelatin hydrogels for controlled drug delivery and the treatment of inflammatory bowel disease.PubMed
Shangwen Zhang, Li Kang, Sheng Hu, et al.
Int J Biol Macromol. 2021 Jan 15;167:1598-1612. doi: 10.1016/j.ijbiomac.2020.11.117. Epub 2020 Nov 18.
A major drawback of oral treatment of inflammatory bowel disease (IBD) is the non-specific distribution of drugs during long-term treatment. Despite its effectiveness as an anti-inflammatory drug, curcumin (CUR) is limited by its low bioavailability in IBD treatment. Herein, a pH-sensitive composite hyaluronic acid/gelatin (HA/GE) hydrogel drug delivery system containing carboxymethyl chitosan (CC) microspheres loaded with CUR was fabricated for IBD treatment. The composition and structure of the composite system were optimized and the physicochemical properties were characterized using infrared spectroscopy, X-ray diffraction, swelling, and release behavior studies. In vitro, the formulation exhibited good sustained release property and the drug release rate was 65% for 50 h. In vivo pharmacokinetic experiments indicated that high level of CUR was maintained in the colon tissue for more than 24 h; it also played an anti-inflammatory role by evaluating the histopathological changes through hematoxylin and eosin (H&E), myeloperoxidase (MPO), and immunofluorescent staining. Additionally, the formulation substantially inhibited the level of the main pro-inflammatory cytokines of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) secreted by macrophages, compared to the control group. The pharmacodynamic experiment showed that the formulation group of CUR@gels had the best therapeutic effect on colitis in mice. The composite gel delivery system has potential for the effective delivery of CUR in the treatment of colitis. This study also provides a reference for the design and preparation of a new oral drug delivery system with controlled release behavior.
135Plant-derived exosomes: Unveiling the similarities and disparities between conventional extract and innovative form.PubMed
Xintong Han, Wenying Zheng, Zijun Sun, et al.
Phytomedicine. 2025 Sep;145:157087. doi: 10.1016/j.phymed.2025.157087. Epub 2025 Jul 17.
BACKGROUND: Herbal medicine, historically valued for its multi-component therapeutic synergy, faces significant challenges in clinical translation due to crude raw materials, compositional complexity, and low bioavailability, as well as unclear effective substances and action targets. As an innovative form of herbal medicine, plant-derived exosomes (PDEs) exhibit their therapeutic potential from a unique phospholipid bilayer structure, which encapsulates bioactive molecules including proteins, lipids, RNAs, and metabolites. PURPOSE AND METHODS: This study aims to elucidate the components, bioavailability, drug delivery, and stability of PDEs by reviewing the keywords including exosome, nanoparticle, and vesicle regarding plants and herbs in the databases (e.g., PubMed, Web of Science, MEDLINE, and Google Scholar), specially focusing on the high-frequency medicinal species (e.g., Ginseng, Ginger, Pueraria root, Turmeric, and Wolfberry). RESULTS: Unlike conventional extracts of herbal medicine, which lose critical compounds during drying or alcohol-based processing, PDEs derived from fresh plant tissues preserve native phytochemical integrity and enhance bioactivity. Advanced extraction techniques and cargo-loading strategies further amplify their therapeutic potential, enabling precise modulation of pathways such as macrophage polarization and immune checkpoint. Despite these advantages, hurdles in standardizing isolation protocols, ensuring storage stability, and validating clinical efficacy remain key obstacles to widespread adoption. The natural surface proteins of PDEs, which facilitate receptor-specific targeting without synthetic modification, exemplify their ability to bridge the synergy of herbal medicine with precision medicine. CONCLUSIONS: These advances position PDEs as transformative agents of herbal medicine in chronic disease management, offering a paradigm shift toward stable, targeted, and efficacious phytotherapy.
136Plant Derived Exosome-Like Nanoparticles and Their Therapeutic Applications in Glucolipid Metabolism Diseases.PubMed
Ruolan Liu, Feng Zhang, Xiaoyun He, et al.
J Agric Food Chem. 2025 Mar 19;73(11):6385-6399. doi: 10.1021/acs.jafc.4c12480. Epub 2025 Mar 6.
Plant derived exosome-like nanoparticles (PELNs) are membrane structures isolated from different plants, which encapsulate many active substances such as proteins, lipids, and nucleic acids, which exert a substantial influence on many physiological processes such as plant growth and development, self-defense, and tissue repair. Compared with synthetic nanoparticles and mammalian cell derived exosomes (MDEs), PELNs have lower toxicity and immunogenicity and possess excellent biocompatibility. The intrinsic properties of PELNs establish a robust basis for their applications in the therapeutic management of a diverse array of pathologies. It is worth mentioning that PELNs have good biological targeting, which promotes them to load and deliver drugs to specific tissues, offering a superior development pathway for the construction of a new drug delivery system (DDS). Glucose and lipid metabolism is a vital life process for the body's energy and material supply. The maintenance of homeostatic balance provides a fundamental basis for the body's ability to adjust to modifications in both its internal and external environment. Conversely, homeostatic imbalance can lead to a range of severe metabolic disorders. This work provides a comprehensive overview of the extraction and representation methods of PELNs, their transportation and storage characteristics, and their applications as therapeutic agents for direct treatment and as delivery vehicles to enhance nutrition and health. Additionally, it examines the therapeutic efficacy and practical applications of PELNs in addressing abnormalities in glucose and lipid metabolism. Finally, combined with the above contents, the paper summarizes and provides a conceptual framework for the better application of PELNs in clinical disease treatment.
137Research advances in smart responsive-hydrogel dressings with potential clinical diabetic wound healing properties.PubMed
Ying Chen, Xing Wang, Sheng Tao, et al.
Mil Med Res. 2023 Aug 23;10(1):37. doi: 10.1186/s40779-023-00473-9.
The treatment of chronic and non-healing wounds in diabetic patients remains a major medical problem. Recent reports have shown that hydrogel wound dressings might be an effective strategy for treating diabetic wounds due to their excellent hydrophilicity, good drug-loading ability and sustained drug release properties. As a typical example, hyaluronic acid dressing (Healoderm) has been demonstrated in clinical trials to improve wound-healing efficiency and healing rates for diabetic foot ulcers. However, the drug release and degradation behavior of clinically-used hydrogel wound dressings cannot be adjusted according to the wound microenvironment. Due to the intricacy of diabetic wounds, antibiotics and other medications are frequently combined with hydrogel dressings in clinical practice, although these medications are easily hindered by the hostile environment. In this case, scientists have created responsive-hydrogel dressings based on the microenvironment features of diabetic wounds (such as high glucose and low pH) or combined with external stimuli (such as light or magnetic field) to achieve controllable drug release, gel degradation, and microenvironment improvements in order to overcome these clinical issues. These responsive-hydrogel dressings are anticipated to play a significant role in diabetic therapeutic wound dressings. Here, we review recent advances on responsive-hydrogel dressings towards diabetic wound healing, with focus on hydrogel structure design, the principle of responsiveness, and the behavior of degradation. Last but not least, the advantages and limitations of these responsive-hydrogels in clinical applications will also be discussed. We hope that this review will contribute to furthering progress on hydrogels as an improved dressing for diabetic wound healing and practical clinical application.
138pH-Sensitive Hydrogels Fabricated with Hyaluronic Acid as a Polymer for Site-Specific Delivery of Mesalamine.PubMed
Huma Liaqat, Syed Faisal Badshah, Muhammad Usman Minhas, et al.
ACS Omega. 2024 Jun 21;9(26):28827-28840. doi: 10.1021/acsomega.4c03240. eCollection 2024 Jul 2.
Hydrogels with the main objective of releasing mesalamine (5-aminosalicylic acid) in the colon in a modified manner were formulated in the present work using a free-radical polymerization approach. Different ratios of hyaluronic acid were cross-linked with methacrylic and acrylic acids using methylenebis(acrylamide). The development of a new polymeric network and the successful loading of drug were revealed by Fourier transform infrared spectroscopy. Thermogravimetric analysis demonstrated that the hydrogel was more thermally stable than the pure polymer and drug. Scanning electron microscopy (SEM) revealed a rough and hard surface which was relatively suitable for efficient loading of drug and significant penetration of dissolution medium inside the polymeric system. Studies on swelling and drug release were conducted at 37 °C in acidic and basic conditions (pH 1.2, 4.5, 6.8, and 7.4, respectively). Significant swelling and drug release occurred at pH 7.4. Swelling, drug loading, drug release, and gel fraction of the hydrogels increased with increasing hyaluronic acid, methacrylic acid, and acrylic acid concentrations, while the sol fraction decreased. Results obtained from the toxicity study proved the formulated system to be safe for biological systems. The pH-sensitive hydrogels have the potential to be beneficial for colon targeting due to their pH sensitivity and biodegradability. Inflammatory bowel disease may respond better to hydrogel treatment as compared to conventional dosage forms. Specific amount of drug is released from hydrogels at specific intervals to maintain its therapeutic concentration at the required level.
139Hyaluronic acid methacryloyl-based co-delivery system for aflibercept and miR-21-3p antagomir: a dual-therapeutic approach for diabetic retinopathy.PubMed
Xisen Ni, Ya Zhao, Junya Zhu, et al.
J Control Release. 2025 Oct 10;386:114077. doi: 10.1016/j.jconrel.2025.114077. Epub 2025 Aug 5.
Diabetic retinopathy (DR) is a leading cause of vision loss characterized by concurrent retinal vascular dysfunction and neurodegeneration. While current therapies primarily target vascular dysfunction, they offer limited neuroprotective benefits. In this study, we developed a novel light-responsive hydrogel composed of hyaluronic acid methacryloyl (HAMA) for the co-delivery of aflibercept and miR-21-3p antagomir (HAMA@(Ab+M21A)). This dual-therapeutic strategy was designed to concurrently target retinal vascular dysfunction and neurodegeneration. Upon light exposure, HAMA hydrogel undergoes rapid in situ crosslinking, exhibiting excellent ocular biocompatibility and sustained drug release over 45 days, in parallel with controlled biodegradation. HAMA@(Ab+M21A) effectively inhibits VEGF-induced vascular dysfunction, suppresses reactive gliosis, and promotes retinal ganglion cell survival in vitro and in vivo. Collectively, this study demonstrates the therapeutic potential of HAMA@(Ab+M21A) as a dual-function strategy for DR, providing both anti-angiogenic and neuroprotective effects to impede disease progression.
140Clickable and smart drug delivery vehicles accelerate the healing of infected diabetic wounds.PubMed
Mingyan Deng, Ye Wu, Yan Ren, et al.
J Control Release. 2022 Oct;350:613-629. doi: 10.1016/j.jconrel.2022.08.053. Epub 2022 Sep 6.
In this study, an adipic acid dihydrazide (ADH)/ tannic acid (TA)-grafted hyaluronic acid (HA)-based multifunctional hydrogel was synthesized through a spontaneous amino-yne click reaction and used to promote the improved healing of infected diabetic wounds. This hydrogel exhibited a range of beneficial properties such as tunable gelation time, adjustable mechanical properties, pH-sensitive response characteristics, excellent injectability, the ability to readily adhere to tissue, and ultra-intimate contact capabilities. Following the encapsulation of ultrasmall Ag nanoclusters (AgNCs) and deferoxamine loaded polydopamine/ hollow mesoporous manganese dioxide (PHMD, PDA/H-mMnO@DFO) nanoparticles, the prepared hydrogel presented with robust antibacterial, anti-inflammatory, and pro-angiogenic properties and a desirable smart drug release profile. In this fabricated platform, PHMD was able to effectively alleviate localized oxidative stress and prolonged oxygen deprivation via the decomposition of endogenous HO to produce O. Further in vivo assays revealed that this hydrogel was capable of facilitating the healing of infected wounds through the sequential engagement of antibacterial, anti-inflammatory, and pro-angiogenic activities. Together, this synthesized clickable environmentally-responsive hydrogel offers great promise as a tool that can be applied to aid in the healing of chronically infected diabetic wounds and other inflammatory conditions.
141Properties and in vitro drug release of pH- and temperature-sensitive double cross-linked interpenetrating polymer network hydrogels based on hyaluronic acid/poly (N-isopropylacrylamide) for transdermal delivery of luteolin.PubMed
A Rang Kim, Sang Lae Lee, Soo Nam Park
Int J Biol Macromol. 2018 Oct 15;118(Pt A):731-740. doi: 10.1016/j.ijbiomac.2018.06.061. Epub 2018 Jun 27.
In this study, we prepared double cross-linked interpenetrating polymer network (IPN) hydrogels composed of temperature sensitive poly (N-isopropylacrylamide) (PNIPAM) and pH sensitive hyaluronic acid (HA) by radical polymerization and Michael addition. Their physicochemical properties for transdermal delivery of luteolin inhibiting the hyperproliferation of keratinocytes in psoriasis were investigated and drug release studies were performed. Double networks of HA/PNIPAM IPN hydrogel were identified through FT-IR and CNMR. By measuring the swelling ratios pH and temperature sensitivity were confirmed, and it was influenced by the content of a cross-linking agent. As a result of texture analysis and rheometry, a IPN hydrogel with 3% crosslinker content had the most adhesive and stable cross-linked network. Therefore, luteolin was loaded on this hydrogel. Its drug release behavior was determined at various temperatures and pH using several drug release kinetic models. As a result of skin permeation study, HA/PNIPAM IPN hydrogel effectively delivers luteolin to the epidermis and dermis. No toxicity was observed as a result of observing cytotoxicity of the hydrogel for application to the skin. In conclusion, IPN hydrogels can be developed as carriers of transdermal delivery system of luteolin for psoriasis skin relief.
142Physicochemical properties of pH-sensitive hydrogels based on hydroxyethyl cellulose-hyaluronic acid and for applications as transdermal delivery systems for skin lesions.PubMed
Soon Sik Kwon, Bong Ju Kong, Soo Nam Park
Eur J Pharm Biopharm. 2015 May;92:146-54. doi: 10.1016/j.ejpb.2015.02.025. Epub 2015 Mar 6.
We investigated the physicochemical properties of pH-sensitive hydroxyethyl cellulose (HEC)/hyaluronic acid (HA) complex hydrogels containing isoliquiritigenin (ILTG), and discussed potential applications as transdermal delivery systems for the treatment of skin lesions caused by pH imbalance. HA has skin compatibility and pH functional groups and HEC serves as scaffold to build hydrogels with varied HCE:HA mass ratio. Hydrogels were synthesized via chemical cross-linking, and three-dimensional network structures were characterized via scanning electron microscopy (SEM). The swelling properties and polymer ratios of the hydrogels were investigated at pH values in the range 1-13. HECHA13 (i.e., an HEC:HA mass ratio of 1:3) was found to have optimal rheological and adhesive properties, and was used to investigate the drug release efficiency as a function of pH; the efficiency was greater than 70% at pH 7. Antimicrobial activity assays against Propionibacterium acnes were conducted to take advantage of the pH-sensitive properties of HECHA13. At pH 7, we found that HECHA13, which contained ILTG, inhibited the growth of P. acnes. Furthermore, HECHA13 was found to exhibit excellent permeability into the skin, which penetrated mostly via the hair follicle. These results indicate that this pH-sensitive hydrogel is effective as a transdermal delivery system for antimicrobial therapeutics, with potential applications in the treatment of acne.
143Artificial mucus layer formed in response to ROS for the oral treatment of inflammatory bowel disease.PubMed
Guangshuai Zhang, Dandan Song, Ruilong Ma, et al.
Sci Adv. 2024 Jul 26;10(30):eado8222. doi: 10.1126/sciadv.ado8222.
The artificial mucus layer, such as hydrogels, used to repair the damaged intestinal barrier, is a promising treatment for inflammatory bowel disease (IBD). However, the currently reported hydrogel-based artificial barriers are administered via rectal injection, causing unnecessary discomfort to patients. Herein, we report an oral hydrogel precursor solution based on thiol-modified hyaluronic acid (HASH). Owing to the reactive oxygen species (ROS)-responsive gelling behavior, our precursor solution formed an artificial mucus coating over the inflamed regions of the intestines, blocking microbial invasion and reducing abnormally activated immune responses. Notably, HASH also modulated the gut microbiota, including increasing the diversity and enhancing the abundance of short-chain fatty acid-associated bacteria, which play a key role in gut homeostasis. We believe that the ROS-responsive artificial mucus layer is a promising strategy for the oral treatment of IBD.
144Injectable and body temperature sensitive hydrogels based on chitosan and hyaluronic acid for pH sensitive drug release.PubMed
Wei Zhang, Xin Jin, Heng Li, et al.
Carbohydr Polym. 2018 Apr 15;186:82-90. doi: 10.1016/j.carbpol.2018.01.008. Epub 2018 Jan 3.
Hydrogels based on chitosan/hyaluronic acid/β-sodium glycerophosphate demonstrate injectability, body temperature sensitivity, pH sensitive drug release and adhesion to cancer cell. The drug (doxorubicin) loaded hydrogel precursor solutions are injectable and turn to hydrogels when the temperature is increased to body temperature. The acidic condition (pH 4.00) can trigger the release of drug and the cancer cell (Hela) can adhere to the surface of the hydrogels, which will be beneficial for tumor site-specific administration of drug. The mechanical strength, the gelation temperature, and the drug release behavior can be tuned by varying hyaluronic acid content. The mechanisms were characterized using dynamic mechanical analysis, Fourier transform infrared spectroscopy, scanning electron microscopy and fluorescence microscopy. The carboxyl group in hyaluronic acid can form the hydrogen bondings with the protonated amine in chitosan, which promotes the increase of mechanical strength of the hydrogels and depresses the initial burst release of drug from the hydrogel.
145Platinum-Coordinated Dual-Responsive Nanogels for Universal Drug Delivery and Combination Cancer Therapy.PubMed
Qiu-Yi Duan, Ya-Xuan Zhu, Hao-Ran Jia, et al.
Small. 2022 Nov;18(46):e2203260. doi: 10.1002/smll.202203260. Epub 2022 Nov 4.
Developing a universal nanoplatform for efficient delivery of various drugs to target sites is urgent for overcoming various biological barriers and realizing combinational cancer treatment. Nanogels, with the advantages of both hydrogels and nanoparticles, may hold potential for addressing the above issue. Here, a dual-responsive nanogel platform (HPC nanogel) is constructed using β-cyclodextrin-conjugated hyaluronic acid (HA-βCD), polyethyleneimine (PEI), and cisplatin. HA-βCD and PEI compose the skeleton of the nanogel, and cisplatin molecules provide the junctions inside the skeleton, thus affording a multiple interactions-based nanogel. Besides, HA endows the nanogel with hyaluronidase (HAase)-responsiveness, and cisplatin guarantees the glutathione (GSH)-responsive ability, which make the nanogel a dual-responsive platform that can degrade and release the loaded drugs when encountering HAase or GSH. Additionally, the HPC nanogel possesses excellent small-molecule drug and protein loading and intracellular delivery capabilities. Especially, for proteins, their intracellular delivery via nanogels is not hindered by serum proteins, and the enzymes delivered into cells still maintain their catalytic activities. Furthermore, the nanogel can codeliver different cargoes to achieve "cocktail" chemotherapeutic efficacy and realize combination cancer therapy. Overall, the HPC nanogel can serve as a multifunctional platform capable of delivering desired drugs to treat cancer or other diseases.
146Hyaluronic Acid-Based Reactive Oxygen Species-Responsive Multifunctional Injectable Hydrogel Platform Accelerating Diabetic Wound Healing.PubMed
Chen Shi, Ying Zhang, Guanfu Wu, et al.
Adv Healthc Mater. 2024 Feb;13(4):e2302626. doi: 10.1002/adhm.202302626. Epub 2023 Nov 20.
Diabetic wounds are more likely to develop into complex and severe chronic wounds. The objective of this study is to develop and assess a reactive oxygen species (ROS)-responsive multifunctional injectable hydrogel for the purpose of diabetic wound healing. A multifunctional hydrogel (HA@Cur@Ag) is successfully synthesized with dual antioxidant, antibacterial, and anti-inflammatory properties by crosslinking thiol hyaluronic acid (SH-HA) and disulfide-bonded hyperbranched polyethylene glycol (HB-PBHE) through Michael addition; while, incorporating curcumin liposomes and silver nanoparticles (AgNPs). The HA@Cur@Ag hydrogel exhibits favorable biocompatibility, degradability, and injectivity. The outcomes of in vitro and in vivo experiments demonstrate that the hydrogel can effectively be loaded with and release curcumin liposomes, as well as silver ions, thereby facilitating diabetic wound healing through multiple mechanisms, including ROS scavenging, bactericidal activity, anti-inflammatory effects, and the promotion of angiogenesis. Transcriptome sequencing reveals that the HA@Cur@Ag hydrogel effectively suppresses the activation of the tumour necrosis factor (TNF)/nuclear factor κB (NF-κB) pathway to ameliorate oxidative stress and inflammation in diabetic wounds. These findings suggest that this ROS-responsive multifunctional injectable hydrogel, which possesses the ability to precisely coordinate and integrate intricate biological and molecular processes involved in wound healing, exhibits notable potential for expediting diabetic wound healing.
147ROS-responsive hydrogel for bone regeneration: Controlled dimethyl fumarate release to reduce inflammation and enhance osteogenesis.PubMed
Qiuxia Huang, Yang Qu, Mengchen Tang, et al.
Acta Biomater. 2025 Mar 15;195:183-200. doi: 10.1016/j.actbio.2025.02.026. Epub 2025 Feb 14.
Large bone defects, often arising from trauma or infection, pose a considerable therapeutic challenge due to their limited capacity for spontaneous healing, thus requiring bone graft materials for effective reparative procedures. The persistence of inflammation and elevated levels of reactive oxygen species (ROS) within these defect sites significantly impede bone regeneration process. Addressing this, an injectable hydrogel system with ROS-responsive functionality is developed, specifically tailored to the high ROS microenvironment characteristic of bone defects. This system incorporates hyaluronic acid functionalized with dopamine to introduce catechol moieties, and employs 4-formylphenylboronic acid as a crosslinking agent to form a dynamic hydrogel matrix (HAC) with carboxymethyl chitosan. The HAC hydrogel serves as a carrier for dimethyl fumarate (DMF), a compound with established anti-inflammatory and antioxidant effects, enabling its controlled release in response to ROS levels. Herein, we investigated the physicochemical properties of DMF loaded hydrogel (DHAC) by microstructure observation, in vitro degradation assay, self-healing test, injectability experiments, DMF drug release assay. Meanwhile, we systematically investigated its effects on inflammation, intracellular ROS, and osteogenesis. Consequently, the DHAC significantly reduced pro-inflammatory cytokines secreted by RAW264.7 cells and scavenged intracellular ROS in MC3T3 cells. This effect was accompanied by an augmentation in the osteogenic potential of MC3T3 cells and a promotion in the repair of cranial defects in rats. The DHAC, which exhibits anti-inflammatory, antioxidant, and osteogenic activity, hold great potential as an effective strategy for the management of large bone defects. STATEMENT OF SIGNIFICANCE: Here, a novel dimethyl fumarate-loaded ROS-responsive hydrogel system was developed for effective treatment of large bone defects. Our findings demonstrated that the hydrogel not only promotes bone regeneration but also controls inflammation, addressing two critical challenges in bone healing. Comprehensive evaluations show significant improvements in bone formation and reduction of pro-inflammatory cytokines in animal models. Additionally, the hydrogel exhibits excellent reactive oxygen species scavenging ability, effectively modulating oxidative stress in the bone defect microenvironment. Findings suggest the hydrogel system may serve as a promising therapeutic strategy for clinical management of critical-sized bone defects.
148Final report of the safety assessment of hyaluronic acid, potassium hyaluronate, and sodium hyaluronate.PubMed
Lillian C Becker, Wilma F Bergfeld, Donald V Belsito, et al.
Int J Toxicol. 2009 Jul-Aug;28(4 Suppl):5-67. doi: 10.1177/1091581809337738.
Hyaluronic acid, sodium hyaluronate, and potassium hyaluronate function in cosmetics as skin conditioning agents at concentrations up to 2%. Hyaluronic acid, primarily obtained from bacterial fermentation and rooster combs, does penetrate to the dermis. Hyaluronic acid was not toxic in a wide range of acute animal toxicity studies, over several species and with different exposure routes. Hyaluronic acid was not immunogenic, nor was it a sensitizer in animal studies. Hyaluronic acid was not a reproductive or developmental toxicant. Hyaluronic acid was not genotoxic. Hyaluronic acid likely does not play a causal role in cancer metastasis; rather, increased expression of hyaluronic acid genes may be a consequence of metastatic growth. Widespread clinical use of hyaluronic acid, primarily by injection, has been free of significant adverse reactions. Hyaluronic acid and its sodium and potassium salts are considered safe for use in cosmetics as described in the safety assessment.
149Effectiveness of topical hyaluronic acid of different molecular weights in xerosis cutis treatment in elderly: a double-blind, randomized controlled trial.PubMed
Parikesit Muhammad, Endi Novianto, Mirawati Setyorini, et al.
Arch Dermatol Res. 2024 Jun 3;316(6):329. doi: 10.1007/s00403-024-03003-2.
Dry skin is a common dermatological condition that frequently affects the elderly. A contributing cause to dry skin is a reduced concentration of hyaluronic acid (HA) in both the epidermis and dermis. The effectiveness of moisturizer containing HA as a therapy for dry skin is impacted by its specific molecular weight. Low molecular weight HA (LMWHA) is believed to be more effective in replenishing skin hydration in aging skin compared to High Molecular Weight HA (HMWHA) due to its ability to penetrate the stratum corneum. However, there is a lack of clinical research supporting this claim. A double-blind, randomized controlled trial was conducted on 36 residents of a nursing home in Jakarta. The participants, aged between 60 and 80 years, had been diagnosed with dry skin. Each test subject was administered three distinct, randomized moisturizing lotions (LMWHA, HMWHA, or vehicle), to be topically applied to three separate sites on the leg. Skin capacitance (SCap), transepidermal water loss (TEWL), and specified symptom sum score (SRRC) were measured at weeks 0, 2, and 4. After four weeks of therapy, area that was treated with LMWHA showed greater SCap values compared to the area treated with HMWHA (56.37 AU vs. 52.37 AU, p = 0.004) and vehicle (56.37 AU vs. 49.01 AU, p < 0.001). All groups did not show any significant differences in TEWL and SRRC scores. No side effects were found in all groups. The application of a moisturizer containing LMWHA to the dry skin of elderly resulted in significant improvements in skin hydration compared to moisturizers containing HMWHA and vehicle. Furthermore, these moisturizers demonstrated similar safety in treating dry skin in the elderly. ClinicalTrials.gov Identifier NCT06178367, https://clinicaltrials.gov/study/NCT06178367 .
150Hyaluronic Acid in Topical Applications: The Various Forms and Biological Effects of a Hero Molecule in the Cosmetics Industry.PubMed
Catherine Zanchetta, Amandine Scandolera, Romain Reynaud
Biomolecules. 2025 Nov 26;15(12):1656. doi: 10.3390/biom15121656.
BACKGROUND: Hyaluronan, or hyaluronic acid (HA), is a glycosaminoglycan with structural and signaling functions playing key roles in human skin homeostasis. It ensures hydration and biomechanical properties of this tissue as well as regulates cell adhesion, migration, proliferation, and inflammation. Its biocompatibility, viscoelastic properties, biological functions, and large-scale sustainable bioproduction made this polysaccharide a hero molecule of the cosmetic industry. METHODS: A literature search was conducted to discuss the skin and hair benefits of the external use of HA and its derivatives. Four main questions were addressed: What are the different forms of HA in cosmetic formulations? What about their safety? Does HA penetrate human skin and hair? What are the benefits and mode of actions of HA, and its derivatives, in the fields of cosmetic and dermatology? RESULTS: The analysis revealed HA below 100 kDa to penetrate skin, and lower molecular weight being able to reach the dermis. The safety of HA-containing formulations has been evaluated in several clinical trials and is supported by independent reports of commercial ingredients. We described HA molecules having beneficial effects on skin and hair, as well as their mode of action. CONCLUSIONS: This review provides comprehensive information on the nature and efficacy of topical HA, and its derivatives, in cosmetic applications, with an emphasis on hair care. New areas of research were highlighted as the vectorization of high-molecular-weight HA.
151Safety and Efficacy of a "High and Low Molecular Weight Hyaluronic Acid Hybrid Complex" Injection for Face Rejuvenation.PubMed
Taraneh Yazdanparast, Azin Ayatollahi, Aniseh Samadi, et al.
J Cosmet Dermatol. 2025 Apr;24(4):e70117. doi: 10.1111/jocd.70117.
INTRODUCTION: Hyaluronic acid (HA)-based formulations could have remarkable efficacy in treating a wide range of skin defects, including skin aging. The purpose of the study was the evaluation of the clinical safety and efficacy of a high and low molecular weight HA hybrid complex injection for skin rejuvenation. METHOD: In this single-arm, before-and-after clinical study, 20 subjects with wrinkled, dry, or rough skin were enrolled. They received two treatment sessions, each of 2 mL of stabilized high- and low-molecular-weight HA through intradermal injections in five bioaesthetic points with an interval of 4 weeks. Efficacy assessment measures included biophysical and sonographic parameters and the Global Aesthetic Improvement Scale (GAIS) score. Pain assessment, safety profile, and subject satisfaction were also reported. RESULTS: A significant improvement in skin firmness was demonstrated in both follow-up visits. The Transepidermal water loss (TEWL) and the dermis ecodensity improved significantly in the first follow-up visit. A statistically significant increase in the dermis thickness was seen in the second follow-up visit. The median GAIS score indicated an average improvement of 51%-75%. The median overall satisfaction score was 7 and 6 in the first and second follow-up visits. No important side effects were observed. The average pain VAS score was 2 out of 10. CONCLUSION: This new HA-based formulation is a safe and efficient treatment option to restore the vitality and turgidity of the skin. TRIAL REGISTRATION: ClinicalTrials.gov identifier: IRCT20150101020514N17 with ethics code IR.TUMS.TIPS.REC.1401.083.
152Role and Effectiveness of Intra-articular Injection of Hyaluronic Acid in the Treatment of Knee Osteoarthritis: A Systematic Review.PubMed
Sumant Chavda, Syed Arman Rabbani, Tarun Wadhwa
Cureus. 2022 Apr 26;14(4):e24503. doi: 10.7759/cureus.24503. eCollection 2022 Apr.
Osteoarthritis (OA) is a degenerative joint disease that causes persistent joint pain and stiffness of mainly the large peripheral weight-bearing joints. It is a leading cause of functional disability and poor quality of life. Various modalities of therapy are recommended by different research organizations at different stages of OA including non-pharmacological, pharmacological, and surgical interventions. Intra-articular injections of hyaluronic acid (HA) is widely used for over three decades in the treatment of OA. However controversies exist regarding its safety and efficacy, the number of injections and courses, type of preparation, duration of its effects, and combining it with other drugs or molecules. This study aimed to review the most recent data available in the published literature to address these. Electronic databases like Medline, Embase, ProQuest, and Google Scholar were searched for articles using keywords, intraarticular injections, hyaluronic acid, and osteoarthritis knee. The review was carried out as per PRISMA guidelines. Thirty-eight randomized control trials (RCTs) investigating the efficacy and safety of intra-articular injection of HA were included in the systematic review. Out of the 38 studies, 22 (57.9%) were double-blind, eight (21%) single-blind, three (7.9%) non-blind, four (10%) with simple randomization, and one (2.7%) was open-labeled. Total 5,025 patients were included in these studies. The mean age of the patients was 60.28 years and the osteoarthritis grade of the knee joint was 1 to 3. HA was studied as a test preparation in 19 (50%) while in another 19 (50%) it was studied as a control. In 24 (63.2%) studies, HA was used as high molecular weight preparation in eight (21%) as low molecular weight preparation while in six studies the information was not available. HA was used as a standalone preparation in 31 studies, in two studies it was injected with platelet-rich plasma (PRP) and with either low-level laser therapy (LLLT), triamcinolone (TA), betamethasone (CS), poly deoxyribonucleotide (PDRN) or dexamethasone (DX) in one study each. In the majority of the studies, HA was given as a single injection (52.6% studies) or weekly three injections (28.9% studies). In 13.2 %, it was given as weekly 5 injections and in 5.3% as weekly two injections. IA-HA injections have a limited role in the treatment of knee osteoarthritis in those patients who do not have sufficient pain relief with topical or oral medication and physical therapy. It is safe and effective except for minor side effects such as local pain and swelling lasting for a few days. Severe allergic reactions are extremely rare. They provide adequate pain relief and functional improvement for up to six months irrespective of a number of injections and type of preparations used. The combination formulations with corticosteroids or PRP or MSCs show better results than HA alone. Combining HA with newer molecules such as peptides or diclofenac for sustained and disease-modifying effects requires more studies in the future.
153A Comprehensive Review of Viscosupplementation in Osteoarthritis of the Knee.PubMed
Jacquelin Peck, Annabel Slovek, Paulo Miro, et al.
Orthop Rev (Pavia). 2021 Jul 10;13(2):25549. doi: 10.52965/001c.25549. eCollection 2021.
PURPOSE OF REVIEW: The purpose of this systematic review is to discuss emerging evidence in the field of viscosupplementation for chronic knee pain secondary to Osteoarthritis (OA). This review focuses on types of viscosupplementation that are clinically available currently, evidence to support their use, contraindications, and adverse events. RECENT FINDINGS: OA, also known as degenerative joint disease, is the most common form of arthritis in the United States, affecting 54.4 million, or 22.7% of the adult population. The knee is the most common joint affected in OA, with up to 41% involvement, 30% in the hands, and 19% in the hips. The pathophysiology of OA is complex, with contributing factors including mechanical stress to the joint, as well as many person-specific factors such as genetic susceptibility, ethnicity, nutrition, and sex. Treatment modalities include weight control, exercise, non-steroidal and steroidal anti-inflammatory drugs, opioids, intra-articular platelet-rich plasma, placebo, corticosteroid injection, intra-articular viscosupplementation, and surgery. Viscosupplementation consists of injection of hyaluronic acid (HA) into affected joints, intending to restore the physiologic viscoelasticity in the synovial fluid (SF) in the absence of inflammation. HA has also been shown to downregulate pro-inflammatory factors, such as PGE2 and NFkB, and proteases and proteinases known to break down the joint matrix.The contraindications for HA injection are similar to any other injection therapy, and adverse events are usually mild, local, and transient. Viscosupplementation (VS) is effective over placebo and more effective than NSAIDs and corticosteroids in pain reduction and improved functionality; however, guidelines recommend neither for nor against its use, demonstrating variability in the existing evidence base.Current VS options divide primarily into native vs. cross-linked and low-molecular-weight vs. high-molecular-weight. Current treatment options include Hylan g-f-20, Sodium Hyaluronate preparations (Suparts Fx, Euflexxa, Gelsyn-3, Durolane, Hyalgen), single-use agents (Gel-One, Synvisc-One, Monovisc), and Hyaluronan (Orthovisc, Monovisc, Hymovic). They share a common safety profile, and all have evidence supporting their efficacy. Their specific details are reviewed here. SUMMARY: OA is the most common form of arthritis. It is a chronic, debilitating illness with a high impact on the functionality and quality of life of a significant part of the population in the western world. Treatments include medical management, physical therapy, activity modification, injection, and surgery. VS effectively reduces pain, increases functionality, and delays surgery in the knee to treat osteoarthritis. While previous studies have demonstrated variable results, more evidence is becoming available generally supportive of the benefit of VS in the treatment of knee OA.
154Differentiating factors of intra-articular injectables have a meaningful impact on knee osteoarthritis outcomes: a network meta-analysis.PubMed
Mark Phillips, Christopher Vannabouathong, Tahira Devji, et al.
Knee Surg Sports Traumatol Arthrosc. 2020 Sep;28(9):3031-3039. doi: 10.1007/s00167-019-05763-1. Epub 2020 Jan 3.
PURPOSE: There are a number of developments in intra-articular therapies that have been determined to be differentiating factors within the classes of treatments. This study evaluated the efficacy and safety of intra-articular treatments of primary knee osteoarthritis in the short term (3 months follow-up), using a network meta-analysis design, while taking within-class differentiating factors into consideration. METHODS: A literature search of MEDLINE (through OVID), EMBASE (through OVID), Cochrane Central Register of Controlled Trials for all trials comparing intra-articular therapies was conducted on November 12, 2018. The treatments assessed were high molecular weight and low molecular weight hyaluronic acid injections, extended-release corticosteroids, standard-release corticosteroids, platelet-rich plasma, and saline. A frequentist network meta-analysis was conducted for each outcome. RESULTS: Sixty-four articles (9710 patients) met the inclusion criteria. High molecular weight hyaluronic acid (- 0.53, 95% CI - 0.81 to - 0.25) and PRP (- 0.79, 95% CI - 1.32 to - 0.26) were the only treatments with a confidence interval that lay completely above the MID threshold; however, PRP results varied within sensitivity analyses. For the function analysis, high molecular weight hyaluronic acid (SMD - 0.76, 95% CI - 1.30 to - 0.22) was the only treatment with a confidence interval entirely above the MID. Extended-release corticosteroid demonstrated a possible benefit in functional improvement (SMD - 0.98, 95% CI - 1.79 to - 0.17) compared to that of standard-release corticosteroid (SMD - 0.14, 95% CI - 0.72 to 0.44). CONCLUSION: High molecular weight HA was the only treatment to surpass the MID for both pain and function outcomes. Extended-release corticosteroids may provide additional clinical benefit over standard-release corticosteroids. Platelet-rich plasma demonstrated possibly beneficial results; however, wide confidence intervals and sensitivity analyses made the conclusions of efficacy uncertain. LEVEL OF EVIDENCE: Level 1. Systematic review of level 1 evidence.
155Intraarticular Hyaluronic Acid Preparations for Knee Osteoarthritis: Are Some Better Than Others?PubMed
David Webner, Yili Huang, Charles D Hummer
Cartilage. 2021 Dec;13(1_suppl):1619S-1636S. doi: 10.1177/19476035211017320. Epub 2021 May 28.
OBJECTIVE: This literature review summarizes evidence on the safety and efficacy of intraarticular hyaluronic acid (IAHA) preparations approved in the United States for the treatment of osteoarthritis of the knee. DESIGN: A systematic literature search was performed in PubMed, Ovid MEDLINE, and SCOPUS databases. Only studies in which clinical outcomes of individual IAHA preparations alone could be assessed when compared to placebo, no treatment, other standard knee osteoarthritis treatments, and IAHA head-to-head studies were selected. RESULTS: One hundred nine articles meeting our inclusion criteria were identified, including 59 randomized and 50 observational studies. Hylan G-F 20 has been the most extensively studied preparation, with consistent results confirming efficacy in placebo-controlled studies. Efficacy is also consistently reported for Supartz, Monovisc, and Euflexxa, but not for Hyalgan, Orthovisc, and Durolane. In the head-to-head trials, high-molecular-weight (MW) Hylan G-F 20 was consistently superior to low MW sodium hyaluronate preparations (Hyalgan, Supartz) up to 20 weeks, whereas one study reported that Durolane was noninferior to Supartz. Head-to-head trials comparing high versus medium MW preparations all used Hylan G-F 20 as the high MW preparation. Of the IAHA preparations with strong evidence of efficacy in placebo-controlled studies, Euflexxa was found to be noninferior to Hylan G-F 20. There are no direct comparisons to Monovisc. One additional IAHA preparation (ie, Synovial), which has not been assessed in placebo-controlled studies, was also noninferior to Hylan G-F 20. CONCLUSION: IAHA efficacy varies widely across preparations. High-quality studies are required to assess and compare the safety and efficacy of IAHA preparations.