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基于硫酸软骨素和透明质酸的多空水凝胶支架,用于改善成骨和成软骨分化。

Chondroitin Sulfate and Hyaluronic Acid-Based PolyHIPE Scaffolds for Improved Osteogenesis and Chondrogenesis .

机构信息

Symbiosis Centre for Stem Cell Research (SCSCR), Symbiosis International (Deemed University), Pune 412115, India.

Symbiosis School of Biological Sciences (SSBS), Symbiosis International (Deemed University), Pune 412115, India.

出版信息

ACS Appl Bio Mater. 2024 Aug 19;7(8):5222-5236. doi: 10.1021/acsabm.4c00393. Epub 2024 Jul 15.

Abstract

Osteochondral damage, affecting the articular cartilage and the underlying subchondral bone, presents significant challenges in clinical treatment. Such defects, commonly seen in knee and ankle joints, vary from small localized lesions to larger defects. Current medical therapies encounter several challenges, such as donor shortages, drug side effects, high costs, and rejection problems, often resulting in only temporary relief. Highly porous emulsion-templated polymers (polyHIPEs) offer numerous potential benefits in the fabrication of scaffolds for tissue engineering and regenerative medicine. Polymeric scaffolds synthesized using a high internal phase emulsion (HIPE) technique, called PolyHIPEs, involve polymerizing a continuous phase surrounding a dispersed internal phase to form a solid, foam-like structure. A dense, porous design encourages cell ingrowth, nutrient delivery, and waste disposal from the scaffold, mimicking the cells' natural microenvironment. This study used hydroxyethyl methacrylate (HEMA) and acrylamide (AAM) polyHIPE scaffolds combined with extracellular matrix (ECM) components of the tissue, such as methacrylated hyaluronic acid (MHA) and methacrylated chondroitin sulfate (MCS), to prepare polyHIPE scaffolds. The mouse preosteoblast MC3T3-E1 cells and primary rat chondrocytes (harvested from male Wistar rats) were seeded on the scaffolds and cultured for 21 days to assess the osteogenesis and chondrogenesis . When compared to the AAM-MHA and AAM-MCS groups at day 21, scaffold groups HEMA-MHA and HEMA-MCS showed a significant rise in alkaline phosphatase (ALP) and calcium content. Chondrogenic markers such as glycosaminoglycan (GAG) and hydroxyproline were also assessed over a 21-day time point. On day 21, it was found that GAG and hydroxyproline production were considerably higher in the HEMA-MHA and HEMA-MCS scaffolds than in the AAM-MHA and AAM-MCS scaffolds. The overall studies showed that polyHIPE monolith scaffolds could favor cell adherence, survival ability, proliferation, differentiation, and ECM formation over 21 days. Thus, incorporating ECM components enhanced osteogenesis and chondrogenesis and can be further used as tissue repair models.

摘要

骨软骨损伤,影响关节软骨和其下的软骨下骨,在临床治疗中带来了重大挑战。这种损伤常见于膝关节和踝关节,从小的局限性病变到较大的损伤不等。目前的医学治疗方法存在许多挑战,如供体短缺、药物副作用、高成本和排斥问题,往往只能暂时缓解。高度多孔的乳液模板聚合物(polyHIPE)在组织工程和再生医学中支架的制造方面具有许多潜在的优势。使用高内相乳液(HIPE)技术合成的聚合物支架,称为 PolyHIPE,涉及聚合包围分散相的连续相以形成固体、泡沫状结构。致密、多孔的设计促进了细胞的向内生长、营养物质的输送和废物从支架中的处理,模拟了细胞的自然微环境。本研究使用羟乙基甲基丙烯酸酯(HEMA)和丙烯酰胺(AAM)polyHIPE 支架,并结合组织的细胞外基质(ECM)成分,如甲基丙烯酰化透明质酸(MHA)和甲基丙烯酰化硫酸软骨素(MCS),制备 polyHIPE 支架。将小鼠前成骨细胞 MC3T3-E1 细胞和原代大鼠软骨细胞(从雄性 Wistar 大鼠中收获)接种在支架上,并培养 21 天,以评估成骨和软骨形成。与第 21 天的 AAM-MHA 和 AAM-MCS 组相比,支架组 HEMA-MHA 和 HEMA-MCS 的碱性磷酸酶(ALP)和钙含量显著升高。还在 21 天的时间点评估了软骨形成标志物,如糖胺聚糖(GAG)和羟脯氨酸。在第 21 天,发现 HEMA-MHA 和 HEMA-MCS 支架中的 GAG 和羟脯氨酸产量明显高于 AAM-MHA 和 AAM-MCS 支架。总体研究表明,polyHIPE 整体支架在 21 天内可促进细胞黏附、存活能力、增殖、分化和 ECM 形成。因此,掺入 ECM 成分增强了成骨和软骨形成,并可进一步用作组织修复模型。

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