Markov P A, Eremin P S, Paderin N M, Kostromina E Yu, Greben A I, Gilmutdinova I R
National Medical Research Centre for Rehabilitation and Balneology' of the Ministry of Health of Russia, Moscow, Russia.
Institute of Physiology of Federal Research Centre "Komi Science Centre of the Urals Branch of the Russian Academy of Sciences", Syktyvkar, Russia.
Dokl Biochem Biophys. 2024 Dec;519(1):540-546. doi: 10.1134/S1607672924701187. Epub 2024 Oct 31.
One of the key stages of wound healing is the phase of inflammation, which is a transitional process between hemostasis and wound healing. Each stage of the inflammatory-reparative process is characterized by its own value of the acidity of the wound bed. For example, in the acute stage of inflammation, the acidity of the medium in the wound bed decreases to pH 5.5-6. The chronic stage of the inflammatory process, on the contrary, is accompanied by an increase in pH to 8. To date, the effect of biomaterials containing components of the intercellular matrix of the human dermis on fibroblasts under acidosis and alkalosis has not been fully investigated.
The aim of this study was to characterize the effect of bioplastic material based on collagen, hyaluronic acid, and elastin on the viability and proliferative activity of human fibroblasts in conditions simulating the acidity of acute and chronic wounds.
Bioplastic material was made according to the method described in RF patent no. 2722744. Adhesive properties and proliferative activity of human fibroblasts were assessed visually using fluorescent microscopy. The number of apoptotic and necrotic cells was assessed by flow cytometry (BD FACSCanto II) using the commercial FITC Annexin V Apoptosis Detection Kit I (BD Pharmingen). The strength, Young's modulus, and elasticity of the gels were determined on a TA.XT-plus texture analyzer (Stable Micro Systems, Great Britain).
Using the methods of luminescent microscopy and flow cytometry, we found that the cell viability (namely, adhesive properties and proliferative activity) decreases after incubation on condition mimic of physiological acidosis. We found that bioplastic material contributes to the preservation of adhesive properties, viability, and proliferative activity of fibroblasts in physiological acidosis conditions. The results obtained indicate that bioplastic material based on soluble dermis components can be used as a biologically active component of wound dressings to increase the effectiveness of reparative regeneration, especially in cases of excessive acute inflammation.
伤口愈合的关键阶段之一是炎症期,它是止血和伤口愈合之间的过渡过程。炎症修复过程的每个阶段都以伤口床酸度的自身值为特征。例如,在炎症急性期,伤口床介质的酸度降至pH 5.5 - 6。相反,炎症过程的慢性期则伴随着pH值升高至8。迄今为止,含有人类真皮细胞间基质成分的生物材料在酸中毒和碱中毒情况下对成纤维细胞的影响尚未得到充分研究。
本研究的目的是在模拟急性和慢性伤口酸度的条件下,表征基于胶原蛋白、透明质酸和弹性蛋白的生物塑料材料对人类成纤维细胞活力和增殖活性的影响。
生物塑料材料按照俄罗斯联邦专利第2722744号所述方法制备。使用荧光显微镜通过视觉评估人类成纤维细胞的黏附特性和增殖活性。使用商业FITC Annexin V凋亡检测试剂盒I(BD Pharmingen)通过流式细胞仪(BD FACSCanto II)评估凋亡和坏死细胞的数量。在TA.XT-plus质地分析仪(英国稳定微系统公司)上测定凝胶的强度、杨氏模量和弹性。
使用发光显微镜和流式细胞术方法,我们发现,在模拟生理酸中毒条件下孵育后,细胞活力(即黏附特性和增殖活性)降低。我们发现生物塑料材料有助于在生理酸中毒条件下保持成纤维细胞的黏附特性、活力和增殖活性。所得结果表明,基于可溶性真皮成分的生物塑料材料可作为伤口敷料的生物活性成分,以提高修复再生的有效性,尤其是在急性炎症过度的情况下。