Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Tissue Engineering, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
J Biomater Appl. 2021 Nov;36(5):789-802. doi: 10.1177/08853282211021692. Epub 2021 Jun 1.
Horseradish peroxidase (HRP)-catalyzed hydrogels are considered to be an important platform for tissue engineering applications. In this study, we investigated the chondrogenic capacity of phenolated (1.2%) alginate-(0.5%) collagen hydrogel on human amniotic mesenchymal stem cells after 21 days. Using NMR, FTIR analyses, and SEM imaging, we studied the phenolation and structure of alginate-collagen hydrogel. For physicochemical evaluations, gelation time, mechanical properties, swelling, and degradation rate were assessed. The survival rate was monitored using the MTT assay and DAPI staining. Western blotting was performed to measure the chondrogenic differentiation of cells. NMR showed successful phenolation of the alginate-collagen hydrogel. FTIR exhibited the interaction between the functional groups of collagen with phenolated alginate. SEM showed the existence of collagen microfibrils in the alginate-collagen hydrogel. Compared to phenolated alginate, the addition of collagen increased hydrogel elasticity by 10%. Both swelling rate and biodegradability were reduced in the presence of collagen. We noted an increased survival rate in phenolated alginate-collagen compared to the control cells (p < 0.05). Western blotting revealed the increase of chondrocyte-associated proteins such as SOX9 and COL2A1 in phenolated-alginate-collagen hydrogels after 21 days. These data showed that phenolated alginate-collagen hydrogel is an appropriate 3 D substrate to induce chondrogenic capacity of human mesenchymal stem cells.
辣根过氧化物酶（HRP）催化水凝胶被认为是组织工程应用的重要平台。在这项研究中，我们研究了经过 21 天酚醛化（1.2%）海藻酸钠-（0.5%）胶原水凝胶对人羊膜间充质干细胞的软骨生成能力。我们使用 NMR、FTIR 分析和 SEM 成像研究了海藻酸钠-胶原水凝胶的酚醛化和结构。为了进行理化评估，评估了凝胶时间、机械性能、溶胀和降解率。通过 MTT 测定和 DAPI 染色监测存活率。通过 Western blot 测定测量细胞的软骨分化。NMR 显示海藻酸钠-胶原水凝胶的酚醛化成功。FTIR 显示了胶原蛋白的功能基团与酚醛化海藻酸钠之间的相互作用。SEM 显示了胶原微纤维在海藻酸钠-胶原水凝胶中的存在。与酚醛化海藻酸钠相比，胶原的添加使水凝胶弹性增加了 10%。在存在胶原的情况下，溶胀率和生物降解性降低。与对照细胞相比，我们注意到酚醛化海藻酸钠-胶原中的细胞存活率增加（p<0.05）。Western blot 显示，在 21 天后，酚醛化-海藻酸钠-胶原水凝胶中增加了软骨细胞相关蛋白，如 SOX9 和 COL2A1。这些数据表明，酚醛化海藻酸钠-胶原水凝胶是一种合适的 3D 基质，可以诱导人间充质干细胞的软骨生成能力。
