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Synthesis of photo-reactive poly (vinyl alcohol) and construction of scaffold-free cartilage like pellets in vitro

机译:光反应性聚乙烯醇的合成及体外无支架软骨样颗粒的构建

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Photo-reactive poly(vinyl alcohol) (PRPVA) was synthesized by introduction of phenyl azido groups into poly(vinyl alcohol) (PVA) and applied for surface modification. PRPVA was grafted onto cell culture plate surface homogeneously or in a micropattern. Human mesenchymal stem cells (hMSCs) cultured on cell culture plate surface and PVA-modified surface showed different behaviors. Cells adhered and spread well on cell culture plate surface, while they did not adhere on PVA-grafted surface at all. When hMSCs were cultured on PVA-micropatterned surface, they formed a cell micropattern. Cells formed pellets after cultured on PVA homogeneously modified surface in chondrogenic induction medium for 2 weeks. The pellets were positively stained by hematoxylin/eosin, safranin-O/fast green and toluidin blue, and they were also stained brown by Type II collagen and proteoglycan immunohistological staining. Real-time PCR analysis was conducted to investigate the expression of colI, colII, colX, aggrecan and sox9 mRNA. Results of gene expression were in agreement with those of histological and immunohistological observations. These results indicated that hMSCs cultured on PVA-modified surface performed chondrogenic differentiation, and it was possible to construct scaffold-free cartilage like pellets with PVA-modified surface in vitro.
机译:通过将苯基叠氮基引入聚乙烯醇(PVA)中来合成光反应性聚乙烯醇(PRPVA),并将其用于表面改性。将PRPVA均匀或以微模式移植到细胞培养板表面。在细胞培养板表面和PVA修饰的表面上培养的人间充质干细胞(hMSC)表现出不同的行为。细胞在细胞培养板表面上粘附并良好地扩散,而它们根本不粘附在PVA移植的表面上。当hMSCs在PVA微图案化的表面上培养时,它们形成了细胞微图案。在软骨诱导培养基中的PVA均质改性表面上培养2周后,细胞形成沉淀。用苏木精/曙红,番红O-O /坚牢绿和甲苯胺蓝对这些沉淀物进行阳性染色,并且通过II型胶原蛋白和蛋白聚糖免疫组织学染色将其染色为棕色。进行实时PCR分析以研究colI,colII,colX,聚集蛋白聚糖和sox9 mRNA的表达。基因表达结果与组织学和免疫组织学观察结果一致。这些结果表明,在PVA修饰的表面上培养的hMSC进行软骨形成分化,并且可以在体外构建具有PVA修饰的表面的无支架的软骨样颗粒。

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