Li Bao, Gao Yongli, Guo Likun, Fan Yujiang, Kawazoe Naoki, Fan Hongsong, Zhang Xingdong, Chen Guoping
National Engineering Research Center for Biomaterials, Sichuan University, 29 Wangjiang Road, Chengdu, Sichuan 610064, China.
Research Center for Functional Materials, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan.
Regen Biomater. 2018 Jun;5(3):159-166. doi: 10.1093/rb/rby009. Epub 2018 May 3.
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 .
通过将苯基叠氮基引入聚乙烯醇(PVA)中合成了光反应性聚乙烯醇(PRPVA),并将其应用于表面改性。PRPVA均匀地或以微图案形式接枝到细胞培养板表面。在细胞培养板表面和PVA改性表面上培养的人间充质干细胞(hMSCs)表现出不同的行为。细胞在细胞培养板表面上粘附并良好铺展,而它们在PVA接枝表面上根本不粘附。当hMSCs在PVA微图案化表面上培养时,它们形成了细胞微图案。在软骨诱导培养基中在PVA均匀改性表面上培养2周后,细胞形成小球。这些小球经苏木精/伊红、番红O/固绿和甲苯胺蓝阳性染色,并且它们也通过II型胶原蛋白和蛋白聚糖免疫组织化学染色染成棕色。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改性表面上培养的hMSCs进行软骨分化,并且有可能用PVA改性表面构建无支架的软骨样小球。