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目的:探讨固定在金属钛表面类骨磷灰石上的胶原蛋白(collagen,Col)和20α羟胆固醇(20α-hydroxycholesterol,HC)复合涂层对大鼠骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)成骨表达的影响。方法:通过CCK-8法评估BMSCs在固定了生物活性分子的钛表面上不同时间点的增殖能力;通过检测碱性磷酸酶活性(ALP activity)评估BMSCs在固定了生物活性分子的钛表面上不同时间点的早期分化能力;通过Western免疫印迹法检测BMSCs在不同时间点骨钙素(osteocalcin,OC)和I型胶原蛋白(collagen I,Col I)的表达。采用SPSS 17.0软件包对数据进行t检验。结果:CCK-8检测结果显示,共固定Col和HC的复合涂层提高了BMSCs的增殖能力,单一固定在具有类骨磷灰石钛表面上的HC对BMSCs的增殖能力无显著影响;检测ALP活性发现,共固定Col和HC的复合涂层显著提升了BMSCs的早期成骨分化能力,在单一固定HC涂层上,BMSCs的ALP活性比在单一固定Col的涂层高;Western免疫印迹结果显示,共固定Col和HC的复合涂层增加了BMSCs的OC和Col I表达,显著提高了BMSCs的晚期成骨能力;此外,在单一固定Col的涂层上,骨钙素和I型胶原蛋白的表达要高于在单一固定HC的涂层。结论:共固定的Col和HC复合涂层显示出良好的生物相容性,两者显著促进BMSCs的增殖能力和成骨分化能力。
OBJECTIVE: To investigate the effect of collagen, Col and 20α-hydroxycholesterol (HC) composite coating on bone mesenchymal stem cells (BMSCs) BMSCs) osteoblast expression. METHODS: BMSCs were evaluated for proliferation over time on titanium surfaces immobilized with bioactive molecules by CCK-8 assay; BMSCs were evaluated on titanium surfaces immobilized with bioactive molecules by detecting alkaline phosphatase activity (ALP activity) The differentiation ability of BMSCs at different time points was detected by Western blotting. The expression of osteocalcin (OC) and collagen I (Col I) at different time points were detected. Data was t-tested using SPSS 17.0 software package. Results: The results of CCK-8 showed that the co-immobilized Col and HC composite coatings increased the proliferation of BMSCs. HC immobilized on the surface of bone-like apatite titanium had no significant effect on the proliferation of BMSCs. The detection of ALP It was found that co-immobilization of Col and HC composite coatings significantly enhanced the early osteogenic differentiation of BMSCs. The ALP activity of BMSCs was higher than that of the single immobilized Col on a single fixed HC coating. Western blot results , Co-immobilization of Col and HC composite coatings increased the expression of OC and Col I in BMSCs, and significantly increased the late osteogenic potential of BMSCs. In addition, in the coating of a single fixed Col, osteocalcin and type I collagen The expression is higher than in a single fixed HC coating. CONCLUSION: Co-immobilized Col and HC composite coatings show good biocompatibility, both of which significantly promote the proliferation and osteogenic differentiation of BMSCs.