胶原蛋白和20α羟胆固醇复合涂层对大鼠骨髓间充质干细胞成骨表达的影响

上海口腔医学 ›› 2016, Vol. 25 ›› Issue (3): 275-280.

胶原蛋白和20α羟胆固醇复合涂层对大鼠骨髓间充质干细胞成骨表达的影响

郑颖, 刘帆, 王岩, 刘奕

中国医科大学附属口腔医院奉天门诊,辽宁省口腔医学研究所, 辽宁沈阳 110013

收稿日期:2015-11-11 修回日期:2016-02-26 出版日期:2016-06-25 发布日期:2016-07-22
通讯作者: 刘奕,Tel（Fax）：024-31973999, E-mail: liuyi256@126.com E-mail:superzhengying@163.com
作者简介:郑颖（1985-）,女,硕士,医师
基金资助:
辽宁省公益研究团队建设项目（2015225090）; 辽宁省教育厅科学研究一般项目（L2015584）

Osteogenic expression of rat mesenchymal stem cell on collagen and 20α-hydroxycholesterol loaded on the apatite of titanium

ZHENG Ying, LIU Fan, WANG Yan, LIU Yi

Fengtian Dental Clinic, School of Stomatology, China Medical University. Shenyang 110013, Liaoning Province, China

Received:2015-11-11 Revised:2016-02-26 Online:2016-06-25 Published:2016-07-22

摘要/Abstract

摘要： 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。[关键词] 钛;磷灰石;胶原蛋白;羟胆固醇;成骨分化

关键词: Titanium, Apatite, Collagen, Hydroxycholesterol, Osteogenic expression

Abstract: PURPOSE: To investigate the effect of composite layers loaded with 20α-hydroxycholesterol and collagen on the surface of apatite-coated Ti substrates on osteogenesis of rat bone marrow mesenchymal stem cells (BMSCs). METHODS: BMSCs proliferation was quantitatively assessed by cell counting kit-8 (CCK-8). The early osteogenic expression of BMSCs on composite layers was evaluated by alkaline phosphatase (ALP) activity. The expression of osteocalcin (OC) and collagen TypeⅠ (Col Ⅰ) was evaluated by Western blot. The difference between the different groups was analyzed using SPSS 17.0 software package. RESULTS: CCK-8 results revealed that simultaneous loading of Col Ⅰ and HC on biomimetic apatite significantly promoted cell proliferation, while HC had no effect on cell proliferation. Dual loading of Col and HC onto apatite stimulated osteogenic differentiation in BMSCs, especially during the early stage. In addition, individual loading of HC seemed to be more efficient than collagen in promoting ALP activity in BMSCs. Dual loading of Col and HC synergistically promoted the expression of OC and ColⅠ, which were the marker of late osteogenic differentiation. HC had no positive effect on the expression of OC and Col Ⅰ. CONCLUSIONS: Dual loading of collagen and 20α-hydroxycholesterol on apatite-coated titanium substrates had favourable biocompatibility and synergistically promoted osteogenic expression.

Key words: Titanium, Apatite, Collagen, Hydroxycholesterol, Osteogenic expression

中图分类号:

R783.5

郑颖, 刘帆, 王岩, 刘奕. 胶原蛋白和20α羟胆固醇复合涂层对大鼠骨髓间充质干细胞成骨表达的影响[J]. 上海口腔医学, 2016, 25(3): 275-280.

ZHENG Ying, LIU Fan, WANG Yan, LIU Yi. Osteogenic expression of rat mesenchymal stem cell on collagen and 20α-hydroxycholesterol loaded on the apatite of titanium[J]. Shanghai Journal of Stomatology, 2016, 25(3): 275-280.

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