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大鼠间充质干细胞在负载于钛磷灰石上的胶原蛋白和20α-羟基胆固醇上的成骨表达

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

作者信息

Hao Wen-Xiu, Zheng Ying, Liu Fan, Wang Yan, Liu Yi

机构信息

Fengtian Dental Clinic, School of Stomatology, China Medical University. Shenyang 110013, Liaoning Province, China. E-mail:

出版信息

Shanghai Kou Qiang Yi Xue. 2016 Jun;25(3):275-80.

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.

摘要

目的

研究在磷灰石涂层钛基底表面负载20α-羟基胆固醇和胶原蛋白的复合层对大鼠骨髓间充质干细胞(BMSCs)成骨作用的影响。

方法

采用细胞计数试剂盒-8(CCK-8)定量评估BMSCs的增殖情况。通过碱性磷酸酶(ALP)活性评估BMSCs在复合层上的早期成骨表达。采用蛋白质免疫印迹法评估骨钙素(OC)和Ⅰ型胶原蛋白(Col Ⅰ)的表达。使用SPSS 17.0软件包分析不同组之间的差异。

结果

CCK-8结果显示,在仿生磷灰石上同时负载Col Ⅰ和HC可显著促进细胞增殖,而HC对细胞增殖无影响。在磷灰石上同时负载Col和HC可刺激BMSCs的成骨分化,尤其是在早期阶段。此外,单独负载HC在促进BMSCs的ALP活性方面似乎比胶原蛋白更有效。同时负载Col和HC可协同促进OC和ColⅠ的表达,这是晚期成骨分化的标志物。HC对OC和ColⅠ的表达没有积极作用。

结论

在磷灰石涂层钛基底上同时负载胶原蛋白和20α-羟基胆固醇具有良好的生物相容性,并协同促进成骨表达。

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