生物活性玻璃陶瓷涂层与Hedgehog信号通路调控成骨  

Correlation of bioactive glass ceramic coating and Hedgehog signaling pathway in osteogenesis

作　　者：倪卫东[1] 方龙云[2] 

机构地区：[1]延边大学医学院,吉林省延吉市133002 [2]延边大学附属延边医院骨科,吉林省延吉市133002

出　　处：《中国组织工程研究》2016年第43期6424-6430,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81260279)~~

摘　　要：背景:生物活性玻璃陶瓷作为钛合金涂层的应用,已在临床上获得成功。而Hedgehog(Hh)信号通路与成骨关系密切,但目前为止还没有生物活性玻璃陶瓷涂层与Hh信号通路调控成骨过程的关系的研究。目的:探讨生物活性玻璃陶瓷涂层与Hh信号通路调控成骨过程的关系。方法:(1)制备生物活性玻璃陶瓷涂层材料,应用扫描及透射电子显微镜观察涂层结构;(2)将原代培养大鼠骨髓间充质干细胞接种在生物活性玻璃陶瓷涂层上,应用荧光定量RT-PCR、Western blot、免疫荧光染色等方法检测成骨标志物骨形态发生蛋白2和Hh信号通路关键因子Gli1(Gliloma-association oncogene homoglog)的表达,并观察细胞的迁移能力。结果与结论:(1)电镜下生物活性玻璃陶瓷涂层表面无裂纹,表面光滑,具有致密的介孔结构,涂层厚度约350 nm;(2)生物活性玻璃陶瓷组中骨形态发生蛋白2和Gli1 mR NA和蛋白表达水平显著高于对照组,且Gli1和骨形态发生蛋白2蛋白在成骨过程中相互联系;与对照组相比,生物活性玻璃陶瓷组的骨髓间充质干细胞的迁移能力明显增强。(3)结果提示生物活性玻璃陶瓷涂层通过增加Hh信号通路的转录因子Gli1的表达,进而促进激活Hh信号通路,增强Hh信号通路活性,协同增加骨形态发生蛋白2,共同促进成骨细胞的生长增殖。BACKGROUND： Bioactive glass ceramics （BGC） has been successfully applied as titanium alloy coating in the clinic. However, the correlation between Hedgehog signaling pathway and BGC coating in osteogenesis has not yet been reported. OBJECTIVE： To explore the correlation of Hedgehog signaling pathways and BGC coating in osteogenesis. METHODS： The BGC coating was prepared and observed by scanning and transmission electron microscopes. Primary cultured rat bone marrow mesenchymal stem cells were incubated onto the BGC coating. The expressions of bone morphogenetic protein 2 and Gliloma-association oncogene homoglog in the Hedgehog signaling pathway were detected using fluorescent quantitative RT-PCR, western blot assay, and immunofluorescence staining, and the cell miqration ability was observed. RESULTS AND CONCLUSION： Electron microscopes showed that there were no cracks on the smooth BGC coating that had the dense mesoporous structure, and the coating thickness was 350 nm. The mRNA and protein expressions of bone morphogenetic protein 2 and Gliloma-association oncogene homoglog in the BGC group were significantly higher than those in the control group; and the expresssed bone morphogenetic protein 2 and Gliloma-association oncogene homoglog proteins interacted with each other in the process of osteogenesis. The cell migration ability in the BGC group was obviously enhanced compared with the control group. These results indicate that the BGC coating increases the expression of Gliloma-association oncogene homoglog, then further activates the Hedgehog signaling pathway, and finally accelerates the osteoblast proliferation in combination with bone morphogenetic protein 2.

关 键 词：骨髓 间质干细胞 成骨细胞 组织工程 生物材料 骨生物材料 生物活性玻璃陶瓷 涂层 骨髓间充质干细胞 HEDGEHOG信号通路 GLI1 骨形态发生蛋白2 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]