高糖微环境下EMD对钛表面BMSCs成骨分化的影响及其机制研究

The effects and the mechanism of EMD in the osteogenic differentiation of BMSCs on the titanium surface in a high glucose microenvironment

目的:探讨高糖微环境中釉基质衍生物(EMD)对喷砂酸蚀(SLA)钛表面上骨髓间充质干细胞(BMSCs)成骨分化与黏附的影响及机制。方法:BMSCs分别培养于正常糖(NG)和高糖(HG)微环境中,Transwell观察BMSCs的迁移情况;SLA钛表面培养BMSCs,使用ALP活性、茜素红染色半定量、扫描电镜、WB和RT-qPCR检测成骨和黏附的性能。结果:HG组细胞迁移、ALP活性、茜素红半定量和扫描电镜成骨分化能力减弱,WB和RT-qPCR检测成骨和黏附的蛋白和基因表达量均降低,添加EMD后改善。XAV-939处理后,Wnt/β-catenin信号通路下游的蛋白表达降低,EMD诱导后均增加。结论:高糖微环境下,EMD促进SLA钛表面BMSCs成骨分化和黏附,可能通过激活Wnt/β-catenin信号通路实现。

Objective:To investigate the effects and mechanism of enamel matrix derivatives(EMD)in the osteogenic differentiation and the adhesion of bone marrow mesenchymal stem cells(BMSCs)on the sandblasted acid-etched(SLA)titanium surface in a high glucose(HG)microenvironment.Methods:BMSCs were cultured in normal glucuse(NG)and HG microenvironment respectively,Transwell test was used to observe the migration of BMSCs.BMSCs were cultured on the SLA titanium surface.ALP activity test,alizarin red staining,SEM,WB and RT-qPCR were applied to detect the performance of osteogenesis and adhesion of the cells.Results:In HG group,cell migration,ALP activity and osteogenic differentiation ability were reduced,WB and RT-qPCR proved that the expression of osteogenes related proteins and genes decreased,but they were increased after EMD addition.The expression of downstream proteins of Wnt/β-catenin signaling pathway decreased after XAV-939 addition,and increased after EMD induction.Conclusion:Under HG microenvironment,EMD promotes the osteogenic differentiation and the adhesion of BMSCs on SLA titanium surface,possibly through activation of the Wnt/β-catenin signaling pathway.