低强度高频振动对人牙周膜干细胞成骨分化能力的影响

Effect of low-magnitude high frequency vibration on osteogenic differentiation of human periodontal ligament stem cells

目的:研究低强度高频振动(low-magnitude high frequency vibration,LMHFV)对人牙周膜干细胞(human periodontal ligament stem cells,hPDLSCs)增殖、迁移、成骨分化能力的影响.方法:体外分离培养hPDLSCs;流式细胞术检测间充质干细胞表面标志物;加载LMHFV(加速度=0.3 g,频率=40 Hz,时间=15 min/24 h)刺激后,采用CCK-8试剂盒检测细胞增殖能力,通过细胞划痕实验检测细胞迁移能力;通过qRT-PCR、Western免疫印迹检测成骨相关基因、蛋白表达水平,通过茜素红染色检测细胞成骨分化能力.采用SPSS 21.0软件包对数据进行统计学分析.结果:加载振动刺激后,hPDLSCs的增殖能力增强,迁移能力上升;RUNX2、ALP、Col-1、OCN的mRNA表达量和蛋白表达量均上升,茜素红染色结果与qRT-PCR、Western免疫印迹结果一致.结论:LMHFV可提高hPDLSCs的增殖、迁移能力和成骨分化能力.

PURPOSE: To investigate the effect of low-magnitude high frequency vibration (LMHFV) on proliferation, migration ability and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). METHODS: hPDLSCs were isolated from premolar and randomized into vibration culture group (magnitude : 0.3 g;frequency: 40 Hz;time: 15 min/24 h) and static culture group. CCK-8 was used to identity the proliferation of hPDLSCs. Wound-healing assay was used to evaluate migration ability of hPDLSCs. The osteogenesis gene expression was analyzed by RT-PCR, and the osteogenesis protein expression was analyzed by Western blot. The osteogenesis differentiation capability was evaluated by alizarin red staining. The data were analyzed using SPSS 21.0 software package. RESULTS: After LMHFV, the proliferation and migration ability of hPDLSCs were increased. The expression level of RUNX2, ALP, Col-1, and OCN was significantly augmented under LMHFV. Alizarin red staining and Western blot proved the same trend. CONCLUSIONS: The results demonstrate that LMHFV can promote hPDLSCs proliferation, migration ability and osteogenic differentiation.