静态牵张力对人炎症牙周膜干细胞成骨分化及细胞骨架重组的影响

摘要

AIM:To investigate the effects of static mechanical strain(SMS) on the osteogenic differentiation potential and cytoskeletal reorganization of human periodontal ligament stem cells derived from chronic periodontitis tissue(PPDLSCs).METHODS:PPDLSCs were primarily cultured and identified.After being subjected to cyclic SMS with physiological loading of 12% at a frequency of 0.1 Hz for 12 h using a FX-4000T system,mineralization of PPDLSCs was examined by ALP staining,alizarin staining and ALP activity test.Cytoskeletal arrangement was observed by immunofluorescence staining and confocal microscopy.Protein expression was evaluated by Western blot analysis.RESULTS:SMS decreased the density of mineralized nodules and suppressed the cytoskeletal rearrangement of PPDLSCs,down-regulated ALP,RUNX2,ROCK1,cofilin and Rho-A protein expression,increased p-cofilin and Rho-GDI protein expression.CONCLUSION:SMS in the magnitude of 12% can suppress the osteogenic differentiation potential and cytoskeletal reorganization of PPDLSCs.%目的:探究静态牵张力(SMS)对人炎症来源的牙周膜干细胞(PPDLSCs)的成骨分化能力及细胞骨架重组的影响.方法:体外培养PPDLSCs,用FX-T4000力学加载系统对PPDLSCs体外施加力值为12%、频率为0.1 Hz的静态牵张力,加载12 h后(对照组不加力),分别通过ALP染色、茜素红染色检测其成骨分化能力;倒置显微镜观察细胞的形态变化;免疫荧光染色和共聚焦显微镜观察细胞骨架的改变;Western blot法检测ALP、RUNX2、ROCK1、p-cofilin、Rho-GDIα、Rho-A蛋白的表达水平. 结果: 实验组PPDLSCs的矿化结节形成量明显减少;细胞骨架弹力纤维的形成受到抑制;ALP、RUNX2、ROCK1、cofilin、Rho-A蛋白表达水平下降,p-cofilin、Rho-GDIα蛋白的表达水平则升高(P<0.05). 结论:12%静态牵张力抑制PPDLSCs成骨分化和细胞骨架的重组.