低强度脉冲超声对不同钛表面骨髓间充干细胞成骨分化的影响

摘要

Objective:To observe the effects of low intensity pulsed ultrasound(LIPUS) on the osteogenic differentiation of rat bone marrow mesenchymal stem cells(BMSCs) on titanium surface.Methods:BMSCs from Wistar rat bone marrow were respectively cultured on the flat titanium surface and the large grain blast acid etched(SLA) titanium surface,and induced by mineralization medium.Then,the cells were interfered by LIPUS and a control condition.Alkaline phosphatase(ALP) were quantitative determinated after 3 and 7 d mineralization induction respectively,ALP staining were observed after 14 d induction.Alizarin red staining were observed after 21 d mineralization induction.Osteogenic related protein and gene expressions were detected after mineralization induction.Results:ALP in culture medium of LIPUS group was higher than that of the control group after 3 d and 7 d mineralization induction(P＜0.05).LIPUS group showed stronger ALP staining and alizarin staining,and more mineralized nodules than control group.The expression of osteogenic related proteins,including Runx2,BMP2,OPN in LIPUS group increased.Osteogenic related genes expression,including ALP,Runx2,BMP2,OPN,OCN and Col-1 of the LIPUS group increased.Conclusion:The osteogenic differentiation of BMSCs on the fiat titanium surface or SLA titanium surface can be promoted by LIPUS.%目的:观察低强度脉冲超声(LIPUS)对大鼠骨髓间充质干细胞(BMSCs)在钛片表面成骨分化的影响.方法:将体外培养的BMSCs分别接种于光滑钛表面和大颗粒喷砂酸蚀(SLA)钛表面,进行矿化诱导并实施超声干预和假刺激.于矿化诱导后3、7d进行碱性磷酸酶(ALP)定量检测,14 d进行ALP染色观察;矿化诱导21 d后行茜素红染色;于矿化诱导14、21 d检测成骨相关基因及蛋白表达.结果:矿化诱导后3、7d超声组ALP活性较对照组高(P＜0.05),14 d超声组与对照组相比钛片表面ALP染色明显深染;茜素红染色显示,21 d后超声组的染色更明显且形成更多的矿化结节;超声组的成骨相关蛋白成骨转录因子2、骨形成蛋白、骨桥蛋白表达增高;RT-PCR检测到成骨相关基因、ALP、成骨转录因子2、骨形成蛋白、骨桥蛋白、骨钙素、Ⅰ型胶原蛋白均有不同程度表达增高(P＜0.05).结论:LIPUS可以促进光滑及SLA钛表面BMSCs的成骨分化.