【摘要】 目的探讨体外冲击波(extracorporeal shock wave,ESW)对大鼠骨间充质干细胞(mesenchymal stem cells,MSCs)成骨分化的影响及SDF-1/CXCR4通路在其中的作用。方法体外试验中分空白对照组(Ctrl组)、冲击波组(ESW组)及CXCR4特异性抑制剂AMD3100对照组(AMD组),采用最适宜能量ESW(500脉冲次数、10 k V)处理MSCs,通过ELISA检测SDF-1分泌表达,根据茜素红染色及碱性磷酸酶试验结果对比MSCs成骨分化的差别;体内试验则选取12周龄SD大鼠48只,随机分成Ctrl、ESW及AMD 3组各16只,采用右侧股骨中段骨缺损模型,各组骨缺损处植入载有该组细胞的PLGA支架,AMD组术后接受AMD3100注射(1 mg/kg/day),于术后第4、8周取材,通过Micro-CT、H-E染色及免疫组化评估骨缺损区域的骨愈合及新生骨形成情况。结果 ESW明显增加MSCs的SDF-1分泌量及体内外成骨分化作用,AMD3100部分抑制ESW促进作用。结论ESW可明显增加MSCs的成骨分化作用,通过分泌性蛋白SDF-1及CXCR4受体相互作用介导其促进骨缺损区域的新骨形成以促进骨折愈合,此研究为ESW在促进骨愈合临床治疗中的应用提供了理论基础。更多还原

【Abstract】 Objective To investigate the effect of extracorporeal shock wave( ESW) on osteogenic differentiation of rat bone mesenchymal stem cells( MSCs) and the role of SDF-1/CXCR4 pathway. Methods The MSCs were divided into 3 groups: the blank control group( Ctrl group),shock wave group( ESW group) and CXCR4 specific inhibitor AMD3100 control group( AMD group). In vtiro,MSCs were treated with ESW( 500 pulses,10 k V) with the optimal intensity. SDF-1 secretion was detected by ELISA.Osteogenic ability was compared by alizarin red staining and alkaline phosphatase test. In vivo experiment,48 SD rats aged 12 weeks were randomly divided into Ctrl,ESW and AMD groups,with 16 rats in each group. PLGA scaffolds containing these cells were implanted into the right femoral bone defect in each group. AMD group received AMD3100 injection( 1 mg/kg/day) after operation.Samples were taken at the 4 th and 8 th week after operation. Micro-CT,HE staining and immunohistochemistry were used to evaluate bone healing and new bone formation in bone defect area. Results ESW significantly increased the secretion of SDF-1 and osteogenic differentiation of MSCs in vitro and in vivo,and AMD3100 partially inhibited the promotion of ESW. Conclusions ESW can significantly increase the osteogenic differentiation of MSCs. ESW can promote fracture healing through the interaction of secretory protein SDF-1 and CXCR4 receptor,which can promote the formation of new bone in bone defect area. This study provides a theoretical basis for ESW in promoting bone healing in clinical treatment.更多还原