体外震波对激素性股骨头缺血性坏死患者非骨坏死区骨髓间充质干细胞成骨分化的影响  

Extracorporeal shock wave effects on osteogenic differentiation of bone marrow mesenchymal stem cells at the non-necrotic area of patients with steroid-induced avascular necrosis of the femoral head

作　　者：翟磊[1] 孙楠[1] 姜川[1] 徐磊[1] 邢更彦[2] 

机构地区：[1]武警医学院附属医院,天津市300162 [2]武警总医院关节四肢外科,北京市100039

出　　处：《中国组织工程研究与临床康复》2010年第19期3446-3450,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30371430);首都医学发展科研基金资助项目(2002-3037)~~

摘　　要：背景:股骨头缺血性坏死涉及股骨近端骨髓祖细胞脂肪化转变,从而导致骨重建修复困难。能否首先通过体外震波疗法体外诱导自体非骨坏死区的骨髓间充质干细胞为成骨细胞前体,然后移植入自体骨坏死区,从而更加利于坏死区的骨重建?目的:观察体外震波对激素性股骨头缺血性坏死患者非骨坏死区骨髓间充质干细胞分化的影响。方法:取激素性股骨头缺血性坏死患者自愿捐赠骨髓,采用密度梯度离心法结合贴壁法体外分离培养骨髓间充质干细胞。将原代骨髓间充质干细胞无血清培养24h后施以5kV/500频次体外震波干预10min。空白对照组未经体外震波干预。结果与结论:体外震波组细胞传代后进入增殖高峰期更早,呈现出成骨分化趋势;空白对照组细胞存在向其他类型细胞多向分化的表现。体外震波组各时间点(除第1天)细胞增殖速度均明显高于空白对照组(P<0.01)。体外震波组第24天时碱性磷酸酶平均阳性率约90%,空白对照组至第36天才上升至约50%。培养20d茜素红染色示体外震波组矿化结节数为(7.0±1.3)个/视野;空白对照组无明显结节,染色始终为阴性。体外震波组核心结合因子α1mRNA表达在各时间点(除第3天)均明显强于空白对照组(P<0.01);体外震波组OCmRNA表达在各时间点(除第3、6天)均明显强于空白对照组(P<0.01)。结果提示适当强度的体外震波可促进激素性股骨头缺血性坏死患者非骨坏死区骨髓间充质干细胞增殖,并诱导其向成骨细胞方向分化。BACKGROUND：Avascular necrosis of the femoral head involves pimelosis changes of bone marrow progenitor cells on the proximal end of the femur,and leads to difficulties in repairing the bone remodeling.It still remains unclear whether bone marrow mesenchymal stem cells（BMSCs） can be induce-differentiated into osteoblast precursor at autologous non-osteonecrosis region using in vitro extracorporeal shock wave therapy,and then transplanted in autologous osteonecrosis region,contributing to bone remodeling in the zone of necrosis.OBJECTIVE：To investigate the effect of extracorporeal shock wave on the BMSC differentiation in the non-necrotic area of steroid-induced osteonecrosis of the femoral head in patients.METHODS：This study took the bone marrow of steroid-induced osteonecrosis of the femoral head patients who donated voluntarily.BMSCs were isolated and cultured in vitro by density gradient centrifugation and adherence method.Primary cultured BMSCs were cultured with serum-free medium for 24 hours,and intervened by extracorporeal shock wave of 5 kV/500 frequency for 10 minutes.No extracorporeal shock wave was given in the blank control group.RESULTS AND CONCLUSION：Cells after passage in the extracorporeal shock wave group entered the peak phase of proliferation early,showing osteogenic differentiation trend.Cells in the blank control group presented multi-directional differentiation into other cell types.Proliferation speed was significantly greater in the extracorporeal shock wave group than in the blank control group at various time points（except at day 1）（P 0.01）.Mean positive rate of alkaline phosphatase was about 90% at day 24 in the extracorporeal shock wave group,and about 50% at day 36 in the blank control group.At day 20,alizarin red staining exhibited that mineralized nodules in the extracorporeal shock wave group was（7.0 ± 1.3） per field,but no obvious mineralized nodules were found,and staining was always negative in the blank control group.Core binding factor α1 mRNA expressi

关 键 词：体外震波 激素性股骨头缺血性坏死 骨髓间充质干细胞 核心结合因子Α1 骨钙素 

分 类 号：R394.2[医药卫生—医学遗传学]