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作 者:王五洲[1] 邢更彦[1] 井茹芳[2] 匡正达[1] 姜川[1] 周安全[3] 杨增前
机构地区:[1]武警总医院骨科,北京市100039 [2]武警总医院超声科,北京市100039 [3]武警8662部队卫生队,新疆新源县835800
出 处:《中国康复理论与实践》2006年第5期372-375,共4页Chinese Journal of Rehabilitation Theory and Practice
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(20023037)。
摘 要:目的确定冲击波(ESW)对成人骨髓间充质干细胞(hMSCs)增殖分化的影响及体外干预的最佳能量等级。方法采用密度梯度离心法提取hMSCs;体外培养成功后应用四甲基偶氮唑盐法(MTT)确定ESW干预的合适剂量;利用倒置相差显微镜观察干预后hMSCs形态、贴壁率和倍增时间等变化,并应用酶细胞化学法检测碱性磷酸酶(ALP)含量的变化。结果5kV(100次)的体外冲击波可以明显促进hMSCs的生长(P<0.01)。干预后的hMSCs贴壁率升高,倍增时间缩短,增殖分裂加快,ALP总体水平高于对照组(P<0.05)。结论ESW干预能够提高hMSCs的增殖及成骨活性;5kV(100次)左右是其最佳能量。Objective To explore the mechanism of extracorporeal shock wave (ESW) in treating osteogenic disorders and the ideal energy level. Methods After success in marrow aspiration from patients' iliac crest, hMSCs were isolated by Pereoll density gradient centrifugation and cultured in Dulbecco's modified Eagle's medium in a 5% CO2 and 37℃ incubator. Optimal ESW dose was determined by MTT of kinase-marked cytobiology. After hMSCs were exposed to ESW, their morphocytologic change, rate of adherence and doubling time were observed with IPCM. Enzyme cytochemistry reaction for the activity of alkaline phosphatase was also examined. Results ESW of 5 kV and 100 times could increase cells' viability and proliferation (P 〈 0.01), but higher than 7 kV would inhibit them. Rate of adherence of hMSCs in exposure group of passage 5 reached to 61.54%, which was significantly different from control group (P 〈 0.05). Compared with control group, the MSCs' doubling time was short for 1.72 d (P 〈 0.05). The curve of normal alkaline phosphatase activity of hMSCs was llke type S, but ESW shortened its latent period, and promoted its peak time, which was significantly different from control group. Conclusion ESW of 5 kV and 100 times can optimally promote the proliferation and activity of osteogen of hMSCs in vitro.
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