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低强度脉冲超声波对大鼠脂肪干细胞增殖及成骨分化的影响 被引量：3

The effects of low intensity pulsed ultrasound on the proliferation and osteogenic differentiation of rat adiposetissue-derived stem cells
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摘要 目的探讨低强度脉冲超声波（LIPUS）对体外分离培养的大鼠脂肪源干细胞（ADSCs）增殖及成骨分化的影响。方法取4周龄Sprague－Dawley大鼠腹股沟处脂肪组织，分离出脂肪干细胞并体外培养，经免疫磁珠分选、流式细胞仪鉴定ADSCs表面标志后，取第3代ADSCs，经LIPUS作用不同时间后，采用流式细胞仪检测细胞周期，计算细胞增殖指数，采用CCK-8法比较细胞增殖活性。取第3代ADSCs，设置阴性对照组、LIPUS组、成骨诱导组及LIPUS联合成骨诱导组，于LIPUS作用7d、14d时测定各组细胞碱性磷酸酶（ALP）活性；于LIPUS作用21d时采用Von kossa染色比较各组细胞成骨分化能力。结果第3代ADSCs高表达CD29、CD44，低表达CD34，符合ADSCs表面标志物特征；LIPUS组细胞增殖指数较对照组明显提高，细胞增殖加快。与阴性对照组比较，LIPUS组于实验进行7d、14d后ALP活性明显升高；LIPUS作用能促进ADSCs致矿化结节形成，但其矿化结节数量不及LIPUS联合成骨诱导组。结论LIPUS能加速ADSCs增殖，促进ADSCs向成骨细胞分化。 Objective To investigate the biological effects of low intensity pulsed ultrasound （LIPUS） on the proliferation and osteogenic differentiation of adipose tissue-derived stem cells （ADSCs） in vitro. Methods Primary ADSCs were harvested from the inguinal fat pads of 4-week-old female Sprague-Dawley rats, cultured in vitro and purified by magnetic-activated cell sorting. Surface ADSC markers were identified by flow cytometry. LIPUS at 100 mW/cm2 was used to stimulate the cultured cells. Flow cytometry was performed for cell cycle analysis. Cellular proliferation was evaluated via CCK8 chromatometry, and a proliferation index was calculated. ADSCs were assigned to 4 groups： a negative control group, a LIPUS group, an osteoiuduction group and a LIPUS plus osteoinduction group, and treated accordingly. Alkaline phosphatase （ALP） activity was determined at the 7th and 14th day in each group, and calcium nodes were marked by Von Kossa staining. The levels of osteogenic differentiation in the different groups were evaluated. Results The ADSCs of passage 3 expressed CD 34^low, and CD29^high CD44^high , which was consistent with the characteristics of ADSC surface markers. Proliferation was upregulated significantly in the LIPUS group compared with the negative control group. ALP activity was also elevated significantly and it resulted in mineralization. The highest mineralization rate was observed in the LIPUS plus osteoinduction group. Conclusions LIPUS not only can stimulate the proliferation of rat ADSCs, it also promotes their osteogenic differentiation.
作者 江渟 郭风劲 许涛 杨克勤
机构地区 华中科技大学同济医学院附属同济医院骨科 华中科技大学同济医学院附属同济医院康复医学科 安徽医科大学第二附属医院骨科
出处 《中华物理医学与康复杂志》 CAS CSCD 北大核心 2011年第6期408-412,共5页 Chinese Journal of Physical Medicine and Rehabilitation
关键词 低强度脉冲超声波 大鼠 脂肪源干细胞 增殖 成骨分化 Low intensity pulsed ultrasound Rats Adipose tissue-derived stem cells Proliferation Osteogenic differentiation
分类号 R329.2 [医药卫生—人体解剖和组织胚胎学]
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参考文献28

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共引文献22

同被引文献82

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引证文献3

二级引证文献27

中华物理医学与康复杂志
2011年 第6期

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