目的:通过观察静压力刺激对骨髓间充质干细胞(BMSCs)体外增殖及成软骨能力的影响。方法:全骨髓贴壁分离法分离大鼠BMSCs,经表面标志物检测和成脂、成骨诱导分化鉴定后,随机分为0(对照组)、90、120、150kPa4个大组;每一大组按每天加载持续时间再分为1h/d和6h/d2个亚组。然后在细胞液压加载装置内分别给予相应的静压力刺激,分别于实验后2、4d时用RT—PCR检测BMSCs成软骨相关基因SOX-9、Aggrecan、Col-Ⅱ以及细胞增殖基因PCNAmRNA表达水平。结果:相同加压条件下不同时间比较,连续压力刺激2d时BMSCs增殖基因及成软骨基因的表达量均明显高于4d组(P〈0.05)。同样在连续加压2d时间点内,每天加压1h组BMSCs的增殖基因及成软骨基因的表达量均明显高于6h组(P〈0.05),其中,每天压力刺激1h连续2d的条件下90kPa压力组的PCNA、SOX-9、Aggrecan、Col一ⅡmRNA的表达均显著高于相同刺激时间的120kPa和150kPa压力刺激组(P〈0.05)。结论:在诸多压力刺激条件中,90kPa静压力每天1h连续作用2d促BMSCs增殖与成软骨向分化作用最显著。
AIM. To investigate the effects of hydrostatic pressure on the proliferation and chondrogenic dif- ferentiation of bone marrow mesenchymal stem cells (BMSCs). METHODS : BMSCs were obtained from SD rats by the whole bone marrow adherent separation culture method and identified by cell surface marker detection, adipogenic and osteogenic induction assays. Then the BMSCs were treated by hydrostatic pressure at 0, 90 , 120 and 150 kPa for lh/d and 6h/d, for 2 and 4 d respectively in a hydraulic pressure - controlling cellular-strain unit. Gene expressions of PCNA,SOX-9,Aggrecan and COL-U were analyzed by real-time PCR. RESULTS: Gene expressions of PCNA, SOX-9, Aggrecan and COL- II in the 2 consecutive-day-pressure - treatmemn ceils were higher than those in the 4 day treatmemn cells (P 〈0.05). Within the 2 day treatmemn groups, lh/d treatmemn of pressure at 90, 120 or 150 kPa showed stronger effects than 6 h/d ( P 〈 0.05). 2 consecutive-day-pressure-treatmemn at 90 kPa for lh/d showed the highest promotion effects on the expressions of the genes in BMSCs (P 〈 0.05). CONCLUSION: Adequate hydrostat- ic pressure may promote the proliferation and chondrogenic differentiation.