压力调控下骨髓间充质干细胞与脂肪干细胞成软骨能力的比较研究

Comparative study on the modulation effects of mechanical pressure on the chondrogenic differentiation of BMSCs and ADSCs

目的:探讨压力调控对BMSCs和ADSCs两种干细胞在PRF复合生长因子支持下体外成软骨效应的影响。方法:体外分离培养同一个体兔的BMSCs和ADSCs,经鉴定后诱导形成细胞膜片,分别与其同体PRF膜复合制成BMSCs/PRF、ADSCs/PRF双膜复合体。然后分别取其单膜和双膜,并各分为2组(压力刺激组和对照组),各压力刺激组置于细胞加载装置中施加以120 KPa静压力刺激,每天1 h;对照组常规培养。培养后2、4、6 d各取一组,采用实时定量PCR检测增殖基因PCNA mRNA、以及成软骨相关基因Sox-9、Aggrecan、Col-ⅡmRNA的表达水平。结果:与对照组相比,无论是单膜还是双膜复合体,压力刺激均可明显上调BMSCs的细胞增殖基因以及各成软骨相关基因的表达水平(P<0.05);而对ADSCs各基因的表达均无明显作用(P>0.05)。BMSCs各组与ADSCs各组的两两相比,除单膜对照组两者各基因的表达水平无显著差异(P>0.05)外,其余各组的PCNA、Sox-9、Aggrecan及Col-ⅡmRNA表达水平均为BMSCs明显高于ADSCs(P<0.05)。结论:压力对BMSCs的促增殖、促成软骨效应明显强于ADSCs。

AIM： To study the effects of hydrostatic pressure on the chondrogenic differentiation of bone marrow mesenchymal stem cells （BMSCs） and adipose-derived stem cells （ADSCs）. METHODS： BMSCs and AD- SCs were cultured in vitro and platelet-rich fibrin（PRF） membrane was obtained from the same individual rabbit. Cell sheet was induced after identification of the cells BMSCs/PRF and ADSCs/PRF were respectively constructed by com- bining the stem cell sheets and PRF. BMSCs/PRF and ADSCs/PRF in the pressurized groups were stimulated by120 KPa for 1 h per day and those in the control groups were cultured without pressur. The mRNA expression of PCNA, Sox-9, Aggrecan and Col- Ⅱ was analyzed by real-time PCR after 2, 4 and 6 days of culture. RESULTS ： The mRNA expression of proliferation related genes and chondrogenic genes of BMSCs increased significantly in both cell sheets and the compound membranes after hydrostatic pressure loading（ P 〈 0.05 ）, but there was no obvious effect of the pressure on ADSCs（ P 〉 0.05 ）. Aggrecan mRNA and Col-Ⅱ mRNA were significantly higher in BMSCs than in ADSCs（P 〈 0.05） in pressurized culture. CONCLUSION： Mechanical pressure may promote proliferation and chondrogenic dif- ferentiation of BMSCs more significantly than those of ADSCs.