生长转化因子-β3转染骨髓基质干细胞联合脱钙骨基质修复软骨缺损

Experimental study on repair of cartilage defect with BMSC transfected with TGF-β3 in combination with DBM

目的通过生长转化因子-β3(TGF-β3)转染骨髓基质干细胞(BMSC)后,以脱钙骨基质(DBM)为载体共同修复软骨缺损。方法首先在将BMSC和DBM在体外混合培养21 d,实验组中加入TGF-β3转染后的p3 BMSC,对照组中加入未转染的p3 BMSC,7、14、21 d取出DBM块,通过逆转录-聚合酶链反应(RT-PCR)检测COL(Ⅱ)、COL(X)和Aggrecan的量,第21天取出的DBM晾干后称重,计算净增长重量。然后将经过体外混合培养7 d的DBM植入兔的软骨缺损模型中,实验组在培养的过程中加入转染后的BMSC,对照组加入未经转染的BMSC。3、6、9周处死后取材,进行肉眼和组织学观察。结果在体外的混合培养的实验中,TGF-β3转染后的BMSC能显著的增加DBM上的软骨生成量,增加量约为对照组的6倍。同时还大大增加了正常软骨基质COL(Ⅱ),COL(X)和Aggrecan的产生。在体内实验中,加入TGF-β3转染后BMSC的实验组的软骨修复效果明显好于未加入TGF-β3的对照组。结论TGF-β3转染BMSC后,再联合DBM,有更强和迅速的软骨修复能力。

Objective To repair the huge cartilage defect with BMSC transfected with TGF-β3 in combination with DBM as a cartier. Methods DBM and BMSC were co-cultured in vitro for 21 days. In experiment group DBM and p3 BMSC transferred with TGF-β3 were co-cultured in incomplete ehondrogenie media, and in control group DBM and p3 BMSC without transfection were cultured in the same media. In the composite of DBM/BMSC at 7th, 14th and 21 st day, the mRNA expression of COL（ U ）, COL（X） and Aggrecan was detected by RT-PCR. The composites harvested at 21st day were weighed and the increased net weight was calculated. In the in vivo study, the composites of DBM/BMSC co-cultured in vitro for 7 days were implanted into the cartilage defect of rabbits. In the in vivo experiment group the composites were cultured in the presence of BMSC transferred with TGF-β3, and in the in vivo control group, there was no tansfection with TGF-β3. Specimens were collected at 3rd, 6th ,9th week, and a series of general and histomorphologieal observations were performed. Results In vitro study of eo-eulutre, BMSC transfected with TGF-β3 could signifieandy increase the ehondrogenesis on the surface of the composites by about 6- fold as compared with control group, and the expression of COL（ Ⅱ ）, COL（X） and Aggrecan. In vivo, the effect of cartilage defect repari in the experiment group was better than in the control group. Conclusion The repair of cartilage with DBM/BMSC transfected with TGF-β3 can be improved obviously.