Leng Ping, Ding Chang-rong, Zhang Hai-ning, Wang Ying-zhen
Department of Pharmacy, the Affiliated Hospital, School of Medicine, Qingdao University, Qingdao 266003, China.
Knee. 2012 Dec;19(6):804-11. doi: 10.1016/j.knee.2012.03.009. Epub 2012 Apr 6.
To investigate a compound technique including gene therapy, injectable tissue engineering and Mosaicplasty to reconstruct large osteochondral defect.
Plasmid vector containing hIGF-1 cDNA was created and transfected into BMSCs in vitro with FuGene6. After gene expression determination, cells were mixed with calcium alginate gel. Osteochondral defects were created on the femoral condyle of goats in a diameter of 6mm. Osteochondral plugs were harvested from the intertrochlea groove and pressed into the recipient sites in a mosaic mode. Gene modified BMSCs-scaffold complex was applied to fill the residual defects. Control groups were also set up. At 4 and 16 weeks, specimens were investigated in gross and under microscopy, electromicroscopy and MRI detection.
hIGF-I gene was expressed effectively with the peak concentration at 34.75 ng/ml. Subchondral bone and cartilage were integrated well in gene enhanced Mosaicplasty group. The reconstructed tissue filled up the gaps between columns, which appeared better than other groups. The regenerated cartilage was integrated with neighbor tightly in regular arrange. Extracellular matrix distributed evenly and deeply stained by alcian blue. Quantitative histologic assessments showed higher score in gene enhanced Mosaicplasty group. Glycosaminoglycan assay revealed no difference between groups involving Mosaicplasty. MRI analysis demonstrated the healing process between the subchondral bone other than control groups.
hIGF-I gene enhanced tissue engineering can modify the outcome of Mosaicplasty to reconstruct large osteochondral defects in weight-bearing region.
研究一种包括基因治疗、可注射组织工程和镶嵌植骨术在内的复合技术，用于重建大的骨软骨缺损。
构建含有人胰岛素样生长因子-1（hIGF-1）cDNA的质粒载体，并用FuGene6在体外将其转染到骨髓间充质干细胞（BMSCs）中。在确定基因表达后，将细胞与海藻酸钙凝胶混合。在山羊股骨髁上制作直径为6mm的骨软骨缺损。从髁间沟获取骨软骨栓，以镶嵌模式压入受体部位。应用基因修饰的BMSCs-支架复合物填充剩余缺损。同时设立对照组。在4周和16周时，对标本进行大体观察、显微镜、电子显微镜和磁共振成像（MRI）检测。
hIGF-1基因有效表达，峰值浓度为34.75ng/ml。基因增强镶嵌植骨术组的软骨下骨和软骨整合良好。重建组织填充了柱状结构之间的间隙，表现优于其他组。再生软骨与相邻组织紧密整合，排列规则。细胞外基质分布均匀，阿尔辛蓝染色深。定量组织学评估显示基因增强镶嵌植骨术组得分更高。糖胺聚糖检测显示涉及镶嵌植骨术的各组之间无差异。MRI分析显示除对照组外软骨下骨的愈合过程。
hIGF-1基因增强组织工程可改善镶嵌植骨术的效果，用于重建负重区域的大骨软骨缺损。
