应用转染BMP7基因组织工程软骨修复兔膝关节软骨缺损观察

Restore cartilage deficiency of rabbit knuckle with Bone morphogenetic protein 7 gene-transfected cartilage

目的构建转染骨形态发生蛋白-7（BMP7）基因组织工程软骨，观察对家兔膝关节软骨缺损的修复作用。方法将转染BMP7基因的软骨细胞（5×10^9个／L）接种于胶原-纤维蛋白凝胶的支架中，培养14d后移植于家兔膝关节软骨表面，直径为5．0mm，深达软骨下骨板的全层软骨缺损部位。移植后4、8、12周处死家兔，通过创面形态观察、组织形态的光镜观察、BMP7基因在移植部位的表达，评价转染BMP7基因组织工程软骨对兔膝关节软骨缺损的修复作用。结果转染BMP7和未转染BMP7基因组织工程软骨培养物的体积分别为9min×9mm×2mm和8mm×8mm×2mm。转染BMP7基因组移植于家兔膝关节软骨后4、8、12周，均有软骨缺损修复作用，且移植部位有红褐色BMP7mRNA和棕红色BMP7蛋白表达。移植后12周修复效果佳，优于未转染BMP7基因组。结论转染BMP7和未转染BMP7基因组织工程软骨对兔膝关节软骨缺损均有修复作用，但转染BMP7基因组织工程软骨修复作用更强。

Objective To observe the restoration of Bone morphogenetic protein 7（BMP7） gene-transfected cartilage on cartilage deficiency of rabbit knuckle. Methods BMP7 gene-transfected chondrocytes were cultivated in collagen gelatin bracket（5 ×10^9/L）. Rabbits were subjected to cartilage deficiency of knuckle. The deficiency, in a diameter of 5.0 mm, was whole cartilage plate deep. BMP7 gene-transfected chondrocytes were transplanted into defective section after 14 days culture. The rabbits were executed respectively 4, 8 and 12 weeks after transplantation. Then restoration of BMP7 gene-transfected cartilage on cartilage deficiency of rabbit knuckle was evaluated with different methods. The morphological changes of the surface of the wound, histomorphologic changes were observed under optical microscope, and also the expression of BMP7 gene was measured at transplant section. Results The volumes of cartilage cultures of BMP7 gene-transfected and non-BMP7 gene-transfected were 9 mm × 9 mm × 2 mm and 8 mm × 8 mm × 2 mm, respectively. BMP7 gene-transfected chondrocytes repaired cartilage deficiency either 4 or 8 and 12 weeks after transplanted. BMP7 mRNA expressed at the transplanted section. The restoration effect of BMP7 group was excellent 12 weeks after transplantation and significantly superior to that of non-BMP7 group. Conclusions BMP7 gene-transfected cartilage and non-BMP7 cartilage have restoration effects on cartilage deficiency of rabbit knuckle. BMP7 gene-transfected cartilage is superior to that untransfected cartilage.