中文摘要：

目的：观察转人胰岛素样生长因子Ⅰ基因同种异体组织工程软骨对兔膝关节软骨全层缺损的修复作用。 方法：实验于2004～08／2005—08在军事兽医研究所病毒室及长春生物制品研究所实验室完成。①取出生28d新西兰白兔的关节软骨，采用机械分离及Ⅱ型胶原酶消化法获得分离的兔软骨细胞，用单层培养扩增。②用脂质体法使胰岛素样生长因子Ⅰ基因转染兔关节软骨细胞。③以经过转染的兔关节软骨细胞作为种子细胞，牛Ⅰ型胶原作为支架，体外构建转基因同种异体组织工程软骨。④5个月龄的新西兰白兔80只制备关节软骨缺损模型，分别移植入不同组别的移植物：空白对照组、单纯支架组、单纯软骨细胞附和支架组、经过转染的软骨细胞附和支架组、自体软骨移植组，分别于术后4，8，16，24周处死各组动物4只取材，观察兔膝关节软骨全层缺损的修复情况。结果：①G418培养液筛选结果：通过G-418对于兔关节软骨细胞的最小致死剂量的测定得出，G418对软骨细胞的最小致死剂量为0．4g／L。转染后软骨细胞经0．4g／L G418筛选，2周后得到抗G418的阳性细胞克隆，而未转染细胞在含G418的培养液中逐渐全部死亡，初步证明人胰岛素样生长因子Ⅰ基因的转染成功。②原位杂交检测结果：转染软骨细胞胞浆中有氯化镍紫蓝色颗粒阳性杂交信号说明有人胰岛素样生长因子1mRNA的表达；未转染细胞则未见阳性信号。③免疫组织化学检测结果：在转染的软骨细胞胞浆中有棕黄色颗粒样阳性信号，说明有Ⅱ型胶原蛋白表达，证明稳定表达人胰岛素样生长因子Ⅰ的转染软骨细胞保持Ⅱ型胶原的表达。④软骨缺损修复后的组织学评价结果：在试验所观察的24周内，软骨基本稳定，而且转胰岛素样生长因子Ⅰ基因组织工程软骨显示出强烈的软骨基质分泌能力。?

英文摘要：

AIM： To investigate the effect of allogenic tissue-engineered cartilage transfected by human insulin-like growth factor 1 （IGF-1） on the repair of full-thickness defects of articular cartilage of knee of rabbits. METHODS： This experiment was conducted in the Laboratory of Research Institute of Military Hippiater and Changchun Institute of Biological Products from August 2004 to August 2005.①The cartilages of New Zealand rabbits born in 28 days were taken, and the mechanical isolation method and Ⅱ collagenase digestion were used to get the isolated chondrocyte. The number of chondrocyte was enlarged with monolayer cultivation. ② The chondrocyte were transfected with liposome encapsulated human IGF-1 gene. ③The tissue-engineered cartilage with the transfected chondrocyte as the seed cells and the cattle collagen Ⅰ as the scaffold of was constructed. ④ Eighty New Zealand white rabbits aged 5 months were selected to establish the models of cartilage defects, which was implanted different groups of graft： Blank control group, scaffold group, chondrocyte adherence scaffold group, transfected chondrocyte adherence scaffold group and autoallergic cartilage transplanted group. Four rabbits in each group were sampled after killed at 4, 8, 16 and 24 weeks after operation to observe the repair condition of full-thickness defects of knee. RESULTS： ①G418 screening： The smallest causing death dose of G418 for chondrocyte was 0.4 g/L. The chondrocyte were screened by 0.4 g/L G418 culture solution, and two weeks later, the transfected chondrocyte formed anti-G418 positive cell clone and the untransfected ones died,primarily indicating the transfection of IGF-1 was successful. ② Hybridization in situ： Amethyst niekel chloride particle was found in the kytoplasm of infected chondrocyte, showing there were mRNA expressions of human IGF-1 gene; nothing was found in the normal chondrocyte. ③Immunohistochemical detection： Buffy particle was found in the kytoplasm of transfected chondrocyte, indic