腺病毒介导的人骨形态发生蛋白2基因修复兔桡骨缺损

Repair of radial bone defects of rabbits by adenovirus mediated human bone morphogenetic protein-2 gene

目的 评价腺病毒介导的人骨形态发生蛋白 2 (Adv -hBMP - 2 )基因对兔骨髓间充质干细胞 (BMSCs)的诱导成骨活性及修复长骨干骨缺损的效果。 方法 ( 1)抽取兔骨髓行BMSCs培养 ,经Adv-hBMP - 2转染后分别行免疫沉淀加Western印迹法和碱性磷酸酶 (ALP)检测及vonKossa染色 ,并行裸鼠肌内诱导成骨试验。 ( 2 )修复兔桡骨缺损 :15只兔 3 0侧、1.5cm的骨缺损分为Adv -hBMP - 2转染BMSCS加珊瑚及胶原载体组、β半乳糖苷酶 (Adv - βgal)转染BMSCs加载体组、未转染BMSCs加载体组、载体组和未治疗组 ,术后行X线、组织学检查。 结果 ( 1)Adv -hBMP -2组BMSCs表达hBMP - 2 ,其ALP活性升高 ,并有钙结节形成 ,裸鼠肌内注射后有异位成骨。 ( 2 )在兔桡骨缺损处 ,Adv -hBMP - 2转染组有明显骨痂形成 ,5个组的愈合率分别为 4/5、2 /5、2 /5、0 /5、0 /5。 结论 Adv -hBMP -

Objective To evaluate effectiveness of adenovirus mediated human bone morphogenetic protein-2 gene (Adv-hBMP-2) in repair of segmental bone defect by being transferred to bone marrow derived mesenchymal stem cells (BMSCs) of rabbits. Methods The rabbit BMSCs from bone marrow were collected and cultured. After transduced by Adv-hBMP-2, these BMSCs were evaluated by immunoprecipitation plus Western blotting, ALP activity assay and von Kossa staining, and injected into muscles of nude mice to test their osteoinductivity. Thirty radial bone defects (1.5 cm) from 15 rabbits were divided into 5 groups (6 sides per group): Adv-hBMP-2 transduced BMSCs-collagen-coral bone substitute group (Group I), Adv-βgal transduced BMSCs-collagen-coral group (Group II), untransduced BMSCs-collagen-coral group (Group III), collagen-coral group (Group IV) and untreated group (Group V). Roentgenographic and histological examinations were performed at various time after implantation. Results After hBMP-2 gene transduction, the BMSCs expressed and secreted hBMP-2 with increased activity of ALP and formation of calcified nudes. BMSCs transduced by hBMP-2 induced ectopic bones in the muscles of nude mice. In GroupⅠ, the radial bone defects of rabbits were filled with much more bony callus. The healing rates of Groups Ⅰ, Ⅱ, Ⅲ, Ⅳ and V were 4/5, 2/5, 2/5, 0/5 and 0/5, respectively. Conlusions Adv-hBMP-2 gene transferred to BMSCs is an excellent method for clinical treatment of bone defects.