人骨形成蛋白4基因修饰的组织工程化骨促进脊柱融合的研究  

Experimental study on spinal fusion induced by hBMP-4 gene modified tissue engineered bone

作　　者：郑召民[1] 董智勇 邝冠明[1] 陈辉[1] 吕游[1] 张奎渤[1] 刘辉[1] 李佛保[1] 

机构地区：[1]中山大学附属第一医院脊柱外科,广州510080 [2]青岛市胶州中心医院骨科

出　　处：《中华外科杂志》2009年第3期197-201,共5页Chinese Journal of Surgery

基　　金：教育部霍英东教育基金“优选资助课题”（94020）;广东省自然科学基金项目（036643）

摘　　要：目的探讨人骨形成蛋白4（hBMP-4）基因修饰的组织工程化骨促进兔脊柱融合的能力，为自体骨寻找理想的移植替代材料。方法重组hBMP-4基因腺相关病毒（AAV-hBMP-4）和重组增强型绿色荧光蛋白（EGFP）基因腺相关病毒（AAV—EGFP）分别按实验确定的最佳感染复数（MOI）值转染兔骨髓间充质干细胞，复合Ⅰ型胶原海绵，构建出表达hBMP-4基因的组织工程化骨及表达对照基因EGFP的人工骨。采用新西兰兔后外侧脊柱融合模型，自身侧侧对照进行实验。14只实验动物随机分为2组，A组右侧为hBMP-4植入侧，左侧为自体骨对照侧。B组右侧为hBMP-4植入侧，左侧为EGFP对照侧。融合术后12周处死动物，评价不同移植物促进脊柱融合的情况。结果术后2组各6只新西兰兔符合标准。术后第12周X线片、三维CT及扪诊检查示hBMP-4侧共11例达到骨性融合（11／12），自体骨侧5例融合（5／6），EGFP侧仅2例融合（2／6）。大体标本观察hBMP-4及自体骨侧新生骨增生明显；EGFP侧成骨量少。结论hBMP-4基因修饰的组织工程化骨在动物体内可以有效地促进成骨，提高脊柱融合率，可望成为一种理想的自体骨移植替代物。Objective To evaluate the efficacy of hBMP-4 gene modified tissue engineered bone graft in the enhancement of rabbit spinal fusion and find an ideal kind of substitute for the autograft bone. Methods Rabbit BMSCs were cultured and transfected with AAV-hBMP-4 using different MOI value. The optimal MOI value were determined by observing cell＇s morphology change. BMSCs were then transfected with AAV-hBMP-4 and AAV-EGFP respectively, following which the transfected cells were evenly suspended in a collagen sponge Ⅰ , and implanted to either side of the L5,6 intertransverse spaces posterolateral in the New Zealand rabbits to induce spinal fusion. Fourteen rabbits were randomly divided into 2 groups. Group 1： AAV-hBMP-4 transfected BMSCs in the right side （hBMP-4 side） and autograft bone in the left side. Group 2：AAV-hBMP-4 transfected BMSCs in the right side（hBMP-4 side） and AAV- EGFP transfected BMSCs in the left side（EGFP side）. Radiographs and three-dimensional CT of the spine, manual palpation, gross and histological examination of the fusion masses for all the animals were performed subsequent to animals having been sacrificed at 12 weeks after surgery. Results Evaluation has been taken in 12 New Zealand rabbits delivered into 2 groups which meet the criterion after operation. Eleven in 12 implemented sides involved hBMP-4 achieved bony fusion, to which 5 in 6 autografted sides was similar. But only 2 in 6 sides in EGFP-group achieved bony fusion meanwhile. Three-dimensional CT scan and palpation also evidenced the results. Bone formation was observed obviously on specimen both in hBMP-4 sides and autografted ones. EGFP-group also got bony integration, but the quantity was small. Conclusion Tissueengineered bone graft constructed from application of hBMP-4 is a fine substitute for autograft. Effective enhancement of bony integration in spinal fusion surgery has been evidenced in vivo.

关 键 词：组织工程 骨形态发生蛋白质类 基因修饰 脊柱融合术 腺相关病毒 

分 类 号：R686[医药卫生—骨科学]