目的使用基因枪进行含有BMP-2基因真核表达质粒的局部基因转移以治疗新鲜骨缺损。方法选健康3月龄新西兰大白兔72只,建立新鲜兔桡骨中段骨缺损模型,按缺损长度分组:(1)1.5cm组;(2)2.0cm组;(3)2.5cm组,各24只。各组按干预又分为BMP-2基因转染组(治疗组)和自然愈合组(对照组),各12只。新鲜骨缺损于截骨后立即使用基因枪进行BMP-2基因局部转染,对照组截骨后自然愈合。于转染后1、3、4、9w拍摄X线片,1、3、4w取骨折间组织行BMP-2的Western blot检测,于1、4和9w时取大体标本肉眼观察,评价愈合情况。结果 (1)大体标本观察,治疗组骨痂形成数量普遍多于对照组。(2)治疗组与对照组转染后1、3、4、9w的Lane-Sandhu X线评分,1w时差异无统计学意义(P〉0.05)。3、4、9w时差异有统计学意义(P〈0.05):治疗组优于对照组。(3)BMP-2浓度定量,除1.5cm组1w时治疗组与对照组差异无统计学意义。其余各时间点治疗组与对照组BMP-2浓度差异均有统计学意义(P〈0.05):治疗组优于对照组。结论应用基因枪介导BMP-2基因局部转移治疗新鲜骨缺损效果明确。
Abstract: Objective To explore the therapy of fresh bone defect by using gene gun to transfer the local gene containing BMP-2 gene eukaryotic expression plasmid. Methods Select 72 healthy three-month old New Zealand white rabbits; set up a new model of rabbit bone defect in middle of radial; group rabbits ac- cording to the defect length: (1) 24 rabbits in 1.5 cm group, (2) 24 rabbits in 2.0 cm group, (3) 24 rab- bits in 2.5 cm group. Each group includes two subgroups according to the intervention: 12 rabbits in treat-ment group (BMP-2 gene transfection group) and 12 rabbits in control group. Through the application of gene gun for BMP-2 local transfection after bone cutting immediately, the control group with natural heal- ing after bone cutting. X-ray examinations are taken at 1 week, 3 weeks, 4 weeks and 9 weeks after trans-fection, and the tissues between the bone defect are taken to do BMP-2 Western blot detection at 1 week,3 weeks and 4 weeks; and radius specimens are weeks, conduct evaluation of the healing. Results taken Ior gross observation at 1 week,4 weeks and 9 weeks, conduct evaluation of the healing. Results (1) Gross specimen observation: bone callus Iormauon of treatment group is generally more than the control group. (2) Lane-Sandhu X-ray score: there is no sta-tistically significant difference at 1 w (P 〉0.05) ; However, the difference is statistically significant at 3, 4, 9 w between treatment group and control group (P 〈0.05) :treatment group is superior. (3) BMP-2 concentration quantitatively: there was no statistically significant difference at 1 w in the 1.5 cm group; and the rest of each time point, BMP-2 concentration differences had statistical significance (P〈0.05) : treatment group was superior. Conclusion The application of gene gun mediated BMP-2 gene to do local gene transfer in therapy of fresh bone defect is efficacious.