TGF-β1基因修饰骨髓基质细胞复合Bio-Oss胶原修复犬颌骨缺损的效果

Effect of TGF-β1 Gene Modified Bone Marrow Stromal Cells Combined with Bio-Oss Collagen on Repairing Canine Jaw Defects

目的:观察TGF-β1基因修饰骨髓基质细胞(BMSC)复合Bio-Oss胶原修复Beagle犬颌骨缺损的效果。方法:Beagle犬18条,随机分为3组:种植体+犬BMSC+凝胶+Bio-Oss胶原材料(对照组);种植体+0.5μg/L TGF-β1基因转染犬BMSC+凝胶+Bio-Oss胶原材料(实验组A);种植体+10μg/L TGF-β1基因转染犬BMSC+凝胶+Bio-Oss胶原材料组(实验组B)。术后24周取材,组织观察缺损区恢复情况。RT-PCR检测细胞中OC、BSP及COL-ⅠmRNA的表达情况。Western Blot检测细胞中OC、BSP及COL-Ⅰ蛋白的表达情况。结果:HE染色结果显示实验组A骨连续性及成熟度优于实验组B。RT-PCR结果显示实验组A三种mRNA表达量明显高于对照组,差异具有统计学意义(P<0.05)。实验组B中3种mRNA表达量与对照组相比,不具有统计学意义(P>0.05)。此外,Western Blot结果中实验组A的蛋白表达量与对照组和实验组B相比明显增加,具有统计学意义(P<0.05),实验组B较对照组略有增加,但差异不具有统计学意义(P>0.05)。结论:低浓度的TGF-β1基因可较好地促进BMSC向成骨分化。

Objective:To observe the effect of TGF-β1 gene modified Bone Marrow Stromal Cells(BMSC)combined with Bio-Oss collagen on jaw defects reconstruction.Methods:18 beagle dogs were randomly divided into three groups.Control group:implant+BMSC+gelatin+Bio-Oss collagen group A:implant+BMSC transfected with 0.5μg/L TGF-β1+gelatin+Bio-Oss collagen;group B:implant+BMSC transfected with 10μg/L TGF-β1+gelatin+Bio-Oss collagen.Histological view was applied to measure the repair effect.Meanwhile,the mRNA and protein expressions of OC,BSP and COL-I were detected through RT-PCR and Western blot,respectively.Results:HE staining showed that bone continuity and maturity in group A were superior to those in group B.RT-PCR results showed that the expressions of mRNA in the experimental group A were significantly higher than those in the control group(P<0.05).However,compared with the control group,the mRNA expression levels in the experimental group B were not statistically significant(P>0.05).In addition,Western blot results showed that expressions of OC,BSP,and COL-I protein in the group A were significantly higher than those of control group and group B(P<0.05),while that of group B was slightly higher than that of control group,but without statistically significant(P>0.05).Conclusion:0.5μg/L TGF-β1 gene has a better repair effect on promoting osteogenic differentiation of BMSC.