含BMSCs的ANX移植联合G-CSF修复大鼠坐骨神经缺损的实验研究

Experimental sciatic nerve repair by ANX combined with BMSCs and G-CSF intervention in rats

目的:探讨复合骨髓基质细胞(BMSCs)的脱细胞异种神经移植体(ANX)移植及联合应用粒细胞集落刺激因子(G-CSF)修复大鼠坐骨神经缺损的效果。方法:将大鼠BMSCs种植到ANX内复合培养,制备大鼠坐骨神经10 mm缺损模型,SD大鼠随机分为4组(n=10):ANX移植组、ANX移植联合应用G-CSF组、复合BMSCs的ANX移植组和复合BMSCs的ANX移植联合应用G-CSF组。术后8周,行坐骨神经功能指数(SFI)和电生理检测;应用免疫荧光染色和实时荧光定量PCR检测再生神经内神经丝和神经营养因子的表达。结果:含BMSCs的ANX移植联合应用G-CSF组SFI、神经传导速度、再生纤维相对密度增高(P<0.05),神经营养因子表达增加(P<0.05)。结论:含BMSCs的ANX移植联合应用G-CSF可显著促进周围神经再生。

Objective： To discuss the effectiveness of the acellular nerve xenograft（ANX） implanted with bone marrow stromal cells（ BMSCs） and combined with granulocyte colony stimulating factor（ G-CSF） intervention on repair of sciatic nerve defect in rats. Methods： BMSCs of rats were injected into ANX to culture. The right sciatic nerves of Sprague-Dawley rats were excised to form 10 mm defects,then the rats were randomly divided into four groups（ n = 10）： ANX group,ANX combined with G-CSF group,ANX implanted with BMSCs group,and ANX combined with BMSCs and GCSF intervention group. At 8 weeks post-transplantation,sciatic nerve function index（ SFI） and electrophysiological indexs were detected. To test the expression of neurofilament-200（ NF-200） and neurotrophic factors in regenerative sciatic nerves by immunofluorescence and real time PCR. Results： SFI,nerve conduction velocity and the relative density of regenerated nerve fibers in the ANX combined with BMSCs and G-CSF intervention group were increased（ P 0. 05）,the expression of protein and m RNA levels of NGF and BDNF were increased significantly（ P 0. 05）. Conclusion： The ANX implanted with BMSCs and combined with G-CSF invention promoted the regeneration of peripheral nerves obviously.