化学去细胞异体神经周围复合BMSCs生物蛋白胶复合物促周围神经缺损修复

目的将BMSCs复合在化学去细胞异体神经(chemical extracted acellular nerve allograft,CEANA)周围,观察对CEANA修复周围神经缺损效果的影响。方法成年雄性C57小鼠21只,体重25～30g;成年雄性Balb/c小鼠15只,体重25～30g。取Balb/c小鼠双侧坐骨神经,制备CEANA。取C57小鼠3只,分离培养BMSCs,取5×106个第3代BMSCs添加到500μL生物蛋白胶制备BMSCs生物蛋白胶复合物,共培养3、7、14、21d后,分别取其上清与PC12细胞共培养,观察对PC12细胞的影响。取成年雄性C57小鼠18只,制备小鼠左侧坐骨神经10mm缺损模型,随机分成3组(n=6),分别采用自体神经移植复合生物蛋白胶(A组)、CEANA移植复合BMSCs生物蛋白胶复合物(B组)、CEANA移植复合生物蛋白胶(C组)修复坐骨神经缺损;实验动物右侧切开暴露坐骨神经,作为正常对照。术后行大体观察;术前及术后2、4、6、8周测量小鼠坐骨神经指数(static sciatic index,SSI);术后8周取材计算术侧小腿三头肌湿重恢复率并行小腿三头肌Masson染色观察,吻合口远端神经行甲苯胺蓝染色和透射电镜观察。结果 BMSCs在生物蛋白胶内均匀分布,外观呈球形,培养3d后可见BMSCs呈多个长突起。加入BMSCs生物蛋白胶复合物共培养3、7、14、21d的上清,PC12细胞均分化为类神经元样细胞。术后各组动物切口愈合良好。各组SSI随时间延长逐渐增加,术后4、6、8周A组SSI均高于B、C组,差异有统计学意义(P<0.05);B组略高于C组,但差异无统计学意义(P>0.05)。术后8周,B组小腿三头肌湿重恢复率、有髓神经纤维总数均优于C组,但较A组差,差异有统计学意义(P<0.05);B组小腿三头肌纤维面积、髓鞘厚度均优于C组(P<0.01),而与A组比较差异无统计学意义(P>0.05)。结论在CEANA周围添加BMSCs生物蛋白胶复合物可提高周围神经损伤修复效果。

EFFECT OF CHEMICAL EXTRACTED ACELLULAR NERVE ALLOGRAFT SUPPLEMENTING WITH BONE MARROW MESENCHYMAL STEM CELLS EMBEDDED IN FIBRIN GLUE ON FUNCTIONAL RECOVERY OF TRANSECTED SCIATIC NERVES

Objective To investigate the effect of bone marrow mesenchymal stem cells(BMSCs) embedded in fibrin glue around chemical extracted acellular nerve allograft(CEANA) on the peripheral nerve regeneration.Methods Twenty-one adult male C57 mice(weighing 25-30 g) and 15 adult male Balb/c mice(weighing 25-30 g) were selected.The sciatic nerves were harvested from the Balb/c mice to prepare CEANA.The BMSCs were isolated from 3 C57 mice and were cultured;BMSCs embedded in fibrin glue were cultured for 3,7,14,and 21 ...