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  被引量:5

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

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作  者:赵喆[1] 王玉[1] 彭江[1] 赵斌[1] 赵庆[1] 刘炎[1] 任志午[1] 詹胜峰[1] 张莉[1] 许文静[1] 卢世璧[1] 

机构地区:[1]解放军总医院全军骨科研究所,北京100853

出  处:《中国修复重建外科杂志》2011年第4期488-493,共6页Chinese Journal of Reparative and Reconstructive Surgery

基  金:国家高技术研究发展计划(863)资助项目(2009AA03Z312);国家自然科学基金资助项目(30571875);全军医药卫生科研基金资助项目(06Z057);国家科技支撑计划资助项目(2009BAI87B02)~~

摘  要: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。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 days.Then the supernatant was obtained and co-cultured with PC12 cells for 2 days to observe the PC12 cell growth in vitro.The other 18 C57 mice were used to establish the left sciatic nerve defect models of 10 mm and divided into 3 groups:autogenous nerve graft with fibrin glue(group A,n=6),CEANA graft with BMSCs(5 × 106) embedded in fibrin glue(group B,n=6),and CEANA graft with fibrin glue(group C,n=6).The right sciatic nerves were exposed as the controls.At 2,4,6,and 8 weeks,the mouse static sciatic index(SSI) was measured.The histomorphometric assessment of triceps surae muscles and nerve grafts were evaluated by Masson staining,toluidine blue staining,and transmission electron microscope(TEM) observation at 8 weeks after operation.Results BMSCs were uniform distribution in fibrin glue,which were spherical in shape,and the cells began to grow apophysis at 3 days.PC12 cells differentiated into neuron-like cells after addition supernatant co-cultured after 2 days.Incisions healed well in each group.At 2,4,6,and 8 weeks,the SSI increased gradually in 3 groups.SSI in group A was higher than that in groups B and C at 4,6,and 8 weeks after operation(P 0.05).SSI in group B was slightly higher than that in group C,but had no significant difference(P 0.05).At 8 weeks,the wet weight recovery rate of triceps surae muscles and fibers number of myelinated nerve were better in group B than in group C,but worse in group B than in group A,showing significant differences(

关 键 词:化学去细胞异体神经 BMSCS 神经营养因子 神经修复 大鼠 

分 类 号:R651.3[医药卫生—外科学]

 

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