摘要
目的探讨激活态雪旺细胞(activated schwann cells,ASCs)干预骨髓间基质细胞(bone marrow stromal cells,BMSCs)联合异种神经移植体(acellular nerve xenograft,ANX)修复大鼠坐骨神经缺损的效果。方法取大鼠骨髓腔内BMSCs,另取大鼠的ASCs和普通SCs,行原代培养。将两种细胞置于Transwell培养皿中培养并分成三组:BMSCs组,SCs-BMSCs组及ASCs-BMSCs组。在培养皿中培养后第2、4、6和8天,用CCK-8法检测各组BMSCs生物学活性,通过q-PCR检测各组脑源性神经营养因子(BDNF)、神经生长因子(NGF)和碱性成纤维细胞生长因子(bFGF)的mRNA表达水平。在无菌条件下制备10 mm坐骨神经缺损的大鼠模型,制作异种神经支架复合体(兔源胫神经)。将SD大鼠分为三组(n=10):ANX+BMSCs组,ANX+SCs-BMSCs组及ANX+ASCs-BMSCs组。将各组细胞打入支架,复合支架培养3 d,移植到大鼠模型中。术后8周通过神经电生理检测各组患肢感觉神经传导速度(sensory nerve conduction velocity,SCV),使用q-PCR检测神经移植体内神经营养因子(BDNF、NGF和bFGF)表达水平,另比较三组模型的下肢患侧/健侧腓肠肌细胞横截面积比恢复率。结果CCK-8测定发现共培养系统中BMSCs组细胞增殖活性强于单细胞组,第2天和第4天,三组的活性差异无统计学意义(P>0.05),第6天ASCs-BMSCs组增殖活性强于BMSCs组与SCs-BMSCs组,差异有统计学意义(P<0.05),BMSCs组与SCs-BMSCs组之间差异无统计学意义(P>0.05)。通过q-PCR检测BMSCs组中BDNF、NGF及bFGF的mRNA表达最低,ASCs-BMSCs组含量最高(P<0.05)。经8周饲养后的动物实验,通过检测发现,ANX-ASCs-BMSCs组的SCV,ANX内相关神经因子表达量以及ANX-ASCs-BMSCs组的大鼠患侧/健侧腓肠肌肌细胞横截面积均最大。结论ASCs能够促进BMSCs的分化增殖;运用ANX-ASCs-BMSCs促进周围神经功能再生。
Objective To investigate the effect of activated Schwann cells(ASCs)on bone marrow stromal cells(BMSCs)combined with xenograft(ANX)in the repair of sciatic nerve defects in rats.Methods BMSCs were taken from the bone marrow cavity of rats.ASCs and normal SCs were taken for primary culture.The two kinds of cells were cultured in Transwell culture dish and divided into three groups:BMSCs group,SCs-BMSCs group and ASCs-BMSCs group.On the 2nd,4th,6th and 8th day after culture,the biological activity of BMSCs was detected by CCK-8 method,and the mRNA expression of brain derived neurotrophic factor(BDNF),nerve growth factor(NGF)and basic fibroblast growth factor(bFGF)were detected by q-PCR.The rat model of 10 mm sciatic nerve defect was made under aseptic condition,and the heterogenous nerve scaffold complex(tibial nerve from rabbit)was made.SD rats were divided into three groups(n=10):ANX+BMSCs group,ANX+SCs-BMSCs group and ANX+ASCs-BMSCs group.These cells in each group were implanted into the scaffolds and cultured in composite scaffolds for 3 days.The sensory nerve conduction velocity(SCV)was measured by neuroelectrophysiology 8 weeks after operation,and the expression of neurotrophic factors(BDNF,NGF and bFGF)in nerve transplantation was detected by q-PCR.The recovery rate of cross-sectional area ratio of gastrocnemius muscle cells in the affected side/healthy side of the lower limbs was compared among the three groups.Results CCK-8 assay showed that the proliferation activity of BMSCs group was stronger than that of single cell group,and there was no significant difference among the three groups on the 2nd and 4th day(P>0.05).The proliferation activity of ASCs-BMSCs group was stronger than that of BMSCs group and SCs-BMSCs group on the 6th day and the difference was significant(P<0.05).There was no significant difference between BMSCs group and SCs-BMSCs group(P>0.05).The expression of BDNF,NGF and bFGF mRNA was the lowest in BMSCs group and the highest in ASCs-BMSCs group detected by q-PCR(P<0.05).After 8 weeks o
作者
张博闻
赵飞
丁一
黄永禄
巩凡
方芳
段昉
王贺颖
仇莽莽
马宏宁
Zhang Bowen;Zhao Fei;Ding Yi;Huang Yonglu;Gong Fan;Fang Fang;Duan Fang;Wang Heying;Qiu Mangmang;Ma Hongning(Department of Hand and Foot Reconstructive Microsurgery,Ningxia People′s Hospital,Yinchuan 750002,China;Science and Education Office,Ningxia People′s Hospital,Yinchuan 750002,China;Ningxia Medical University,Yinchuan 750004,China)
出处
《中华手外科杂志》
CSCD
北大核心
2020年第5期380-385,共6页
Chinese Journal of Hand Surgery
基金
宁夏自然科学基金(NZ16177、NZ17197)
卫生部手功能重建重点实验室&上海市周围神经显微外科重点实验室开放课题(17DZ2270500)
2018年度医院培育振兴科研项目立项(2018014)
中央高校基本科研业务费专项资金项目(31920190181)。
