胶原神经导管对外周神经缺损修复的实验研究  

Experimental study on collagen nerve conduit for repairing peripheral nerve defects

作　　者：赵红斌[1] 马敬[1] 杨银书 刘旭东[1] 董菊子[1] 马慧[1] 

机构地区：[1]兰州军区兰州总医院,甘肃兰州730050 [2]兰州军区疾病控制中心,甘肃兰州730021

出　　处：《生物医学工程与临床》2010年第2期100-104,F0002,共6页Biomedical Engineering and Clinical Medicine

基　　金：兰州军区医药卫生科研项目(LXH-2007007)

摘　　要：目的探讨组织工程化胶原神经导管的构建及对外周神经缺损的修复。方法分别取10g胶原蛋白利用自制模具分别制备含0.25g川芎嗪和无川芎嗪的胶原神经导管(2%京尼平交联);对交联前后的胶原神经导管进行拉伸实验并评价其力学特性;用扫描电子显微镜观察胶原神经导管交联前后的空间结构;将大鼠骨髓间充质干细胞(MSC)与细胞外基质(ECM)混合后接种于导管内腔,用扫描电子显微镜观察细胞与材料的复合情况;质量浓度10g/L的川芎嗪诱导MSC7d后,荧光免疫化学方法鉴定MSC分化为神经元细胞;用8只Wister大鼠复制坐骨神经10mm缺损模型,复合MSC的神经导管连接缺损神经,其中4只为无中药导管作为对照,90d后处死大鼠观察再生神经形态,免疫组织化学鉴定其功能。结果京尼平交联前后胶原纤维结构发生明显改变,交联后胶原纤维排列致密,胶原纤维之间形成不规则孔隙且韧性增强;力学实验结果表明:交联前后的胶原神经导管的最大载荷和断裂载荷分别为(0.23±0.09)、(0.20±0.12)N和(0.76±0.15)、(0.69±0.17)N,两者差异具有显著统计学意义(P﹤0.01);川芎嗪能促进MSC表达神经元特异性烯醇酶并向神经细胞分化;神经导管与MSC复合培养两者具有良好的组织相容性;复合川芎嗪的胶原神经导管能促进缺损神经的修复。结论构建的组织工程化胶原神经导管能有效修复外周神经缺损。Objective To investigate the construction of tissue engineering collagen nerve conduits and repair peripheral nerve defects.Methods The collagen protein was separated and purified from bovine achilles tendon tissue with salting-out technique,collagen nerve conduits contained 0.25 g ligustrazine and non-ligustrazine were fabricated from 20 g collagen protein respectively.The nerve conduits was crosslinked with genipin after freezing dehydration by gradient technique and sterilized irradiation with 60Co.The mechanical characteristics of collagen nerve conduits were evaluated with omnipotent material machine.The spatial structure of collagen nerve conduits before and after crosslinked were observed by scanning electron microscope（SEM）.Mesenchymal stem cells（MSC） mixed ectocellular mesenchyme（ECM） glue were inoculated in the conduit,the compound state of cell and material were studied in SEM.The MSC were induced 10 g/L ligustrazine for 7 days and differentiated into neuronal cells with immunochemistry technique.Eight Wister rats were anesthetized with 3 % pentobarbital sodium,exposed sciatic nerve and duplicated defect model.The collagen nerve conduits that contained MSC linked sciatic nerve,4 non-ligustrazine collagen nerve conduits as control,the rats were sacrificed after 90 days,nerve regeneration was observed and identified its function in immuno-histochemistry.Results The structure of collagen fiber was significantly changed,which arranged closely,formed irregular pore space and strengthened its intensity before and after crosslinked with genipin.The mechanical results of before and after crosslinked showed that the maximal load and fracture load of collagen nerve conduit were（0.23 ± 0.09） N,（0.20 ± 0.12） N and（0.76 ± 0.15） N,（0.69 ± 0.17） N respectively with significant difference（P ﹤0.01）.The ligustrazine could be promoted neuron specific enolase（NSE） expression in MSC and induced MSC differentiation into nerve cells.The collagen nerve conduits had the favourable hist

关 键 词：胶原蛋白 神经导管 京尼平 川芎嗪 神经缺损修复 

分 类 号：R651.3[医药卫生—外科学]