NT-3修饰的施万细胞联合胶原—壳聚糖神经导管对大鼠坐骨神经损伤的修复作用研究  

Repairing effect of NT-3-modified Schwann cells combined with collagen-chitosan nerve conduit on sciatic nerve injury in rats

作　　者：许蕙[1] 叶放[1] 于宁[1] 王艳生 XU Hui;YE Fang;YU Ning;WANG Yan-sheng(Department of Hand Surgery,Affiliated Central Hospital of Shenyang Medical College,Shenyang Liaoning 110024,China)

机构地区：[1]沈阳医学院附属中心医院手外科,辽宁沈阳110024

出　　处：《局解手术学杂志》2021年第1期1-6,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：辽宁省自然科学基金指导计划项目(20180550853)。

摘　　要：目的观察神经营养素3(NT-3)修饰的施万细胞(SCs)联合胶原—壳聚糖神经导管对大鼠坐骨神经损伤的修复作用。方法将75只SD大鼠随机分为对照组(A组,n=12),模型组(B组,n=10),神经导管单独移植组(C组,n=11),未修饰施万细胞+神经导管移植组(D组,n=20),NT-3修饰施万细胞+神经导管移植组(E组,n=22)。各组大鼠麻醉后,暴露左侧坐骨神经,切除8 mm神经干制备坐骨神经损伤模型,然后分别给予自身坐骨神经翻转吻合后缝合、切除坐骨神经后缝合、神经导管桥接缝合缺损神经、未修饰的施万细胞复合神经导管桥接缝合缺损神经和NT-3修饰的施万细胞复合神经导管桥接缝合缺损神经。S100/Sox10双荧光染色法鉴定从新生SD大鼠提取的施万细胞纯度,RFP标记腺病毒载体(AdvNT-3)确定最佳MOI值,Live/Dead染色法检测术后5 d、11 d移植处施万细胞存活状态,坐骨神经功能指数(SFI)法和电生理检测评估术后大鼠神经功能恢复情况,TUNEL测定神经元细胞凋亡情况,采用透射电镜观察坐骨神经髓鞘再生情况。结果从新生SD大鼠提取的施万细胞纯度为95%以上,NT-3胰腺病毒感染的最佳MOI值为100,且移植后施万细胞在胶原—壳聚糖神经导管中存活状态良好。术后1周,与A组比较,其余各组SFI均明显降低(P<0.05),但其余各组间比较差异无统计学意义(P>0.05);术后4周,与A组比较,其余各组SFI均显著降低(P<0.05),其余各组中E组最高(P<0.05),而B组、C组、D组间无统计学差异(P>0.05);术后7周、10周和13周,与B组和C组相比,D组和E组SFI均显著较高(P<0.05),且E组高于D组(P<0.05),B组和C组比较无统计学差异(P>0.05)。术后13周,与A组相比,其余各组动作电位传导速度显著较低(P<0.05),除E组外其余各组峰峰值均较低(P<0.05);与B组相比,C组、D组和E组动作电位传导速度和峰峰值依次升高(P<0.05)。术后13周,与其他组相比,B组神经元细胞凋亡数量明显较多,神经Objective To observe the repairing effect of neurotrophin 3(NT-3)-modified Schwann cells(SCs)combined with collagen-chitosan nerve conduit on sciatic nerve injury in rats.Methods A total of 75 SD rats were randomly divided into control group(group A,n=12),model group(group B,n=10),nerve conduit transplantation group(group C,n=11),unmodified Schwann cells+nerve conduit transplantation group(group D,n=20),and NT-3-modified Schwann cells+nerve conduit transplantation group(group E,n=22).After anesthesia,the left sciatic nerve was exposed and the sciatic nerve injury model was established by cutting 8 mm nerve trunk.Then they were given suture after reverse anastomosis of sciatic nerve,suture after resection of the sciatic nerve,suture bridged by nerve conduit,suture bridged by unmodified Schwann cells combined with nerve conduit,suture bridged by NT-3 modified Schwann cells and nerve conduit,respectively.S100/Sox10 double fluorescence staining method was used to identify the purity of Schwann cells from neonatal SD rats.The optimal MOI value was determined by using adenovirus vector(AdvNT-3)labeled with RFP.Live/Dead staining method was used to detect the survival status of the Schwann cells 5 days and 11 days after operation.Sciatic nerve function index(SFI)and electrophysiological examination were used to evaluate the recovery of neurological function.TUNEL was used to detect neuronal cell apoptosis.And the transmission electron microscope was used to observe the regeneration of myelin sheath of sciatic nerve.Results The purity of Schwann cells from neonatal SD rats was more than 95%.The optimal MOI value of NT-3 pancreatic virus infection was 100.After transplantation,Schwann cells grew well in collagen-chitosan neural conduit.Compared with group A,the SFI of the other groups were significantly reduced 1 week after operation(P<0.05),but there was no statistically significant difference among the other groups(P>0.05).Compared with group A,the SFI of the other groups were significantly reduced 4 weeks after operati

关 键 词：神经营养素3 施万细胞 坐骨神经损伤 胶原—壳聚糖神经导管 大鼠 动物模型 周围神经系统 

分 类 号：R285.5[医药卫生—中药学]