载硫酸软骨素蛋白多糖神经套管阻止大鼠坐骨神经缝接口处再生轴突逃逸  

CSPGs loaded in a gelatin sleeve prevent regenerated axons escaping from the sciatic nerve suture after nerve transection in rats

作　　者：何富林 邱帅 顾凡彬 刘小林[1] 朱庆棠[1] HE Fulin;QIU Shuai;GU Fanbin;LIU Xiaolin;ZHU Qingtang(Department of Microsurgery and Orthopedic Trauma,The First Affiliated Hospital of Sun Yat-sen University.Center for Peripheral Nerve Tissue Engineering and Technology Research.Guangdong Province Engineering Laboratory for Soft Tissue Biofabrication,Guangzhou 510080,China)

机构地区：[1]中山大学附属第一医院显微创伤手外科、广东省周围神经组织工程技术研究中心、广东省软组织生物制造工程实验室,广州510080

出　　处：《中华显微外科杂志》2020年第4期361-367,共7页Chinese Journal of Microsurgery

基　　金：国家自然科学基金(31670986);国家重点研发计划(2016YFC1101603);广东省科技计划项目(2014B020227001,2017A050501017);广州市科技计划项目(201807010082,201704030041)。

摘　　要：目的观察载硫酸软骨素蛋白多糖(CSPGs)神经套管在SD大鼠坐骨神经横断伤端-端缝合口处的作用,并进一步观察其对周围神经损伤后修复的效果。方法利用明胶作为载体构建载CSPGs神经套管,用于保护SD大鼠坐骨神经横断伤端端缝合口。2019年7月至9月,24只SD大鼠随机分为直接缝合组、空白组(无CSPGs的明胶套管)和套管组(载CSPGs套管),每组8只。术后5周时,每组3只大鼠行荧光金注射以标志术侧背根神经节的感觉神经元,其余5只大鼠于术后6周行电生理学检测后分别取材,用于神经组织的苏木精-伊红(HE)染色及镀银染色,同时取术侧腓肠肌行Masson染色,评估缝合口处坐骨神经的再生情况及神经损伤后再生修复效果。采用单因素方差分析进行数据分析,如果组间差异有统计学意义,则进一步采用LSD法进行两两比较,P<0.05为差异有统计学意义。结果组织学结果表明,在坐骨神经端-端缝合口处,再生神经纤维均有不同程度的紊乱再生及逃逸现象,直接缝合组、空白组和套管组再生神经纤维的逃逸距离分别是(787.19±213.77)μm、(547.17±167.71)μm和(350.60±68.58)μm,通过缝合口进入远端基底膜管的再生轴突数目分别是(6360±736.89)个/mm^2、(8040±673.05)个/mm^2和(9000±644.20)个/mm^2,术侧背根神经节中被荧光金标记的感觉神经元数目分别是(124.35±25.88)个/mm^2、(165.36±30.74)个/mm^2和(208.98±20.51)个/mm^2,套管组与另外两组相比,差异均有统计学意义(P<0.05);电生理检测及术侧腓肠肌的组织学切片均显示套管组神经损伤后再生修复效果比另外两组更好(P<0.05)。结论载CSPGs神经套管可通过CSPGs对再生轴突的抑制作用,有效减少再生神经纤维在坐骨神经横断伤端-端缝合口处的逃逸及紊乱再生,从而促进大鼠坐骨神经损伤后有效再生。Objective To observe the effect that covering the nerve suture with CSPGs-gelatin sleeve after the sciatic nerve transection in SD rats,and evaluate its effect to improve nerve regeneration in peripheral nerve transection model.Methods Covered and protected the sciatic nerve by end-to-end suture in the SD rats with CSPGs-gelatin sleeve.From July,2019 to September,2019,24 SD rats were randomly and evenly divided into 3 groups,which were direct suture group,gelatin sleeve group without CSPGs(blank group),and CSPGs-gelatin sleeve group(CSPGs group).In each group,3 rats were used to mark fluorescent gold at 5 weeks after operation.At 6 weeks after operation,histological and electrophysiological tests were performed to evaluate the tissue regeneration in the end to end suture and the effect of peripheral nerve regeneration after transection in the other 5 rats.One-way ANOVA was used for data analysis.If the difference between groups was statistically significant,LSD method would be further used for pairwise comparison.P<0.05 was considered statistically significant.Results The escape distances in direct suture groups,blank group and CSPGs group groups were(787.19±213.77)μm,(547.17±167.71)μm and(350.60±68.58)μm,respectively;The numbers of the axons that grow into the distal basement membrane tube in 3 groups were(6360±736.89)/mm^2,(8040±673.05)/mm^2 and(9000±644.20)/mm^2,respectively;The numbers of sensory nerve cells that were marked by fluorescent gold in the dorsal root ganglion were(124.35±25.88)/mm^2,(165.36±30.74)/mm^2 and(208.98±20.51)/mm^2 in 3 groups,respectively.The differences were significant compared with the CSPGs-gelatin sleeve group(P<0.05).Thus,it led to a better nerve regeneration after transection in CSPGs-gelatin sleeve group according to the electrophysiological test and histological section observation of gastrocnemius muscles(P<0.05).Conclusion CSPGs loaded in the gelatin sleeve can inhibit axons regeneration,and prevent the regenerated axons escaping from the end to end suture that m

关 键 词：硫酸软骨素蛋白多糖 坐骨神经损伤 再生 修复 神经套管 大鼠 

分 类 号：R688[医药卫生—骨科学]