胶原-壳聚糖支架对脊髓损伤后运动功能恢复作用的实验研究  

Study on the effect of collagen-chitosan scaffolds on motor function recovery after spinal cord injury

作　　者：朱旭 于国渊[1] 杨华堂[1] 张宁[1] 王喜旺[1] 王晶[1] 王如科 ZHU Xu;YU Guoyuan;YANG Huatang;ZHANG Ning;WANG Xiwang;WANG Jing;WANG Ruke(Handan Central Hospital,Handan 056000,China)

机构地区：[1]邯郸市中心医院,河北邯郸056000

出　　处：《中国实用神经疾病杂志》2020年第23期2032-2038,共7页Chinese Journal of Practical Nervous Diseases

基　　金：邯郸市科学技术研究与发展计划项目(编号:16232080634)。

摘　　要：目的探讨胶原-壳聚糖支架对脊髓损伤(spinal cord injury,SCI)后运动功能恢复作用,为组织工程学在临床治疗脊髓损伤的应用奠定基础。方法(1)制备胶原与壳聚糖配比为31的凝胶复合材料,置于真空冻干机冷冻干燥得到多孔可降解的胶原-壳聚糖支架(scaffolds S);(2)选取成年雌性SD大鼠30只,体质量250~300 g,随机分为3组,暴露脊髓后未损伤脊髓为空白对照组(SHAM),采用挂线法离断脊髓后未放入支架为离断损伤组(SCI)和采用挂线法离断脊髓后植入胶原-壳聚糖支架为支架组(SCI+S);(3)术后每周采用双盲法通过Basso-Beattie-Bresnahan(BBB)评分评价各组大鼠后肢运动功能的恢复;分别在术后即刻、术后1个月和术后2个月分别检测各组大鼠(n=5)双后肢运动电生理(motor evoked potentials,MEP)活动;(4)术后8周取材,石蜡切片观察HE染色;冰冻切片观察神经丝蛋白(neurofilament,NF)免疫荧光染色情况。结果挂线法将缝合线穿过暴露的脊髓底部,提拉起脊髓后用手术剪刀离断,确保彻底离断脊髓组织。离断瞬间大鼠双下肢剧烈抽搐数次后各个关节松弛,无牵拉反射,提示造模成功。术后1个月和术后2个月,SCI+S组大鼠的MEP电生理活动、BBB评分结果均明显优于SCI组。HE染色结果发现SCI+S组损伤后形成的空洞明显少于SCI组大鼠;NF免疫荧光结果发现SCI+S组相对于SCI组更有利于脊髓损伤后的神经再生和修复。结论孔胶原-壳聚糖支架减轻脊髓损伤后空洞的形成并促进神经纤维及轴突再生及后肢运动功能恢复,是一种具有良好应用前景的治疗脊髓损伤的支架。Objective To investigate the effect of collagen-chitosan scaffold on motor function recovery after spinal cord injury(SCI),and to lay a foundation for the clinical application of tissue engineering in the treatment of spinal cord injury.Methods Collagen-chitosan composites with the ratio of 31 were prepared and frozen-dried in vacuum freeze-dryer to obtain porous scaffolds(S).Thirty adult female SD rats weighing 250-300 g were randomly divided into three groupscontrol group(SHAM),SCI group(SCI)and SCI+S group which were operated using thread-drawing method and then implanted collagen-chitosan scaffold immediately after spinal cord exposure.Basso-Beattie-Bresnahan(BBB)score was used to evaluate the recovery of hind limb motor function in each group every week after operation,and motor evoked potentials(MEP)activity was detected immediately after operation,1 month and 2 months after operation of each group respectively.After 8 weeks,the rats were sacrificed.The spinal cord were buried in paraffin section and stained with HE.The neurofilament NF immunofluorescence staining was observed by frozen section.Results Thread-drawing method ensured that the spinal cord was completely disconnected and the reliability of the model is ensured.At the moment of amputation,the muscles of the hind limbs of rats were flabby after severe convulsions,and there was no retraction reaction after pain stimulation,suggesting that the model of transected rats was successful.The results of MEP electrophysiological activity and BBB score in SCI+S group were significantly better than those in SCI group at 1 month and 2 months after operation.HE staining showed that the number of holes in SCI+S group was significantly less than that in SCI group;NF immunofluorescence showed that SCI+S group was more conducive to nerve regeneration and repair after spinal cord injury than SCI group.Conclusion Porous collagen-chitosan scaffold can alleviate the formation of cavity after spinal cord injury,promote the regeneration of nerve fibers and axons,and re

关 键 词：脊髓损伤 组织工程 胶原蛋白 壳聚糖 支架 运动功能恢复 

分 类 号：R-332[医药卫生]