含FGL活性片段的自组装多肽支架材料修复大鼠脊髓损伤的研究  

Effect of functionalized peptide self-assembling nanofiber scaffolds containing FGL motif on rat spinal cord injury repair

作　　者：邹枕玮[1] 郝少飞[2] 郑启新[2] 吴永超[2] 郭晓东[2] 

机构地区：[1]华中科技大学同济医学院附属协和医院肿瘤中心,武汉430022 [2]华中科技大学同济医学院附属协和医院骨科,武汉430022

出　　处：《中华实验外科杂志》2013年第10期2178-2181,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30973027、81171159、81200313）

摘　　要：目的 建立大鼠脊髓损伤模型,将含FGL功能化多肽自组装神经支架材料（FGL-NS）植入到脊髓损伤局部,探讨FGL-NS神经支架材料修复脊髓损伤的效果和机制.方法 麻醉大鼠后,暴露胸10水平脊髓,用特制血管夹（夹力24g）钳夹1 min,建立大鼠胸髓钳夹损伤模型.设立空白组、对照组和实验组,分别于损伤后24h,暴露脊髓损伤局部,将2.5μl等渗葡萄糖溶液、1％ RADA-16和1％ FGL-NS多肽溶液注射入损伤局部.术后3d、1、3、5、7和9周分别采用Basso-Beattie-Bresnahan（BBB）评分评估大鼠脊髓损伤经治疗后运动功能恢复情况,术后9周取脊髓损伤部位组织,行半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3、神经丝蛋白-200 （NF-200）和胶质纤维酸性蛋白（GFAP）免疫组织化学染色评估损伤部位细胞凋亡、轴突再生和瘢痕形成情况.结果 成功建立大鼠胸髓钳夹损伤模型.BBB运动学评分大鼠脊髓损伤后经FGL-NS材料治疗后,BBB评分随时间逐渐升高,自伤后第5周起,显著高于RADA-16治疗组和空白组,大鼠运动功能显著改善.免疫组织化学染色结果显示,损伤后第9周,FGL-NS治疗组脊髓损伤局部可见大量NF-200阳性的神经元细胞[（35.32±3.12）个/视野],显著高于RADA-16组[（18.56±2.64）个/视野]和空白组[（14.83±1.43）个/视野],Caspase-3阳性的凋亡细胞[（22.45 ±2.74）个/视野]显著少于RADA-16组[（30.86 ±3.75）个/视野],而且损伤区GFAP染色的积分吸光度（IA）值为0.50±0.02,明显小于对照组（1.30 ±0.09）和空白组（1.60±0.11）.结论 功能化多肽自组装神经支架材料FGL-NS能促进脊髓损伤大鼠的运动功能恢复,能减少脊髓损伤部位细胞凋亡,促进神经元再生,并减少胶质瘢痕形成.Objective To establish a rat model of spinal cord injury （SCI） and evaluate the effect of in vivo treatment of SCI with FGL-NS peptide that could be self-assembled in vivo into functionalized nanofiber scaffold.Methods The rats were anesthetized and treated with laminectomy at the T10 vertebral segment.Then,the SCI models were established by compressing the spinal cord with a 24 g modified aneurysm clip.One day after SCI,the animals were respectively injected with 2.5 μl 580 μmol/L solution of glucose,1％ RADA-16 and 1％ FGL-NS peptide solution into the injured zone.After treatment for 3 days,1 week,3 weeks,5 weeks,7 weeks and 9 weeks,the functional recovery of SCI rats was evaluated by the Basso,Beattie and Bresnahan Locomotor Rating Scale.After treatment for 9 weeks,apoptosis rate and the number of neurons with neurite and astrocyte were detected by Caspase-3,neurofilament-200 （NF-200） and glial fibrillary acidic protein （GFAP） immunohistochemistry.Results The model of SCI in rats was established successfully.The Basso-Beattie-Bresnahan （BBB） scores of SCI rats treated with FGL-NS were higher than those in RADA-16 group after treatment for 5 weeks.The number of Caspase-3 positive cells in injuried zone treated with FGL-NS was （22.45 ± 2.74） per field,which was significantly less than in RADA-16 group [（30.86 ±3.75） per field].Meanwhile,the number of neurons with neurite in FGL-NS group was （35.32 ±3.12） per field,significantly more than that in RADA-16 group [（18.56 ±2.64） per field] and blank group [（14.83 ± 1.43） per field].Furthormore,FGL-NS inhibited the formation of astrogila at the site of injury.The integrated option density （IOD） in injuried zone treated with FGL-NS was 0.50 ± 0.02,significantly less than in control group （1.30 ± 0.09） and blank group （1.60 ± 0.11）.Conclusion Functionalized peptide self-assembling nanofiber scaffolds containing FGL motif could be a useful biomaterial in nerve tissue engineering for improving the functional reco

关 键 词：脊髓损伤 自组装 再生 组织工程 支架材料 

分 类 号：R318.08[医药卫生—生物医学工程]