CAJ | 학술논문

背景:目前细胞移植是修复脊髓损伤的研究热点之一,许旺细胞能够分泌多种神经营养因子,改善损伤局部微环境,大量相关文献证实了其能够促进脊髓损伤后功能的恢复.针对治疗脊髓损伤的细胞移植方法很多,其中静脉移植具有操作方便、能够避免传统移植方法造成的附加损伤等优点.目的:探讨许旺细胞尾静脉移植修复大鼠脊髓损伤的疗效.方法:体外分离Wistar大鼠双侧坐骨神经,植块法培养,S-100染色鉴定,Hoechst33342标记许旺细胞.Impactor model-Ⅱ打击仪制作 Wistar大鼠T10脊髓损伤模犁60只,随机分为3组,空白对照组术后未作处理,DMEM对照组、许旺细胞移植组术后1周分别尾静脉注射1 mL.DMEM或1 mL.许旺细胞.术前1 d,术后1,3 d,1周及以后每周进行BBB评分.移植后1,2,4周取材行冰冻切片观察移植许旺细胞的迁移.移植后8周取材行苏木精-伊红染色及免疫荧光染色观察损伤段胶质原纤维酸性蛋白、神经元特异性烯醇化酶的表达.结果与结论:经纯化鉴定获得纯度为95%的许旺细胞.移植后1,2,4周损伤段及邻近节段脊髓内可观察到Hoechst33342阳性细胞.术后4周,许旺细胞移植组与空白对照组、DMEM对照组BBB评分差异开始有显著性意义(P<0.05),且许旺细胞移植组高于空白对照组、DMEM对照组.苏木精-伊红染色示各组均有脊髓空洞形成,许旺细胞移植组空洞小于空白对照组、DMEM对照组.免疫组织化学染色示空白对照组、DMEM对照组胶质原纤维酸性蛋白反应较强,许旺细胞移植组胶质原纤维酸性蛋白阳性面积小于其他组(P<0.05);许旺细胞移植组神经元特异性烯醇化酶的阳性面积明显大于其他组(P<0.05).提示静脉移植许旺细胞修复大鼠脊髓损伤是可行的,操作简便,且有较好的疗效.
배경:목전세포이식시수복척수손상적연구열점지일,허왕세포능구분비다충신경영양인자,개선손상국부미배경,대량상관문헌증실료기능구촉진척수손상후공능적회복.침대치료척수손상적세포이식방법흔다,기중정맥이식구유조작방편、능구피면전통이식방법조성적부가손상등우점.목적:탐토허왕세포미정맥이식수복대서척수손상적료효.방법:체외분리Wistar대서쌍측좌골신경,식괴법배양,S-100염색감정,Hoechst33342표기허왕세포.Impactor model-Ⅱ타격의제작 Wistar대서T10척수손상모리60지,수궤분위3조,공백대조조술후미작처리,DMEM대조조、허왕세포이식조술후1주분별미정맥주사1 mL.DMEM혹1 mL.허왕세포.술전1 d,술후1,3 d,1주급이후매주진행BBB평분.이식후1,2,4주취재행빙동절편관찰이식허왕세포적천이.이식후8주취재행소목정-이홍염색급면역형광염색관찰손상단효질원섬유산성단백、신경원특이성희순화매적표체.결과여결론:경순화감정획득순도위95%적허왕세포.이식후1,2,4주손상단급린근절단척수내가관찰도Hoechst33342양성세포.술후4주,허왕세포이식조여공백대조조、DMEM대조조BBB평분차이개시유현저성의의(P<0.05),차허왕세포이식조고우공백대조조、DMEM대조조.소목정-이홍염색시각조균유척수공동형성,허왕세포이식조공동소우공백대조조、DMEM대조조.면역조직화학염색시공백대조조、DMEM대조조효질원섬유산성단백반응교강,허왕세포이식조효질원섬유산성단백양성면적소우기타조(P<0.05);허왕세포이식조신경원특이성희순화매적양성면적명현대우기타조(P<0.05).제시정맥이식허왕세포수복대서척수손상시가행적,조작간편,차유교호적료효.
BACKGROUND: Emerging studies have focused on cell transplantation. Schwann cells (SCs) can secrete various neurotrophic factors and improve local environment around injury. Plenty of documents have demonstrated that SCs could promote functional recovery following spinal injury. Many transplanting methods are available for treating spinal cord injury, and the intravenous cell transplantation is profitable for easy operation and avoidance of additional trauma. OBJECTIVE: To investigate the effects of intravenous transplantation of SCs on spinal cord injury in rats. METHODS: The bilateral sciatic nerves of Wistar rats were separated in vitro, cultured by tissue clot method, identified by S-100 and labeled by Hoechst33342. Sixty rat models with T10 spinal cord injury were prepared using impactor model- II type weight drop apparatus. Then the injured rats were randomly divided into 3 groups: blank control, DMEM control and SCs transplantation groups. No treatment was performed in the blank control group. Totally 1 mL DMEM and or SCs was injected into rats of DMEM control and SCs transplantation groups by tail vein respectively. Basso Beattie Bresnahan (B6B) scores were performed at 1 day before and 1, 3 days, 1 week and weekly after operation. The migration of transplanted SCs was observed at 2 weeks and 4 after transplantation. The expressions of glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were detected by haematoxylin-eosin staining and immune-fluorescence staining.RESULTS AND CONCLUSION: The purity of SCs reached 95%. Hoechst33342 positive cells were observed throughout the injured and the nearby region of spinal cord at 1, 2, and 4 weeks after transplantation. The statistical difference of BBB score among the SCs transplantation, blank control, and the DMEM control groups displayed at 4 weeks after transplantation (P < 0.05), and the BBB scores of the SCs transplantation were higher than other groups. Haematoxylin-eosin staining showed the cavity formed in each group at 8 weeks after transplantation, but the area of SCs transplantation was smaller than that of the blank control and DMEM control groups. The immunofluorescence staining indicated that the expression of GFAP were more intense in the blank control group and DMEM control than SCs transplantation (P < 0.05), while the expression of NSE was more intense in SCs transplantation than other groups (P< 0.05). It implied that intravenous transplantation of SCs promotes regeneration of axon and improves neurological functions after spinal cord injury in rats.