中华骨科杂志
中華骨科雜誌
중화골과잡지
CHINESE JOURNAL OF ORTHOPAEDICS
2009年
10期
969-973
,共5页
班德翔%冯世庆%宁广智%刘洋%陈家童%郭世绂
班德翔%馮世慶%寧廣智%劉洋%陳傢童%郭世紱
반덕상%풍세경%저엄지%류양%진가동%곽세불
许旺细胞%脊髓损伤%神经生长因子%轴突
許旺細胞%脊髓損傷%神經生長因子%軸突
허왕세포%척수손상%신경생장인자%축돌
Schwann cells%Spinal cord injuries%Nerve growth factors%Axon
目的 通过不同途径移植自体激活雪旺细胞(autologous activated Schwann cells,AASCs),评价各种移植途径在修复脊髓损伤中的作用.方法 结扎Wistar大鼠双侧隐神经,1周后取下结扎处远端神经,在体外分离、培养、传代、纯化和鉴定后获得供实验用AASCs.60只Wistar大鼠制成T_(10)脊髓损伤模型后随机分为三组,1周后将预先用Hoechst33342标记的AASCs移植到三组大鼠体内:Ⅰ组,尾静脉移植;Ⅱ组,鞘内移植(经蛛网膜下腔);Ⅲ组,局部损伤处移植.术后采用BBB评分评价大鼠功能恢复.3个月后行BDA皮质脊髓束顺行示踪标记.标记2周后处死动物,取出损伤处脊髓行快速冰冻切片,行Cy3荧光探针染色、神经丝蛋白200(NF200)和HE染色.结果 AASCs在体外可稳定传4代以上,并表达S-100抗原.从术后第4周开始,BBB评分在各组间差异有统计学意义.至实验结束时,HE染色显示Ⅲ组中损伤空洞明显小于其余两组,NF200免疫组化染色阳性面积占总面积百分比各组间差异有统计学意义.BDA神经示踪显示,Ⅲ组中有较多的再生轴突通过脊髓损伤区,横断面上再生轴突的免疫组化阳性面积各组间差异有统计学意义.结论 局部损伤处移植AASCs可以有效保证移植细胞的数量,AASCs通过分泌多种营养因子和桥接损伤轴突再生的作用促进大鼠脊髓损伤后的功能恢复.
目的 通過不同途徑移植自體激活雪旺細胞(autologous activated Schwann cells,AASCs),評價各種移植途徑在脩複脊髓損傷中的作用.方法 結扎Wistar大鼠雙側隱神經,1週後取下結扎處遠耑神經,在體外分離、培養、傳代、純化和鑒定後穫得供實驗用AASCs.60隻Wistar大鼠製成T_(10)脊髓損傷模型後隨機分為三組,1週後將預先用Hoechst33342標記的AASCs移植到三組大鼠體內:Ⅰ組,尾靜脈移植;Ⅱ組,鞘內移植(經蛛網膜下腔);Ⅲ組,跼部損傷處移植.術後採用BBB評分評價大鼠功能恢複.3箇月後行BDA皮質脊髓束順行示蹤標記.標記2週後處死動物,取齣損傷處脊髓行快速冰凍切片,行Cy3熒光探針染色、神經絲蛋白200(NF200)和HE染色.結果 AASCs在體外可穩定傳4代以上,併錶達S-100抗原.從術後第4週開始,BBB評分在各組間差異有統計學意義.至實驗結束時,HE染色顯示Ⅲ組中損傷空洞明顯小于其餘兩組,NF200免疫組化染色暘性麵積佔總麵積百分比各組間差異有統計學意義.BDA神經示蹤顯示,Ⅲ組中有較多的再生軸突通過脊髓損傷區,橫斷麵上再生軸突的免疫組化暘性麵積各組間差異有統計學意義.結論 跼部損傷處移植AASCs可以有效保證移植細胞的數量,AASCs通過分泌多種營養因子和橋接損傷軸突再生的作用促進大鼠脊髓損傷後的功能恢複.
목적 통과불동도경이식자체격활설왕세포(autologous activated Schwann cells,AASCs),평개각충이식도경재수복척수손상중적작용.방법 결찰Wistar대서쌍측은신경,1주후취하결찰처원단신경,재체외분리、배양、전대、순화화감정후획득공실험용AASCs.60지Wistar대서제성T_(10)척수손상모형후수궤분위삼조,1주후장예선용Hoechst33342표기적AASCs이식도삼조대서체내:Ⅰ조,미정맥이식;Ⅱ조,초내이식(경주망막하강);Ⅲ조,국부손상처이식.술후채용BBB평분평개대서공능회복.3개월후행BDA피질척수속순행시종표기.표기2주후처사동물,취출손상처척수행쾌속빙동절편,행Cy3형광탐침염색、신경사단백200(NF200)화HE염색.결과 AASCs재체외가은정전4대이상,병표체S-100항원.종술후제4주개시,BBB평분재각조간차이유통계학의의.지실험결속시,HE염색현시Ⅲ조중손상공동명현소우기여량조,NF200면역조화염색양성면적점총면적백분비각조간차이유통계학의의.BDA신경시종현시,Ⅲ조중유교다적재생축돌통과척수손상구,횡단면상재생축돌적면역조화양성면적각조간차이유통계학의의.결론 국부손상처이식AASCs가이유효보증이식세포적수량,AASCs통과분비다충영양인자화교접손상축돌재생적작용촉진대서척수손상후적공능회복.
Objective To explore the curative effect of autologous activated Schwann cells (AASCs) in rat spinal cord injury (SCI) via different grafting routes. Methods Saphenous nerve of the adult Wistar rat was ligated for one week to activate Schwann cells. And then AASCs were cultured, passaged, purified and identified in vitro. Sixty Wistar rats were all made into SCI model at T_(10) and then divided into three groups randomly. One week after injury, AASCs were transplanted into the SCI rats via three different routes [group Ⅰ: tail vein; group Ⅱ: intrathecal (subarachnoid route); group Ⅲ: intraspinal cord]. BBB score was carried out to evaluate the functional recovery of the injured rats at weekly interval postinjury. Three months later, corticospinal tract (CST) was labeled by 10% BDA. Two weeks after labeling, the rats were sacrificed and then their injured spinal cord tissues were take out to carry out 10 μm fast frozen section, which was then followed by Cy3 fluorescent probe staining, NF200 and HE staining. Results AASCs passed over 4 stable passages and expressed S-100 antigen in vitro. The difference among groups could be considered sig-nificant in BBB score on the 5th week postinjury. HE staining showed that the injured cavity was evidently smaller in group Ⅲ compared with other two groups. Statistical difference also existed among groups by com-paring the percentage of positive response areas in NF200 staining. 10%BDA stain demonstrated that more regenerated axons passed through the injured cavity in group Ⅲ when compared with other two groups, which was also confirmed by immunohistochemistry stain. Conclusion The amount of grafted AASCs can be guaranteed effectively in injured epicenter via the grafting route of intraspinal cord. AASCs promote the repair after SCI by secreting multiple kinds of neurotrophic factors and bridge injured axons for regrowth.