CAJ | 학술논문

[目的] 研究多次经蛛网膜下腔移植骨髓基质干细胞对Wistar大鼠脊髓损伤的功能修复作用.[方法] 骨髓基质干细胞经体外分离、培养并用Hoecst33342标记.按照Allen方法把60只在T_(10)-T_(12)平面损伤的Wistar大鼠随机分4组,A、B、C、D组(对照组).在损伤平面蛛网膜下腔中段放置一硅胶管,在7 d后,注入1×10~6个骨髓基质干细胞.在2、3、5、7、12周荧光显微镜、免疫组化检测骨髓基质干细胞在损伤段脊髓的存活、分布、分化情况并作计数观察.使用BBB评分观测后肢功能恢复.[结果] 移植后7～14 d骨髓基质干细胞达到高峰,在7 d后表达巢蛋白及神经丝蛋白阳性.随时间的延长,移植在损伤部位的骨髓基质干细胞数量及神经元样细胞均有减少,但移植3次的细胞数减少速度较其他两组慢.移植3次组大鼠的BBB评分较移植1、2次组有明显的提高,P＜0.01,有统计学意义.[结论] 多次移植骨髓基质干细胞更有利于脊髓损伤的恢复.
[목적] 연구다차경주망막하강이식골수기질간세포대Wistar대서척수손상적공능수복작용.[방법] 골수기질간세포경체외분리、배양병용Hoecst33342표기.안조Allen방법파60지재T_(10)-T_(12)평면손상적Wistar대서수궤분4조,A、B、C、D조(대조조).재손상평면주망막하강중단방치일규효관,재7 d후,주입1×10~6개골수기질간세포.재2、3、5、7、12주형광현미경、면역조화검측골수기질간세포재손상단척수적존활、분포、분화정황병작계수관찰.사용BBB평분관측후지공능회복.[결과] 이식후7～14 d골수기질간세포체도고봉,재7 d후표체소단백급신경사단백양성.수시간적연장,이식재손상부위적골수기질간세포수량급신경원양세포균유감소,단이식3차적세포수감소속도교기타량조만.이식3차조대서적BBB평분교이식1、2차조유명현적제고,P＜0.01,유통계학의의.[결론] 다차이식골수기질간세포경유리우척수손상적회복.
[Objective]To investigate the effect of multiple subarachnoid infusion of bone marrow stromal cells(BMSCs) on spinal cord injury repair in adult Wistar rats.[Method]The BMSCs were cultured, purified and labeled with Hoecst33342 in vitro. After the complete spinal cord injury rat model at T_(10)T_(11) level was successfully established, sixty injured rats were randomly divided into 4 groups: groups A, B, C, and D. The labeled BMSCs in a dose of 1×10~6 /0.2 ml were injected into subarachnoid space of the rats via a catheters under the aubarachnoid space in group A,B and C. The cells were administered only once in group A 1-week after injury. However, they were injected after injury 1 and 3 weeks in group B and 1, 3 and 5 weeks after injury in group C, respectively. Neurological functions were evaluated using the Basso-Beattie-Bresnahan (BBB) Scale 1, 3,7,9 and 12 weeks after transplantation. The immunohistochemistry and fluorescence microscopy were analyzed to observe the migration, survival, differentiation and histomorphology of BMSCs. [Result]The transplanted BMSCs survived and grew in the injured region and the nearby spinal cord. Some of them expressed pre-neuronal marker nestine, and others expressed neuronal marker neurofilament-200 (NF-200) 1 or 2 weeks after transplantation. As time went on, the planted cells gradually decreased. There was a significant improvement in the BBB scores and neurological functions of rats treated with BMSCs compared to those of rats in the control group (P<0.05). Furthermore, the three BMSC-treated groups achieved more functional recovery than the other BMSC-treated and control groups.[Conclusion]The study showed that multiple transplantations of BMSCs into subarachnoid space are beneficial, to some extend, to the repair of SCI. So this way may be a helpful method in restoring neurological function.