显微操作在脐血干细胞移植修复大鼠脊髓损伤中的应用  

Micromanipulation used in surgical procedure of transected spinal cord treated with umbilical cord blood stem cells grafts in rats

作　　者：尹文化[1] 金大地[1] 鲁凯伍[1] 闫慧博[1] 王璐[1] 邓许勇[1] 

机构地区：[1]南方医科大学附属南方医院脊柱外科,广州510515

出　　处：《中国矫形外科杂志》2008年第24期1887-1890,共4页Orthopedic Journal of China

基　　金：国家自然科学基金(30471761);广东省自然科学基金(04020415)

摘　　要：[目的]建立大鼠脊髓全横断损伤模型,应用显微操作方法和人脐血干细胞移植修复大鼠脊髓损伤,观察显微操作技术对大鼠受损脊髓功能恢复的影响。[方法]取足月健康顺产新生儿脐血,分离、培养脐血干细胞。60只SD雌性大鼠,随机分为假手术对照组(A组,n=10)、实验对照组(B组,n=10)、常规治疗组(C组,n=20)和显微治疗组(D组,n=20)。A组大鼠只打开椎板;B、C组大鼠采用常规方法行T8~9平面脊髓全横断,造成急性脊髓损伤,C组大鼠于脊髓两断端用1μl注射器分别注射脐血干细胞悬液1μl(干细胞浓度为6×109/L^7×109/L),B组同法注射等量PBS液;D组采用显微操作技术行脊髓横断,并以口径50μm的毛细玻璃针用相同方法注射脐血干细胞。术后1~8周内每周进行1次后肢BBB评分,术后第8周处死大鼠,对比观察术中和术后第8周各组大鼠脊髓大体标本外形、颜色、质地和体积大小改变。[结果]A组大鼠手术前后运动功能、脊髓大体标本未见明显变化。B、C、D组大鼠术后双侧后肢完全性瘫痪,B组大鼠无明显恢复。而C、D组大鼠,从术后第2周开始逐渐恢复部分后肢运动功能。第3周以后,D组大鼠后肢运动功能BBB评分明显优于C组,具有显著性差异(P<0.05)。术后第8周,所有大鼠脊髓横断处均瘢痕愈合,B组大鼠远侧段脊髓干瘪、皱缩;C组大鼠远侧段脊髓直径轻度变细,外形尚饱满;D组大鼠横断处两侧脊髓直径无明显差异,外形圆润,但局部可见明显瘢痕粘连。[结论]人脐血干细胞移植对脊髓横断性损伤具有较好的治疗作用,显微操作技术的应用能最大可能地减少人为造成的脊髓损伤,有利于脊髓损伤的功能修复。[Objective] To establish effective and reliable complete spinal cord transection models in rats, and to investigate the effects of micromanipulation used in surgical procedure of transected spinal cord treated with human umbilical cord blood stem cells （UCBSCs） grafts in rats. [Method]Human UCBSCs were obtained from umbilical cord blood of term deliveries. Sixty Sprague-Dawley （SD） female rats were divided randomly into group A（ control group）, group B（ spinal cord transection and PBS injection）, group C （spinal cord transection and UCBSCs transplantation） and group D（spinal cord transection and UCBSCs trans- plantation by using micromanipulation）. All groups were operated, and 1μl UCBSCs（6×10^9/L-7×10^9/L） were injected into the both ends of the completely transected spinal cord at the groups C and D by two different surgical procedures respectively. From 1 week to 8 week post-operation, a behavioral testing was performed weekly upon each hindlimb for all animals according to the BBB scoring system. At the 8^th week, all animals were sacrificed and the spinal cords-were taken out for morphological obser-vation. [Result] No changes were observed at group A pre- and post-operation. After 2 weeks post-operation, the hindlirnb motor function of groups C and D began to recover. After 3 weeks post- operation, the group D showed more improvement than group C, and the difference between groups D and C were significant in the BBB scores （P〈0.05 ）. No functional recovery was observed at group B. In addition to the adhesion between the spinal dura matter and peripheral tissues at group D. hypertrophic scar existad in the injured spinal cord of every experimental rat. [Conclusion] Human UCBSCs may improve functional recovery of the hindlimbs of the experimental rats. Micromanipulation is benefit to elevating the therapeutic efficacy, because it might minimize the possibility of spinal cord injury in surgical procedure.

关 键 词：显微操作 脊髓损伤 脐血干细胞 移植 

分 类 号：R651.2[医药卫生—外科学]