外胚层间充质源神经干细胞-纤维蛋白支架复合体移植修复大鼠脊髓损伤  

Transplantation of the complex of ectomesenchymal derived neural stem cells and fibrin scaffold can repair the spinal cord injury in rats

作　　者：张雪松[1] 李正南[1] 荆丹峰 许艺荠 孙太存[1] 崔学文[1] 

机构地区：[1]江苏大学附属医院骨科

出　　处：《解剖学杂志》2016年第2期207-210,225,F0004,共6页Chinese Journal of Anatomy

摘　　要：目的：评价外胚层间充质来源的神经干细胞一纤维蛋白支架复合体移植治疗大鼠脊髓损伤的疗效。方法：于大鼠鼻呼吸黏膜组织获取外胚层间充质干细胞（EMSCs），并诱导为神经干细胞（NSCs）。制作大鼠脊髓横断模型，将EMSCs源性神经干细胞一纤维蛋白支架复合体移植入脊髓断端（作为NSCs-纤维蛋白支架移植组；N-F组），每周对大鼠行BBB评分，12周后对脊髓标本行免疫组织化学显色，并利用免疫印迹检测脊髓横断部位神经生长相关蛋白-43（GAP43）、神经丝蛋白-200（NF200）、突触蛋白（synapsin）的表达量，并与纤维蛋白支架移植组（Fg组）、单纯手术组（SCI组）进行比较。结果：鼻呼吸黏膜EMSCs源性神经干细胞高表达巢蛋白（nestin）、CDl33，在纤维蛋白胶内生长良好，并可形成神经元样突起。从术后2周开始，N-F组BBB评分每周均高于Fg组和SCI组，差异有统计学意义。免疫印迹检测显示N-F组动物脊髓标本NF200、GAP43、synapsin相对表达量均高于Fg组和SCI组。脊髓组织切片免疫组织化学显示脊髓断端N-F组可见大量阳性神经纤维呈网络状结构，并伸人断端两侧脊髓实质；Fg组有较少量纤维通过；SCI组几乎没有阳性纤维通过。结论：纤维蛋白支架对鼻呼吸黏膜EMSCs源性NSCs的生长分化具有良好的促进作用。EMSCs源NSCs对脊髓损伤后的组织学重建和功能恢复均具有明显的促进作用。Objective： To evaluate the curative effect of transplantation of the complex of ectomesenchymal derived neural stem cells and fibrin scaffold on repairing spinal cord injury in rats. Methods： The ectomesenchymal stem cells（EMSCs）were obtained from nasal respiratory mucosa in vivo of adult rats, cultured in vitro, and then induced into neural stem cells （NSCs）. The operation of transplanting the complex of EMSCs induced NSCs and fibrin scaffold to the lesion site of the spinal cord transection model in rats was performed （as the N-F group）, and then model rats were respectively estimated by Basso Beattie Bresnahan （BBB） locomotor rating score every week. The segments of spinal cord including the lesion site were harvested and examined by immunohistochemical staining. Expression of growth associated protein-43 （GAP43）, neurofilament-200 （NF200） and synapsin in the lesion site were performed by Western blotting. All the results were compared with the Fg group（single fibrin scaffold transplantation） and SCI group （simple operation）. Results： The EMSCs can be induced into NSCs with remarkable expression of nestin and CD133 examined by immunofluorescence. EMSCs derived NSCs reproduced well in fibrin glue and formed nerve-cell shaped projections. The movement function of the hind limbs in rats in three groups recovered respectively, and the BBB scores of rats in the N-F group were higher than that in the Fg and the SCI group at 2 weeks after operation. Western blotting showed that the expression of GAP43, NF200 and synapsin in the lesion site of the spinal cord in N-F group was higher than that in Fg and SCI group （P（0.05）. The immunohistochemistry of the spinal cord transection end in N-F group showed a lot of immunoreactive-positive nerve fibers with the form of network structures and the fibers extending into the broken end on both sides of the spinal cord parenchyma; less fiber was observed to pass through in Fg group; almost no positive fiber was observed to pass through

关 键 词：呼吸黏膜 外胚层间充质干细胞 脊髓损伤 神经干细胞 纤维蛋白胶 神经中继 

分 类 号：R651.2[医药卫生—外科学]