经嗅鞘细胞条件培养液诱导的骨髓基质干细胞移植对脊髓损伤修复的影响  

Effects of transplantation of bone marrow stromal cells induced byolfactory ensheathing cells' conditioned medium on repair of spinal cord injury in rats

作　　者：冯林杰[1] 甘洪全[1] 刘英杰[1] 余向前[1] 刘昊[1] 毕成[1] 罗卓荆[2] 

机构地区：[1]河北联合大学附属医院骨科,唐山063000 [2]第四军医大学西京医院骨科

出　　处：《中国矫形外科杂志》2015年第18期1703-1709,共7页Orthopedic Journal of China

基　　金：河北联合大学科学研究基金项目(编号:Z201432);河北省医学科学研究课题计划项目(编号:20150508)

摘　　要：[目的]研究经嗅鞘细胞条件培养液诱导的骨髓基质干细胞(BMSCs)移植对脊髓损伤的治疗作用。[方法]采用脊髓挤压损伤方法,制造SD大鼠脊髓损伤动物模型(共24只),随机分成A、B、C三组(每组8只),立即分别将DMEM、BMSCs和经诱导的BMSCs移植入脊髓损伤部位,分别于移植4、8周后处死动物,均采用HE染色观察脊髓损伤处空洞面积改变情况;并采用抗神经丝(NF)抗体、生长相关蛋白-43(GAP-43)抗体和神经元特异性核蛋白(Neu N)抗体免疫组织化学荧光染色,观察移植的BMSCs存活和分化情况及损伤部位神经纤维再生情况。[结果]移植4、8周后,脊髓损伤区空洞为移植细胞充填,空洞面积明显减少,差异有统计学意义(P<0.01),BBB评分结果 C组和A、B组之间存在统计学差异(P<0.05)。B、C组均可见再生神经纤维形成,同时部分移植细胞呈Neu N染色阳性,在C组上述改变更加显著。[结论]经嗅鞘细胞条件培养液诱导的BMSCs可以在脊髓损伤区存活,能够显著减小脊髓损伤处空洞面积,并可以促进脊髓损伤区神经纤维的修复再生,促进大鼠双下肢运动功能的恢复。[ Objective] To investigate the effect of bone marrow stromal cells （BMSCs） induced byolfactory ensheathing cells （OECs） conditioned medium on repair of spinal cord injury in rats. [ Method ] Spinal cords of adult rats （n = 24） were in- jured by spinal cord crushing method, and immediately, the injured cords were injected with Dubecco -modified Eagles Medium （DMEM）, BMSCs and BMSCs which induced byolfaetory ensheathing cells ~ conditioned medium respectively. Rats were ob- served by Basso - Beatie - Bresnahan （BBB） scale from 1 week after injury, and all rats were killed at four weeks and eight weeks after operations. The lesioned spinal cord segments were removed, sectioned horizontally, stained with hematoxylin - eosin （HE） and immunostained with antibodies against neurofilament （NF）, growth associated protein- 43 （GAP- 43 ） and neuronal specificity nucleoprotein （ Neu N）. [ Result ] Four weeks and eight weeks after transplantation, significant reduction in lesion area （P 〈 0.01 ） and improvement in BBB locomotor rating scale （P 〈 0.05 ） were observed in the cases that received implantation of BMSCs which was induced byolfactory ensheathing cells＇conditioned medium. Immunocytochemical staining showed that trans- planted cells could survive in lesion area and partially expressed NeuN. While numerous NF and GAP -43 positive axonals were detected in lesion area of transplanted groups, and more were found in groups which was transplanted with BMSCs induced byol- factory ensheathing cells conditioned medium. [ Conclusion ] The transplanted BMSCs induced by ensheathing cells （OECs） conditioned medium can survive and reduce lesion size, promote axonal regeneration and improve functional recovery.

关 键 词：脊髓损伤 骨髓基质干细胞 条件培养液 移植 大鼠 

分 类 号：R651.2[医药卫生—外科学]