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.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
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)
关键词
脊髓损伤
骨髓基质干细胞
条件培养液
移植
大鼠
spinal cord injury, bone marrow stromal cells, conditioned medium, transplantation, rats
