低温保存嗅神经鞘细胞移植对脊髓轴突再生的影响  

The influence of transplantation of cryopreserved olfactory ensheathingc ells on spinal cord axon regeneration in adult rats

作　　者：沈慧勇[1] 吴燕峰[2] 唐勇[1] 陈燕涛[1] 程志安[1] 

机构地区：[1]中山大学第二附属医院骨科,广州510120 [2]中山大学第二附属医院岭南脐血库,广州510120

出　　处：《中华骨科杂志》2002年第9期556-560,共5页Chinese Journal of Orthopaedics

基　　金：广东省科委重点攻关项目(C30702)

摘　　要：目的观察低温保存(-120℃)的嗅神经鞘细胞(OECs)移植对脊髓轴突再生及神经功能恢复的促进作用。方法将分离培养的SD大鼠OECs冻存2周,复温后移植于12只成年SD大鼠T 10水平SCI模型的脊髓两断端,另12只对照组SD大鼠只注入DMEM。分别于移植后6、12周行下肢功能评价、MEP检测、组织学和免疫组织化学检查。结果实验组及对照组动物存活率均为75%。移植后6周,实验组及对照组动物下肢瘫痪,下肢记录不到MEP。组织学检查(每组各2只)见实验组OECs与宿主融合良好,细胞内及细胞周围有髓鞘碱性蛋白阳性物质存在,宿主再生轴突经近侧断端长入断端间组织;对照组远、近断端轴突变性,未见再生轴突。移植后12周,实验组2只动物(2/7)下肢出现肌肉收缩,2只(2/7)髋、膝关节能活动,5只记录到MEP;对照组动物(7/7)下肢功能无改善,记录不到MEP。组织学检查见实验组OECs明显减少,部分再生轴突穿过损伤区组织抵达脊髓远侧断端;对照组瘢痕形成,无轴突再生。结论低温保存的OECs移植后可在脊髓中存活,并产生髓鞘样物质,促进宿主脊髓轴突再生及神经功能部分恢复。Objective To observe the effects of cryopreserved olfactory ensheathing cells(OECs )transplantation on spinal cord axonal regeneration and spinal func tional improvement.Methods Twenty-four rats were divided into experimental a nd control groups,with each group of 12rats.The spinal cord in-jury was es tablished by transecting the spinal cord with microsurgery scissors.OECs were p urified from SD rat olfactory bulb and cultured in DMEM and cryopreserved(-1 20℃)for 2weeks.OECs suspension[(1～1.4)×10 5 /ml ]was transplanted into transected spinal cord at T 10 level,while the DMEM solution was in ject-ed instead in the control group .At 6weeks and12weeks after transplantation,the rats were evaluated with cl imbing test and MEP monitoring.The samples of spinal cord were procured and st udied with histological and immunohistochemical measurement.Results At 6weeks after transplantation,all of the rats in both transplanted and control group were paraplegic,and MEPs could not be recorded.Morphol ogy of trans-plante d OECs was normal,and OECs were interfused with host well.Axons could regrow i nto gap tis sue be-tween the spinal cords.Both OECs and regrowed axons were im munoreactive for MBP.No re grown axons were found in the control group.At 12weeks after transplantation,2rats(2/7)had lower ex tremities mus-cle contraction,2rats(2/7)had hip and /or knee active movement,and MEP of 5rats(5/7)could be recorded in the calf in the transplantation group.Non e of the rates(7/7)in the control group had functional improvement,and n one of them had MEPs recorded.In the transplanted group,histological and imm unohis-tochemical methods showed the number of transplanted OECs reduced and s ome regrown axons had reached the end of transected spinal cord.However,no r egrown axons could be seen except scar forma tion in the control group.Concl usion Cryopreserved OECs could integrate with the host and promote re growing a x-ons across the transected spinal cord ends.

关 键 词：嗅神经 脊髓损伤 轴突 再生 低温 

分 类 号：R651.2[医药卫生—外科学]