摘要
目的在诱导椎间盘退变的早期移植同种异体兔髓核细胞(NPC),观察NPC对退变椎间盘的修复效果。方法体外分离、培养NPC,并通过组织化学染色、免疫组织化学和免疫荧光染色对其进行鉴定。采用穿刺抽吸法建立兔椎间盘退行性变模型,造模同期进行细胞移植治疗。将27只5月龄日本大耳白兔随机分为3组:正常对照组(n=9)、NPC移植组(NPC组,n=9)及DMEM培养基组(DMEM组,n=9)。每只实验动物的L 3-4、L 4-5和L 5-6作为实验干预椎间盘。术后4、8、12周分别利用标准化T2加权像信号强度(%ST2WI)、病理组织学、退变分数、免疫组化染色、免疫荧光染色以及二型胶原(CollagenⅡ)和蛋白聚糖(aggrecan)mRNA表达水平评价修复效果。结果术后12周,DMEM组的%ST2WI值、CollagenⅡ和aggrecan的mRNA表达量均明显低于其它两组(P<0.05),正常对照组和NPC组无明显差异(P>0.05);正常对照组和NPC组退变分数明显低于DMEM组(P<0.05),正常对照组和NPC组无明显差异(P>0.05);正常对照组、NPC组的CollagenⅡ免疫组化和aggrecan免疫荧光染色均显示强阳性,而DMEM组随着时间的推移阳性逐渐减弱。结论在退变早期移植同种异体的兔NPC能有效修复椎间盘组织的髓核退变。
Objective The aim of this study was to observe on the repair effect of rabbits’degenerative intervertebral disc on early stage with allogeneic nucleus pulposus cell(NPC)transplantation.Methods NPC was isolated and cultured in vitro,and identified with histochemical staining,immunochemical staining and Immunofluorescence staining.The intervertebral discs degeneration animal model was constructed by the conventional punctured method and at the same time NPC was transplanted into the rabbit’s model of disc degeneration.Twenty-seven 5-month-old Japanese white rabbits were randomly divided into 3 groups.Health intervertebral disc group was set as the normal control(NC)group(n=9).The NPC transplantation group was set as the positive group.Dulbecco’s modifed Eagle’s medium(DMEM)injection group was set as the negative group.L3-4、L4-5 and L5-6 intervertebral discs were set as samples of experiment in every rabbits.In 4,8 and 12 weeks after the transplantation,Repair effect of rabbits’degenerative intervertebral discs were evaluated by standard T2-weighted image ratio(%ST2WI),histology,degeneration score,immunochemical staining,Immunofluorescence staining,and mRNA expression levels of collagen II and aggrecan.Results In 12 weeks after cell-transplantation,standard T2-weighted image ratio(%ST2WI),mRNA expression levels of collagen II and aggrecan for DMEM group were signifcantly lower than that of DMEM group(P<0.05).While no significant differences between the NC and NPC groups were found.Histological analysis indicated that the degeneration score for the NC and NPC groups was signifcantly lower than the DMEM group(P<0.05).There was no significant difference between NC and NPC groups.Immunofluorescence staining of CollagenⅡand aggrecan indicated obviously positive in NC and NPC groups,and that of DMEM group gradually weakened with the change of time.Conclusion NPCs has a positive effect on restraining the discs degeneration.
作者
白亦光
陈巧玲
冯刚
刘康
肖东琴
林涛
罗栩伟
杨泽龙
BAI Yiguang;CHEN Qiaoling;FENG Gang;LIU Kang;XIAO Dongqin;LIN Tao;LUO Xuwei;YANG Zelong(Departments of Orthopaedics,The Second Clinical College of North Sichuan Medical College,Nanchong Central Hospital,Nanchong 637000,Sichuan,China;Departments of Oncology,The Second Clinical College of North Sichuan Medical College,Nanchong Central Hospital,Nanchong 637000,Sichuan,China;Institute of Tissue Engineering and Stem Cells,The Second Clinical College of North Sichuan Medical College,Nanchong Central Hospital,Nanchong 637000,Sichuan,China)
出处
《西部医学》
2020年第8期1122-1127,共6页
Medical Journal of West China
基金
四川省卫计委课题(16PJ202)
南充市校科技战略合作专项(18SXHZ0539)。
关键词
髓核细胞
椎间盘退行性变
细胞移植
早期
Nucleus pulposus cell
Intervertebral disc degeneration
Cell-transplantation
Early
