髓核细胞和骨髓间充质干细胞对早期椎间盘退行性变干预作用对比  

Comparison of the effects of nucleus pulposus cells and bone marrow mesenchymal stem cells on the repair of early intervertebral disc degeneration

作　　者：白亦光[1] 陈巧玲[2] 刘康[3] 肖东琴[3] 罗栩伟[1] 林涛[1] 冯刚[3] Bai Yiguang;Chen Qiaoling;Liu Kang;Xiao Dongqin;Luo Xuwei;Lin Tao;Feng Gang(Department of Orhopaedics,Nanchong Central Hospital,The Second Clinical Medical College of North Sichuan Medical College,Nanchong 637000,China;Department of Oncology,Nanchong Central Hospital,The Second Clinical Medical College of North Sichuan Medical College,Nanchong 637000,China;Institute of Tissua Engineering and Stem Cells,Nanchong Central Hospital,The Second Clinical Medical College of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院第二临床医学院南充市中心医院骨科,南充637000 [2]川北医学院第二临床医学院南充市中心医院肿瘤科,南充637000 [3]川北医学院第二临床医学院南充市中心医院组织工程与干细胞研究所,南充637000

出　　处：《广西医科大学学报》2019年第12期1902-1908,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目（No.81171472、81071270、30872614）;四川省南充市校科技战略合作专项资助项目（No.18SXHZ0539）;四川省卫计委课题资助项目(No.16PJ202)

摘　　要：目的:比较髓核细胞(NPC)和骨髓间充质干细胞(BMSC)体外对兔椎间盘退行性变(IDD)修复效果。方法:体外分离、培养NPC、BMSC,并通过病理组织学染色和免疫组织化学染色对其进行鉴定。采用穿刺抽吸法建立兔IDD模型,造模时同期进行细胞移植治疗。将36只5月龄日本大耳白兔随机分为4组:正常对照组(NC组)、NPC移植组(NPC组)、BMSC移植组(BMSC组)、DMEM阴性对照组(DMEM组),每组9只。每只实验动物的L3-4、L4-5和L5-6作为实验干预椎间盘。术后4周、8周、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4周、8周、12周,NC组、NPC组和BMSC组的%ST2WI值及CollagenⅡ、aggrecan mRNA相对表达量均明显高于DMEM组(均P<0.05),NC组、NPC组和BMSC组组间比较均无明显差异(均P>0.05),DMEM组在术后4周、8周、12周%ST2WI值较NC组下降约50%、61%、70%;NC组、NPC组和BMSC组退变分数明显低于DMEM组(均P<0.05),NC组、NPC组和BMSC移植组组间比较均无明显差异(均P>0.05);NC组、NPC组和BMSC组的CollagenⅡ免疫组化结果显示强阳性,而DMEM组随着时间的推移阳性逐渐减弱。结论:BMSC和NPC移植在退变早期均能有效抑制椎间盘髓核组织的退变,BMSC有望成为NPC的良好替代者。Objective:To compare the effects of nucleus pulposus cell(NPC)and bone marrow mesenchymal stem cell(BMSC)on the repair of intervertebral disc degeneration in rabbits.Methods:NPC and BMSC were isolated and cultured.The intervertebral discs degeneration animal model was established by the conventional punctured method and at the same time NPC and BMSC were transplanted into the rabbit’s model of disc degeneration.Thirty-six 5-month-old Japanese white rabbits were randomly divided into 4 groups:normal control(NC)group,NPC transplantation group(NPC group),BMSC transplantation group(BMSC group)and DMEM negative control group(DMEM group).L3-4,L4-5 and L5-6 intervertebral discs were set as samples of experiment in rabbits.At week 4,8 and 12 after the transplantation,the repair effect of rabbits’degenerative intervertebral discs was evaluated by standard T2-weighted image ratio(%ST2WI),histopathology,degeneration score,immunochemical staining,and mRNA expression levels of Collagen II and aggrecan.Results:At week 4,8 and 12 after cell-transplantation,%ST2WI,mRNA levels of Collagen II and aggrecan in NC group,NPC group and BMSC group were significantly higher than those in DMEM group(P<0.05).No significant differences was found among NC,NPC and BMSC groups(P>0.05).ST2WI value in DMEM group decreased by about 50%,61%and 70%compared with NC group at 4,8 and 12 weeks after surgery.Histological analysis indicated that the degeneration score in NC,NPC and BMSC groups was lower than that in DMEM group(P<0.05).There was no significant difference among the NC,NPC and BMSC groups(P>0.05).Immunofluorescence staining of CollagenⅡand aggrecan indicated obviously positive in NC,NPC and BMSC groups,and that of DMEM group gradually weakened with time pronged.Conclusion:BMSC and NPC could restrain the intervertebral disc degeneration on early stage,and BMSC could be more effectively candidate of NPC.

关 键 词：髓核细胞 椎间盘退行性变 骨髓间充质干细胞 干细胞移植 

分 类 号：R394.2[医药卫生—医学遗传学]