TGFβ1干预下体内兔骨髓间充质干细胞对退变椎间盘治疗的实验研究
骨髓间充质干细胞;,椎间盘;,转化生长因子β1(TGFβ1);,髓核细胞,,骨髓间充质干细胞;,椎间盘;,转化生长因子β1(TGFβ1);,髓核细胞,TGFβ1干预下体内兔骨髓间充质干
摘 要:[目的]评价在TGFβ1干预下的兔间充质干细胞移植到兔退变椎间盘后是否可诱导向髓核细胞分化并增加蛋白多糖和Ⅱ型胶原的含量。[方法]采用体外细胞培养技术,将兔骨髓间充质干细胞自骨髓血中分离、纯化和培养,成功传代、纯化后并在TGFβ1干预下植入兔退变椎间盘的模型中。分别于2、4、6、8周用间苯三酚分光光度法测定蛋白多糖含量的变化;免疫组化法测定Ⅱ型胶原的含量变化。所得数据经SPSS 115统计学软件进行统计学处理。[结果]原代培养及传代培养显示兔骨髓间充质干细胞具有活跃的增殖倍增能力,并且8周内蛋白多糖和Ⅱ型胶原的含量的恢复实验组对比模型组增高明显,统计学显示差异有显著性意义。[结论]将TGFβ1干预下的兔间充质干细胞移植到退变椎间盘后可以在一定时间内增加蛋白多糖和Ⅱ型胶原的含量,从而延缓椎间盘退变。关键词:骨髓间充质干细胞; 椎间盘; 转化生长因子β1(TGFβ1); 髓核细胞
Intervertebral disc cell therapy for regeneration:mesenchymal stem cell cultivation in vitro and intervention in TGFβ1 implantation in rabbit intervertebral discs∥ZHAO Ziru,WU Xiaotao,QI Yabin,et alDepartment of Orthopaedics,Zhongda Hospital of Southeast University,Nanjing 210009,China
Abstract:[Objective]To evaluate whether transplanted marrow mesenchymal stem cells interfered in TGFβ1 can differentiate to nucleus pulposus cells and increase the amount of proteoglycan and collagenase Ⅱ content in intervertebral discs[Method]We used an in vivo model to investigate the feasibility of marrow mesenchymal stem cells that cultured in vitro and interfered in TGFβ1 delivery,retention,and survival in the degeneratived disc spaceIn 2,4,6,8 weeks we used immunohistochemical staining to determine the change of collagenase Ⅱ content;spectrophotometry to determine the change of amount of proteoglycan with Phlorglucinol;the experiment date were analyzed by SPSS 115 soft ware[Result]We found MSCs could maintain viability and proliferate within the rabbit inter vertebral discThe amount of proteoglycan and collagen Type Ⅱ content of the intervertebral in matrix synthesis in the experiment group was increased in 8 weeksWe found no changes in the modle group[Conclusion]Our data suggest that transplanted marrow mesenchymal stem cells in vivo can survive and increase proteoglycan and collagen Type Ⅱ amount interfered in TGFβ1 in some periods,which support its potential use as a treatment of intervertebral disc degeneration ......
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