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研究设计:骨形态发生蛋白2联合富血小板血浆治疗椎间盘退变的体外和体内评估

Study design: in vitro and in vivo assessment of bone morphogenic protein 2 combined with platelet-rich plasma on treatment of disc degeneration.

作者信息

Hou Yang, Shi Guodong, Shi Jiangang, Xu Guohua, Guo Yongfei, Xu Peng

机构信息

Orthopaedic Department, Changzheng Hospital, Shanghai, China.

出版信息

Int Orthop. 2016 Jun;40(6):1143-55. doi: 10.1007/s00264-015-2840-5. Epub 2015 Jul 14.

Abstract

OBJECTIVE

Our aim was to investigate the biological effects of bone morphogenic protein 2 (BMP2) on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into chondrocyte-like cells in platelet-rich plasma (PRP) gel in vitro. In addition, the effectiveness of BMP2-transduced BMSCs in combined with PRP gel to repair the degenerated intervertebral disc in a rabbit model was also evaluated. Previous studies have shown that tissue engineering provides many promising advantages to treating disc degeneration and may reverse the pathological process of disc degeneration.

METHODS

The expressions of types I, II and X collagen, aggrecan and Sox9 of the BMP2-transduced BMSCs in monolayer or PRP gel were examined by reverse transcriptase polymerase chain reaction (RT-PCR). Sixty New Zealand white rabbits were divided into five groups: 12 normal controls; 12 puncture operated with only disc degeneration being induced; 12 PRP transplantation animals; 12 BMSC and PRP-transplantation animals; 12 BMP2-transduced BMSCs and PRP-transplantation animals. The effect of BMP2-transduced BMSCs on degenerated discs were evaluated by magnetic resonance image (MRI) scan, histology, immunohistochemistry and Western blot analysis.

RESULTS

BMP2 could facilitate chondrogenic differentiation of BMSCs in monolayer or PRP gel. The discs treated with BMP2-transduced BMSCs exhibited relatively well-preserved nucleus pulposus (NP) structure. Significantly higher T2-weighted signal intensity and a greater amount of extracellular matrix were observed in the BMP2-transduced BMSC group compared with other groups. In addition, the presences of BMP2-transduced BMSCs were identified at week 12 postoperatively in vivo.

CONCLUSIONS

BMP2-transduced BMSCs can maintain the chondrocyte-like phenotype in PRP gel in vitro, and the combined use of these two agents can significantly promote repair of the degenerated discs in vivo.

摘要

目的

我们的目的是研究骨形态发生蛋白2(BMP2)对体外富血小板血浆(PRP)凝胶中骨髓间充质干细胞(BMSCs)向软骨样细胞分化的生物学效应。此外,还评估了经BMP2转导的BMSCs联合PRP凝胶修复兔模型中退变椎间盘的有效性。先前的研究表明,组织工程为治疗椎间盘退变提供了许多有前景的优势,并且可能逆转椎间盘退变的病理过程。

方法

通过逆转录聚合酶链反应(RT-PCR)检测经BMP2转导的BMSCs在单层或PRP凝胶中I型、II型和X型胶原蛋白、聚集蛋白聚糖和Sox9的表达。60只新西兰白兔分为五组:12只正常对照组;12只仅诱导椎间盘退变的穿刺手术组;12只PRP移植组动物;12只BMSC和PRP移植组动物;12只经BMP2转导的BMSCs和PRP移植组动物。通过磁共振成像(MRI)扫描、组织学、免疫组织化学和蛋白质印迹分析评估经BMP2转导的BMSCs对退变椎间盘的作用。

结果

BMP2可促进单层或PRP凝胶中BMSCs的软骨形成分化。经BMP2转导的BMSCs治疗的椎间盘髓核(NP)结构保存相对良好。与其他组相比,经BMP2转导的BMSC组观察到显著更高的T2加权信号强度和更多的细胞外基质。此外,术后12周在体内鉴定出经BMP2转导的BMSCs的存在。

结论

经BMP2转导的BMSCs在体外PRP凝胶中可维持软骨样表型,并且这两种制剂的联合使用可显著促进体内退变椎间盘的修复。

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