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使用定量T2成像评估骨髓间充质干细胞(BMSCs)植入对椎间盘再生的作用:一项兔实验研究

Evaluation of intervertebral disc regeneration with implantation of bone marrow mesenchymal stem cells (BMSCs) using quantitative T2 mapping: a study in rabbits.

作者信息

Cai Feng, Wu Xiao-Tao, Xie Xin-Hui, Wang Feng, Hong Xin, Zhuang Su-Yang, Zhu Lei, Rui Yun-Feng, Shi Rui

机构信息

Department of Orthopedics, Zhong Da Hospital, Southeast University, Nanjing, China.

出版信息

Int Orthop. 2015 Jan;39(1):149-59. doi: 10.1007/s00264-014-2481-0. Epub 2014 Aug 13.

Abstract

PURPOSE

The aim of the study was to investigate the curative effects of transplantation of bone marrow mesenchymal stem cells (BMSCs) on intervertebral disc regeneration and to investigate the feasibility of the quantitative T2 mapping method for evaluating repair of the nucleus pulposus after implantation of BMSCs.

METHODS

Forty-eight New Zealand white rabbits were used to establish the lumber disc degenerative model by stabbing the annulus fibrosus and then randomly divided into four groups, i.e. two weeks afterwards, BMSCs or phosphate-buffered saline (PBS) were transplanted into degenerative discs (BMSCs group and PBS group), while the operated rabbits without implantation of BMSCs or PBS served as the sham group and the rabbits without operation were used as the control group. At weeks two, six and ten after operation, the T2 values and disc height indices (DHI) were calculated by magnetic resonance imaging (MRI 3.0 T), and the gene expressions of type II collagen (COL2) and aggrecan (ACAN) in degenerative discs were evaluated by real-time reverse transcription polymerase chain reaction (RT-PCR). T2 values for the nucleus pulposus were correlated with ACAN or COL2 expression by regression analysis.

RESULTS

Cell clusters, disorganised fibres, interlamellar glycosaminoglycan (GAG) matrix and vascularisation were observed in lumber degenerative discs. BMSCs could be found to survive in intervertebral discs and differentiate into nucleus pulposus-like cells expressing COL2 and ACAN. The gene expression of COL2 and ACAN increased during ten weeks after transplantation as well as the T2 signal intensity and T2 value. The DHI in the BMSCs group decreased more slowly than that in PBS and sham groups. The T2 value correlated significantly with the gene expression of ACAN and COL2 in the nucleus pulposus.

CONCLUSIONS

Transplantation of BMSCs was able to promote the regeneration of degenerative discs. Quantitative and non-invasive T2 mapping could be used to evaluate the regeneration of the nucleus pulposus with good sensitivity.

摘要

目的

本研究旨在探讨骨髓间充质干细胞(BMSCs)移植对椎间盘再生的疗效,并研究定量T2映射方法评估BMSCs植入后髓核修复情况的可行性。

方法

采用48只新西兰白兔,通过穿刺纤维环建立腰椎间盘退变模型,然后随机分为四组,即两周后,将BMSCs或磷酸盐缓冲盐水(PBS)移植到退变椎间盘中(BMSCs组和PBS组),而未植入BMSCs或PBS的手术兔作为假手术组,未手术的兔作为对照组。在术后第2、6和10周,通过磁共振成像(3.0 T MRI)计算T2值和椎间盘高度指数(DHI),并通过实时逆转录聚合酶链反应(RT-PCR)评估退变椎间盘中II型胶原蛋白(COL2)和聚集蛋白聚糖(ACAN)的基因表达。通过回归分析将髓核的T2值与ACAN或COL2表达相关联。

结果

在腰椎退变椎间盘中观察到细胞簇、纤维紊乱、层间糖胺聚糖(GAG)基质和血管形成。发现BMSCs可在椎间盘中存活并分化为表达COL2和ACAN的髓核样细胞。移植后10周内,COL2和ACAN的基因表达以及T2信号强度和T2值均增加。BMSCs组的DHI下降比PBS组和假手术组更慢。髓核中的T2值与ACAN和COL2的基因表达显著相关。

结论

BMSCs移植能够促进退变椎间盘的再生。定量和非侵入性的T2映射可用于评估髓核的再生,具有良好的敏感性。

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