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[大鼠同种异体骨髓间充质干细胞移植治疗脊髓损伤的时间能量分析]

[Analysis of chronergy for treatment of spinal cord injury with the allogeneic bone mesenchymal stem cells (BMscs) transplantation in rats].

作者信息

Duan Da-peng, Su Quan, Hu Wei, You Wu-Lin, Dang Xiao-Qian, Wang Kun-Zheng

机构信息

Department of Orthopaedics ,the Second Affiliated Hospital of Xi' aon Jiaootong University, Xi 'an 710004, Shaanxi, China.

出版信息

Zhongguo Gu Shang. 2013 Oct;26(10):845-9.

Abstract

OBJECTIVE

To observe the change of behavior, pathological change of the spinal cord,and expression of brain-derived neurotrophic factor (BDNF) and brain-derived neurotrophic factor (NGF) on rats with spinal cord injury in order to explore the optimal time of BMSCs transplantation.

METHODS

Eighty health SD rats were randomly divided into 8 groups (group A,B,C,D,E,F,G,H), 10 rats in each group. According to the modified Allen method,the rat model of spinal cord injury was built. Group A as non-injured group only exposed the spinal cord but not result in blast injury. BMSCs of vitro culture were respectively infunded the region of spinal cord injury in group C, D, E, F, G, H (as transplantation groups) at the 0 h, 6 h, 24 h,3 d,5 d,7 d after model made. Group B as single model group was infunded the equal cell culture fluid. BBB score was used to evaluate the function of spinal cord at the 1st,2nd and 4th weeks after injury. The morphological changes of the tissue of spinal cord injury were observed by HE stain and the expression of BDNF and NGF were detected by Elisa method at the 4th weeks after BMSCs transplantation.

RESULTS

In non-injured group,BBB score was highest than that of other 7 groups at the 1st, 2nd and 4th weeks after injury (P<0.01). There was no significant difference in BBB score between single model group and transplantation groups at the 1st week after BMSCs transplantation (P>0.05). BBB score in transplantation groups were higher than that of single model group at the 2nd and 4th weeks after BMSCs transplantation (P<0.05). At the 2nd week after injury, BBB score from high to low was group F,E,G,D,H,C,but there was no significant difference among the groups (P>0.05). At the 4th week after injury,there was significant differences in BBB score between group F and other transplantation groups (group C,D,E,G,H)(P<O.05),but there was no significant difference among 5 groups (P>0.05). The levels of BDNF and NGF in group F was highest at the 4th week after BMSCs transplantation (P<0.05). In non-injured group,HE staining showed the struction of spinal cord was clear and complete,no neutrophil was found. But in other 7 groups,HE staining showed obviously oedema at local tissue,juncture of gray and white matter was undefined,and different gliocyte proliferation and inflammatory cell infiltrate cound be found.

CONCLUSION

Allogeneic BMSCs transplantation is effective to stimulate the recovery of spinal cord function in rats with spinal cord injury,and the optimal time of BMSCs transplantation maybe at the 3 d after injury.

摘要

目的

观察脊髓损伤大鼠的行为变化、脊髓病理改变以及脑源性神经营养因子(BDNF)和神经生长因子(NGF)的表达,以探讨骨髓间充质干细胞(BMSCs)移植的最佳时机。

方法

将80只健康SD大鼠随机分为8组(A、B、C、D、E、F、G、H组),每组10只。采用改良Allen法建立大鼠脊髓损伤模型。A组作为未损伤组,仅暴露脊髓但不造成冲击损伤。C、D、E、F、G、H组(作为移植组)在造模后0小时、6小时、24小时、3天、5天、7天分别将体外培养的BMSCs注入脊髓损伤区域。B组作为单纯模型组,注入等量细胞培养液。在损伤后第1、2、4周采用BBB评分评估脊髓功能。在BMSCs移植后第4周,通过HE染色观察脊髓损伤组织的形态学变化,采用Elisa法检测BDNF和NGF的表达。

结果

在损伤后第1、2、4周,未损伤组的BBB评分高于其他7组(P<0.01)。在BMSCs移植后第1周,单纯模型组与移植组的BBB评分无显著差异(P>0.05)。在BMSCs移植后第2、4周,移植组的BBB评分高于单纯模型组(P<0.05)。在损伤后第2周,BBB评分从高到低依次为F组、E组、G组、D组、H组、C组,但各组间无显著差异(P>0.05)。在损伤后第4周,F组与其他移植组(C、D、E、G、H组)的BBB评分有显著差异(P<0.05),但这5组间无显著差异(P>0.05)。在BMSCs移植后第4周,F组的BDNF和NGF水平最高(P<0.05)。未损伤组HE染色显示脊髓结构清晰完整,未见中性粒细胞。但其他7组HE染色显示局部组织明显水肿,灰质和白质交界处不清,可见不同程度的胶质细胞增生和炎性细胞浸润。

结论

异体BMSCs移植对促进脊髓损伤大鼠脊髓功能恢复有效,BMSCs移植的最佳时机可能为损伤后3天。

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