Pretreatments with hypertonic solution and cobalt chloride in bone marrow mesenchymal stem cell transplantation for treatment of degenerative disc disease

doi:10.3969/j.issn.2095-4344.2016.10.012

Abstract

Abstract:

BACKGROUND: Stem cell therapy has been used for prevention and treatment of degenerative disc disease. Considering the special microenvironment in the intervertebral disc, the survival rate and differentiation ability of transplanted cells are decreased, which may lead to the poor efficacy of stem cell therapy. How to improve the survival ability and therapeutic effect of the transplanted cells is the focus of stem cell therapy for degenerative disc disease.
OBJECTIVE: To investigate the effects of cobalt chloride combined with hypertonic solution pretreatment on bone marrow mesenchymal stem cells that will be transplanted for treatment of degenerative disc disease.
METHODS: (1) In vitro cell experiment: bone marrow mesenchymal stem cells were divided into three groups and subjected to normal culture medium (normal control group), 1% hypertonic mother solution (hypertonic group), 100 μmol/L cobalt chloride (hypoxia group), or 1% hypertonic mother solution plus
100 μmol/L cobalt chloride (combined group) for 1 week. Then, 2% hypertonic solution and 200 μmol/L cobalt chloride cobalt chloride were used to simulate the anaerobic and hypertonic environment intervenes in pretreated and untreated bone marrow mesenchymal stem cells for 24 hours. After that, RT-PCR was used to detect the expression of caspase-3 for apoptosis evaluation. (2) In vivo animal experiment: Sprague-Dawley rats were divided into model, cell transplantation and hypertonic plus hypoxic groups. Rat models of intervertebral disc degeneration were made in these three groups. After modeling, rats in these three groups were given no treatment, bone marrow mesenchymal stem cell transplantation or transplantation of bone marrow mesenchymal stem cells which were subjected to hypertonic and hypoxia pretreatments into the intervertebral disc. Two weeks later, immunohistochemistry and RT-PCR methods were used to detect cell distribution and related gene expression, respectively, thereby to evaluate the therapeutic effect of stem cells.
RESULTS AND CONCLUSION: (1) In vitro cell experiment: caspase-3 mRNA expression was significantly reduced in pretreated bone marrow mesenchymal stem cells compared with the untreated cells (P < 0.05). (2) In vivo animal experiment: compared with the control group, the caspase-3 and interleukin-1β in the intervertebral disc and a number of degenerative indexes were decreased in the cell transplantation. Compared with the cell transplantation group, these indicators had better outcomes in the hypertonic plus hypoxic group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells have therapeutic potential for degenerative disc disease, and have better adaptability and transplantation effects by hypertonic and hypoxia pretreatments.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Ye Han, Meng Zhen, Lin Jia-chen, Li Jia-wei, Zhang Yong-xing, Lin Nan-he, Zhao Qing-hua. Pretreatments with hypertonic solution and cobalt chloride in bone marrow mesenchymal stem cell transplantation for treatment of degenerative disc disease[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(10): 1452-1460.

Figures/Tables 5

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