Effect of Macrophage Colony-Stimulating Factor Overexpression on Osteoclast-related Cytokine Factors

doi:10.13701/j.cnki.kqyxyj.2019.11.019

Abstract

Abstract: Objective: To investigate the effect of macrophage colony-stimulating factor (M-CSF) overexpression in bone marrow mesenchymal stem cells (BMSCs) of SD rat on the osteoclast-associated cytokines. Methods: Overexpression plasmid vector to up-regulate M-CSF gene expression was constructed. The optimal transfection dose was determined using green fluorescent protein (EGFP) and the plasmid was transfected into BMSCs. The mRNA expression levels of osteoclast differentiation specific markers M-CSF, RANKL, TNF-α, IL-1, IL-6, and IL-24 were detected by reverse transcription polymerase chain reaction. Results: The positive rates of BMSCs surface markers CD29 and CD44 were 91.4% and 85.7% by flow cytometry and 2% by CD45. 5 μL/well LTX reagent was the best transfection efficiency. RT-PCR results showed that the expression of M-CSF in BMSCs of experimental group was significantly increased (P<0.001). The expression of RANKL, TNF-α, IL-1, and IL-24, the specific markers of osteoclast differentiation of BMSCs, increased significantly (P<0.001), while the expression of IL-6 decreased (P<0.001). There was a significant difference between the experimental group and the control group. Conclusion: Overexpression of M-CSF enhances the ability of BMSCs to differentiate into osteoclasts, and has an effect on the early, middle, and late stages of osteoclast differentiation. In addition, M-CSF high expression plasmid is a powerful gene therapy tool for bone regeneration by introducing foreign genes into BMSCs.

Key words: macrophage colony stimulating factor, osteoclast-related factors, osteoclastic signal pathway bone marrow mesenchymal stem cells, bone tissue engineering

WANG Jia, ZHANG Wenjing, HAN Xiangzhen, ZHOU Qiqi, HE Huiyu. Effect of Macrophage Colony-Stimulating Factor Overexpression on Osteoclast-related Cytokine Factors[J]. Journal of Oral Science Research, 2019, 35(11): 1089-1093.

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