CORM-3 Promotes Osteogenic Differentiation of MC3T3-E1 Cells by Regulating the Polarization of Macrophages

doi:10.13701/j.cnki.kqyxyj.2022.03.017

Abstract

Abstract: Objective: To study the regulation of carbon monoxide releasing molecule-3 (CORM-3) on macrophage polarization, and to further explore the effects of CORM-3 mediated macrophage immune environment on proliferation and osteogenic differentiation of MC3T3-E1. Methods: RAW264.7 cells were cultured in vitro and the available CORM-3 concentration was screened. The expression of M1 and M2 related genes, such as iNOS, TGF-β, and IL-10, were detected by RT-qPCR. Immunofluorescence staining was used to detect the expression of Heme oxygenase-1 (HO-1) protein in each group. Flow cytometry was used to detect the proportion of M1 and M2 macrophages in each group after 24 hours. The supernatant after 24 hours was collected to prepare conditioned medium. CCK-8, alkaline phosphatase staining, alizarin red staining, and RT-qPCR were used to detect the proliferation and osteogenic activity of MC3T3-E1 cells. Results: RT-qPCR results showed that CORM-3 promoted the expression of TGF-β and IL-10, and decreased the expression of iNOS (P<0.05). Immunofluorescence results showed that CORM-3 increased the protein of HO-1 in RAW264.7 cells. In the environment where CORM-3 regulated macrophages, RT-qPCR results showed that CORM-3 increased the expression of OCN, Runx2, and Col-1 (P<0.05). The proliferation and osteogenic differentiation of MC3T3-E1 were enhanced. Conclusion: CORM-3 can increase the protein level of HO-1 by releasing CO, promote the polarization of macrophages RAW264.7 cells to the M2 phenotype, and enhance the osteogenic differentiation ability of MC3T3-E1 cells.

Key words: CORM-3, cell polarization, conditioned medium, osteogenic differentiation

WANG Yingying, DU Liuyi, LV Huixin, WANG Yuhui, REN Sicong, ZHOU Yanmin. CORM-3 Promotes Osteogenic Differentiation of MC3T3-E1 Cells by Regulating the Polarization of Macrophages[J]. Journal of Oral Science Research, 2022, 38(3): 280-285.

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