The impact of macrophage polarization derived from THP-1 cells on the proliferation and differentiation of dental pulp stem cells
Objective:To evaluate the potential impact of macrophages differentiated from the human monocytic leu-kemia cells line THP-1 under different polarization states on the growth and differentiation potential of human dental pulp stem cells(hDPSC).Methods:THP-1 cells were induced to differentiate into M1 and M2 macrophages under specific condi-tions,and the differentiation outcomes were verified using immunofluorescence and flow cytometry techniques.The expression levels of macrophage polarization marker genes were assessed through qRT-PCR analysis.In the experiment,hDPSC were di-vided into three groups:the control group,the M1 macrophage culture supernatant treatment group,and the M2 macrophage culture supernatant treatment group.The proliferation of hDPSC was evaluated using the CCK-8 assay,the mineralization ca-pacity was detected by alizarin red and alkaline phosphatase(ALP)staining,and the expression of proteins related to odonto-genic differentiation was examined using Western blot technology.Additionally,potential molecular mechanisms were explored through cellular transcriptome analysis.Results:THP-1 cells were induced to differentiate into M1 and M2 macrophages un-der different stimuli.The expression of pro-inflammatory cytokines such as tumor necrosis factor-alpha(TNF-α)and interleu-kin-6(IL-6)in M1 macrophages was significantly increased(P<0.05),while the expression of anti-inflammatory cytokines such as transforming growth factor-beta(TGF-β)and IL-10 in M2 macrophages was significantly increased(P<0.05).hDPSC treated with M2 macrophage culture supernatant showed a higher proliferation rate compared to the control and M1 treatment groups(P<0.05),and alizarin red staining revealed more calcium nodule formation,with a significant increase in the expres-sion of DSPP and DMP-1 proteins(P<0.05).Transcriptome analysis revealed that compared to the control group,there were 326 upregulated genes and 347 downregulated genes in the M2 macrophage treatment group,which are mainly involved in the PI3K-Akt and MAPK signaling pathways.Conclusion:M2 macrophages derived from THP-1 can enhance the proliferation of hDPSC,release anti-inflammatory cytokines,and enhance the odontoblast differentiation capacity of hDPSC,with the mecha-nism of action possibly being through the PI3K-Akt and MAPK signaling pathways.
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