AGS3 Is Involved In TNF-α Medicated Osteogenic Differentiation Of Human Dental Pulp Stem Cells

Objective To observe the effect of Activator of G-protein signaling 3(AGS3)in TNF-α medicated osteogenic differentiation of human dental pulp stem cells.Methods We isolated and cultured DPSCs.The multipotency of DPSCs was identified by chondrogenic and adipogenic differentiation of DPSCs was demonstrated by positive staining with toluidine blue and Oil Red O.Human DPSCs were cultured in osteogenic differentiation and supplied with TNF-α(0.01,100ng/mL)for 14 days.Alizarin red S staining and the expression of alkaline phosphatase(ALP),bone morphogenetic protein 2(BMP2),runt-related transcription factor 2(Runx2)were dectect by Western blot.The expression of AGS3 were dectected by Western blot and RT-PCR.Western blot analyzed p-CREB expression and PKA activity.The negative role of AGS3 in osteogenic differentiation was further confirmed by knocking down and over expression of AGS3.The expression of p-CREB were detected by Western blot.Results DPSCs exhibited a fibroblast-like morphology.DPSCs could differentiate into chondrogenic cells and adipogenic cells.In this study,we found that TNF-α(0.01ng/mL)promoted the osteogenic differentiation of DPSCs while 100ng/mL decreased thedifferentiation.Increased expression of AGS3 inhibited p-CREB expression and PKA activity exposed to TNF-α(100ng/mL).Conclusions Our studies confirmed that high level TNF-α(100ng/mL)inhibited osteogenic differentiation of DPSCs.AGS3 is involved in TNF-α treated osteogenic differentiation of DPSCs via c AMP/PKA/CREB signaling pathway.Our findings may provide clinical implications for osteoporosis.