Objective:To investigate the mechanism by which formononetin combined with platelet-rich plasma (PRP) enhances the proliferation and differentiation of osteoblasts.Methods:Rat osteoblasts ROS17/2.8 were cultured in vitro and treated with formononetin (10, 20, 40 μmol/L) combined with PRP. Cells were also intervened with G15 (a G protein-coupled estrogen receptor inhibitor) and Super-TDU [a Yes-related protein (YAP) inhibitor]. Cell proliferation viability was detected using the cell counting kit-8 (CCK-8) assay; alkaline phosphatase (ALP) activity was measured using an ALP assay kit; protein expression of G protein-coupled estrogen receptor (GPER) and p-YAP was detected by Western blot; and YAP subcellular distribution was analyzed by fluorescence assay.Results:Formononetin (20 μmol/L) synergistically enhanced the PRP-induced proliferation and ALP activity of osteoblasts (all P<0.05). Compared with the control group, formononetin significantly up-regulated GPER protein expression and down-regulated p-YAP protein expression (all P<0.01), with the most pronounced effects observed at 20 μmol/L. Formononetin (20 μmol/L) induced nuclear accumulation of YAP protein in osteoblasts. Pretreatment with G15 or Super-TDU reversed the synergistic effect of formononetin on PRP, and both the cell proliferation rate and ALP activity were lower than those in the PRP+ formononetin group (all P<0.01). Conclusions:Formononetin enhances the PRP-induced proliferation and differentiation of osteoblasts through the GPER/YAP signaling pathway.

Objective:To investigate the mechanism by which formononetin combined with platelet-rich plasma (PRP) enhances the proliferation and differentiation of osteoblasts.Methods:Rat osteoblasts ROS17/2.8 were cultured in vitro and treated with formononetin (10, 20, 40 μmol/L) combined with PRP. Cells were also intervened with G15 (a G protein-coupled estrogen receptor inhibitor) and Super-TDU [a Yes-related protein (YAP) inhibitor]. Cell proliferation viability was detected using the cell counting kit-8 (CCK-8) assay; alkaline phosphatase (ALP) activity was measured using an ALP assay kit; protein expression of G protein-coupled estrogen receptor (GPER) and p-YAP was detected by Western blot; and YAP subcellular distribution was analyzed by fluorescence assay.Results:Formononetin (20 μmol/L) synergistically enhanced the PRP-induced proliferation and ALP activity of osteoblasts (all P<0.05). Compared with the control group, formononetin significantly up-regulated GPER protein expression and down-regulated p-YAP protein expression (all P<0.01), with the most pronounced effects observed at 20 μmol/L. Formononetin (20 μmol/L) induced nuclear accumulation of YAP protein in osteoblasts. Pretreatment with G15 or Super-TDU reversed the synergistic effect of formononetin on PRP, and both the cell proliferation rate and ALP activity were lower than those in the PRP+ formononetin group (all P<0.01). Conclusions:Formononetin enhances the PRP-induced proliferation and differentiation of osteoblasts through the GPER/YAP signaling pathway.