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- W2349002946 abstract "Objective To study whether the osteoclast is necessary for the anabolic effect of IGF-1 on the osteoblast in vitro.Methods Mouse MC3T3 osteoblast cells and RAW264.7 monocyte/macrophage cells were cultured in vitro.Receptor activator of nuclear factor kappa-B ligand(RANKL) was added to the RAW264.7 cell medium to induce the osteoclast differentiaion.Recombinant human insulin-like growth factor-1(rhIGF-1) 50 ng/ml were subjected to the treatment of osteoblasts and RAW264.7-derived osteoclasts,respectively.IGF-1 receptor activation was verified using Western blotting.The cultures of osteoblasts,osteoclasts,or the co-cultures of osteoblasts and osteoclasts were treated with 0,10,50,100 ng/ml of rhIGF-1,respectively.Osteoclast-conditioned medium with rhIGF-1 were collected after a 12-hour treatment.The conditioned medium was applied to the newly cultured osteoblasts for 12 hours.The concentrations of bone Gla protein(BGP) and RANKL in collected media were determined using ELISA.Bgp mRNA in the osteoblast was detected using real-time PCR.Results RAW264.7 cells could be successfully induced to mature multiple-nucleus and TRAP-positive osteoclasts by RANKL.Western blotting results confirmed that rhIGF-1 could effectively activate IGF-1 receptor phospharylation in both osteoblastsand osteoclasts.Results of ELISA and real-time PCR showed that there were no significant difference in either Bgp gene expression or BGP and RANKL concentrations in the medium between the IGF-1-additioned osteoblasts and the control cells.However,BGP and RANKL concentrations and the Bgp gene expression level were significantly elevated in both osteoblast cells cultured with osteoclast-conditioned medium and co-cultured cells.There was no significant difference between the osteoclast-conditioned culture group and the co-culture group.Conclusion Osteoclast is necessary for anabolic effect of IGF-1 on cultured osteoblast.The connection between the two cells may be completed through soluble cytokines." @default.
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- W2349002946 date "2010-01-01" @default.
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- W2349002946 title "The anabolic effect of IGF-1 on the osteoblast in the presence of the osteoclast" @default.
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