Promotion of Osteoblasts Differentiation in Sinusoidal Electromagnetic Fields Involves BMP-Smad1/5/8 Signal Pathway
XIE Yan-Fang 1), SHI Wen-Gui 2), ZHOU Jian1),GAO Yu-Hai 1), WANG Ming-Gang 3), CHEN Ke-Ming1)*
1) Institute of Orthopaedics, Lanzhou General Hospital,Lanzhou Command of CPLA, Lanzhou 730050, China;
2)Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730050,China;
3) School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,China
Abstract：Sinusoidal electromagnetic fields (SEMFs) have been considered as a stimulator for rat osteoblasts (ROBs) differentiation. However, the mechanism of the SEMFs-induced osteogenesis is still unknown. In this study, we examined the role of the BMP-Smad signaling pathway in SEMF-induced differentiation of primary ROBs. Cells were obtained by sequential enzyme digestion from the segregated skull of neonatal SD rats. The sub-cultured ROBs were treated in 50Hz SEMFs at 1.8 mTs for 0, 5, 15, 30, 60 and 120 min before measuring for BMP-2, total or phospho-Smad1/5/8 proteins by Western blotting or immunofluorescence staining. Noggin 10 nmol/L was used to block the BMP2-Smad1/5/8 signaling. Intracellular alkaline phosphatase (ALP) activity was detected at 3 and 6 days after cells were treated by SEMFs for 120 min/day. The mRNA and protein expression levels of RUNX-2 and type I collagen were measured at day 2 by real-time PCR and Western blotting. The results showed that SEMFs promoted the expression of BMP-2 and rapidly induced the phosphorylation and nuclear translocation of Smad1/5/8 in ROBs. The treatments also significantly promoted ALP activity and the expression of RUNX-2 and type I collagen. The SEMFs-induced osteoblast gene expression could be blocked by noggin. Collectively, we demonstrated that SEMFs promoted osteoblast differentiation by modulating the BMP2-Smad1/5/8 signaling.
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