Ion Channel Functional Protein Kinase TRPM7 Regulates Mg Ions To Promote The Osteoinduction Of Human Osteoblast

Objective In order to prove bone repair process simulation of biodegradable magnesium based implant in vitro,we examine the effects of magnesium ion on osteogenesis,migration,anti-alkaline stress through silencing of transient receptor cation channel 7(TRPM7),and explore the role of TRPM7 signaling pathway in magnesium ion-induced bone repair process.Method Magnesium ion concentration gradient(0.8,2,5,10,20,40 m M)was set up to treat osteoblast cell line h FOB1.19.Alizarin Red S staining,alkaline phosphatase staining and Transwell test were employed to determine the effect of magnesium ions on osteogenesis and migration of osteoblasts.By using si RNA interference of TRPM7 gene,alizarin red staining,alkaline phosphatase staining and Transwell test were employed again to determine the effect of osteogenesis via TRPM7 gene.5m M magnesium ion treatment with/without TRPM7 gene silencing group were set up,and the expression of MMP2,MMP9,VEGF,Runx2,ALP and BMP2 in each group was detected by PCR.A gradient p H values(p H=7,8,9,10)were established.Cytotoxicity test and flow cytometry cell death assay were used to observe the protective effect of magnesium ions on alkaline stress of osteoblasts.Result 1.The effective range of magnesium ions on promoting migration and osteogenesis was from 2m M to 10 m M.2.Magnesium ions increased the expression of MMP2,MMP9 and VEGF via TRPM7,thereby inducing osteoblasts to migrate from low magnesium concentration environment to high magnesium concentration environment.3.Magnesium ions increased the expression of Runx2 and ALP in osteoblasts via TRPM7,thereby enhancing osteogenesis of osteoblasts.4.The alkaline environment produced during the degradation of magnesium metal can induce cell death,but magnesium ions could rescue the cytotoxicity caused by alkaline stress via TRPM7.Conclusion Within the concentration range from 2mM to 10 mM,magnesium ions can enhance osteoblast osteogenesis,induce osteoblasts to migrate towards high magnesium concentration environment and protect osteoblast from alkaline stress via TRPM7.Therefore,this study provides a theoretical basis for molecular biology research and explores potential of biodegradable magnesium based implant in future clinical study.