The Study Of Preventive Effect Of ARC On BRONJ By Regulating BMSCs

Purpose:The impairment of bone marrow derived mesenchymal stem cells plays an important role during BRONJ development,while there is no effective treatment modility for BRONJ.In this study,we detect the proliferation and osteogenesis differentiation ability of BRONJ derived BMSCs and explor the possible molecular mechanism of the impairment ofthese cells.Further to explore the potential treatment effect of ARC overexpressed BMSCs on BRONJ.Material and methods:1.To isolate and culture BRONJ derived BMSCs,comparing the proliferation and osteogenesis differentiation ability with normal control group by MTT,ALP and ARS staining and RT-PCR.Ectopic new bone formation was conducted to evaluate the in vivo new bone formation ability of BRONJ derived BMSCs.Further experiment was used to detect the activation of TNF-α/NF-κB signal pathway.2.Establish ARC overexpressed BMSCs,culture the cells under hypoxia environment to micmic the in vivo hypoxia condition.MTT,flow cytometry analysis,ALP and ARS,immunoflurence staining were used to detect cell proliferation,apoptosis and osteogeneic differentiation respectively.Nude mice ectopic osteogenesis and calvarium defect rat mode were used to detect the in vivo new bone formation ability.RNA-seq was couducted to evaluate the differential gene expression.3.Coculture BRONJ derived BMSCs and ARC overexpressed BMSC.ALP,ARS and RT-PCR were used to evaluate the osteogenic differentiation ability.Infusion of ARC overexpressed BMSCs via the tail vein to evaluate the repairative effect on BRONJ mode.Results:1.The proliferation and osteogenic differentiation ability were decreased in BRONJ derived BMSCs.Activation of TNF-α/NF-κB signal pathway may involve in this process.2.ARC can promote BMSCs’ osteogenesis differentiation and inhibit its apoptosis caused by hypoxia.This process is mainly due to the inhibition of TNF-α/NF-κB signal pathway.3.ARC overexpressed BMSCs can promote osteogenesis differentiation of BRONJ derived BMSCs and are able to repair necrotic bone in BRONJ tissue.Conclusion:ARC gene regulates BMSCs by inhibiting TNF-α/NF-κB signal pathway to promote new bone formation in BRONJ tissue.