The Effects Of Interferon-? On The Proliferation And Osteogenic Differentiation Of SHED In Vitro

Objective:Oral and maxillofacial trauma,tumor,congenital malformation and other reasons can cause large defects in the maxillofacial bone tissue,that affect the patient's chewing function,pronunciation and appearance seriously.Stem cell-based tissue regenerative medicine is a new direction for the treatment of maxillofacial bone tissue defects.In recent years,studies have found that the host immune microenvironment,ie,immune cells and their secreted inflammatory cytokines play a crucial role in tissue regeneration,which largely regulates the biological properties of stem cells,participates in and determines the efficacy of organization regeneration.Previous studies have found that the pro-inflammatory cytokine interferon-gamma(IFN-?)may be associated with osteogenic ability of stem cells from exfoliated deciduous teeth(SHED),but the specific regulation is unknown.In this experiment,we will investigate the effects of IFN-? on the proliferation and osteogenic differentiation of SHED cells by in vitro cell experiments,and provide a basis for clarifying the role of immune microenvironment in tissue regeneration,and seek new ways to improve the efficacy of stem cell-based bone tissue regeneration.Methods : Human SHED were primary isolated with enzymatic digestion.Flow cytometry was used to evaluate surface marker expression of SHED,such as CD34,CD45,CD73,CD90,and CD105.Alizarin red S,oil red O staining and Immunohistochemistry were used to evaluate the multi-lineage differentiation potential of SHED.SHED was treated with different concentrations of IFN-? 0,10,50,100,200 ng/m L).The effect of IFN-? on the proliferation of SHED cells was detected by CCK8 method.The ratio of SHED apoptosis was detected by flow cytometry and toluidine blue staining.To identify the osteogenic differention of SHED pretreated by different concentrations of IFN-?(0,10,50,100,200 ng/m L),the protein expressions of Runx2 and ALP were examined by Western Blot.The capacity of mineralized nodule formation was observed by Alizarin red S staining.The data were analyzed by SPSS 18.0 software.Results:IFN-? could inhibit the proliferation of SHED in a concentration-dependent manner.The higher the concentration of IFN-? was,the more obvious the effect of inhibiting the proliferation of SHED.High concentrations of IFN-?(100,200 ng/m L)could induce apoptosis of SHEDand inhibited the osteogenesis of SHED.Conclusions : IFN-? induces apoptosis of SHED,inhibits cell proliferation and osteogenic differentiation,and reduces the effect of SHED-based bone regeneration.Regulating the recipient immune microenvironment is a new important target for improving the efficacy of stem cell-based bone tissue regeneration.