The Preliminary Study On The Mechanism Of Gingiva-derived Mesenchymal Stem Cells In The Therapy Of Mice Radioactive Stomatitis

Purpose:The incidence of radioactive stomatitis in head or neck cancer patients after radioactive treatment is high,which severely affect the effect of therapeutic and quality of life.As a new kind of precursor mesenchymal stem cells,GMSCs(gingiva-derived mesenchymal stem cells)has similar characteristics and functions with other mesenchymal stem cells such as adipose mesenchymal stem cells and bone-marrow mesenchymal stem cells,as well as the incomparable advantages.In this experiment,GMSCs were applied into therapy of mice radioactive somatitis aiming to explore the mechanism of this process,and providing new evidence and thinking in the prevention and treatment of the radioactive stomatitis.Methods:1.Isolate and culture of GMSCs of GFP+ transgenic C57/BL mice by ensymoly method2.Select 24 C57/BL mice of 6-8 weeks and randomly divided into 3 groups as follows:(1)GMSC control group:no radiation,inject of 106 GMSCs by tail veins(2)Radiation treatment group:use electron linear accelerator to build radiation mice model,in the dose of 25Gy each time,inject of 106 GMSCs by tail veins 3 days after radiation(3)Radiation control group:use electron linear accelerator to build radiation mice model,in the dose of 25Gy each time,inject of cell-free DMEM medium by tail veins 3 days after radiation3.Establish animal models of C57/BL mice of radioactive stomatitis:use electron linear accelerator to establish radiation mice model,with the dose of 25Gy each time,and then observe pathological change under optical microscope.4.Observe GMSCs homing in the lips and tongue tissue in mice,analyse the changement of cytokine in lips and tongue tissue in mice(1)GMSC control group:Cut perioral tissues and of mice 3 days,7 days and 10 days after injection of the GMSCs,detect the GFP+-GMSCs homing by immunohistochemical;(2)Radiation treatment group:Cut perioral tissues and of mice 3 days,7 days and 10 days after injection of the GMSCs,detect the GFP+-GMSCs homing by immunohistochemical;meanwhile detect the expression of IL-6,IL-10,SDF-1 in the perioral tissues by enzyme-linked immuno sorben assay(ELISA).(3)Radiation control group:detect the expression of IL-6,IL-10,SDF-1 in the perioral tissues by enzyme-linked immuno sorben assay(ELISA)after injection of DMEM Results:1?Separation and culture of GMSCs of C57/BL mice;2?The immunohistochemical results of GMSCs homing in the lip and tongue of radioactive stomatitis mice model:There is no obvious GFP+GMSCs cell homing in GMSC control group;the GFP+GMSCs cell homing can be observed in epithelium of lip and tongue,with the largest number at 3 days after injection,and diminishing at 7 days,10 days after injection.3?The expression of IL-6,IL-10,SDF-1 in the perioral tissues of mice:3 days?7days?10days after injection,the expression of IL-6 in Radiation treatment group is lower than Radiation control group(p<0.05),the expression of IL-10 in Radiation treatment group is higher than Radiation control group(p<0.05),the expression of SDF-1 in Radiation treatment group is higher than Radiation control group(p<0.05);Conclusion:After systematically application GMSCs to radioactive stomatitis in mice,the expression of SDF-1 increase in local tissues,thus promotes the GFP + GMSCs homing in cortex on lips and tongue,at the same time,expression of IL-6 decrease and IL-10 increas,which is one of the mechanism for GMSCs treatment of radioactive stomatitis in mice.