The Effect Study Of IL-10 Modified HAMSCs On The Factors Related To Fibrosis Of Wound Healing

Objective:Our previous studies have confirmed that the IL-10 modified human amniotic mesenchymal stem cells(hAMSCs_IL-10)is better than human amniotic mesenchymal stem cells?hAMSCs?in regulating wound inflammation,promoting blood vessel formation and accelerating the healing of the wound.This study investigated the effect and the mechanisms of hAMSCs _IL-10L-10 on the expression of factors in wound related to fibrosis in vivo and vitro,which provided a theoretical basis for clinical transformation.Methods:1.The effect of hAMSCs_IL-10L-10 on the expression of fibrosis-related factors in fibroblasts in vitroFirst,hAMSCs_IL-10L-10 were constructed and the IL-10 in the supernatant was determined.To investigate the effect of hAMSCs and hAMSCs_IL-10L-10 on the migration of human dermal fibroblasts,we collected their conditioned medium and act on fibroblasts to observe cell migration.To study the mechanism,we co-cultured human dermal fibroblasts with hAMSCs or hAMSCs_IL-10L-10 in Transwell and the expression of collagen I,collagen III,DCN,alpha-SMA and HAS2 in fibroblasts were detected by RT-PCR and Western blot.2.The effect of hAMSCs_IL-10L-10 on the expression of fibrosis-related factors in wound healing fibrosis in miceWild type C57BL/6 mice were used to establish the full-thickness skin defecting model which were divided into three groups:control,hAMSCs and hAMSCs_IL-10L-10 groups.The wound healing condition and histological changes of mice were observed after injections of PBS,hAMSCs and hAMSCs_IL-10L-10 into the wound locally.The expression of collagen type I,collagen type III,DCN,alpha SMA and HAS2 were detected by ELISA in mice wound extracellular matrix.Results:1.The effect of hAMSCs_IL-10L-10 on the expression of fibrosis-related factors in fibroblasts in vitroThe level of IL-10 in the medium of hAMSCs_IL-10L-10 was higher than hAMSCs.Both conditioned medium of hAMSCs and hAMSCs_IL-10L-10 could promote the migration of human fibroblasts,and the effect of hAMSCs_IL-10L-10 was better.The hAMSCs_IL-10L-10 group could down-regulate the expression of collagen I and alpha-SMA and up-regulate the expression of collagen III,DCN and HAS2in fibroblasts,and hAMSCs_IL-10L-10 had a stronger regulatory effect to hAMSCs.2.The effect of hAMSCs_IL-10L-10 on the expression of fibrosis-related factors in wound healing fibrosis in miceThe infiltration of inflammatory cells in hAMSCs group and hAMSCs_IL-10L-10 group were less than that of the control group.Compared with hAMSC,the hAMSCs_IL-10L-10 group had less inflammatory cell infiltration and the wound healing was faster.On both day 7 and 14after model establishment,the expression of type I collagen and?-SMA were much more decreased and the expression of type III collagen,DCN and HAS2 were much more increased in hAMSCs_IL-10L-10 group compared with hAMSCs group.It was revealed that there was still GFP positive cells in wound locally on 14 day after transplantation in hAMSCs_IL-10L-10 group in vivo.Conclusion:1.Compared with hAMSCs,hAMSCs_IL-10L-10 promote the migration of fibroblasts more effectively in vitro,up-regulate the expression of anti-fibrotic factors DCN,HAS2,and III collagen,and down-regulate the expression of profibrotic factors type I collagen and?-SMA.2.Local transplantation of hAMSCs_IL-10L-10 can up-regulate the expression of anti-fibrotic factors such as DCN,type III collagen and HAS2 in mouse skin wounds,down-regulate the expression of profibrotic factors such as type I collagen and?-SMA.In addition,hAMSCs_IL-10L-10 can promote wound healing of C57BL/6 mice,reduce the degree of fibrosis of scars,and improve the quality of healing.