| Background:Mesenchymal stem cells(MSCs)possess a remarkable capacity for self-renewal and multi-directional differentiation,thus offering a wide range of possibilities in the realm of regenerative medicine.The purpose and operation of mesenchymal stem cells,such as bone marrow mesenchymal stem cells,adipose mesenchymal stem cells and umbilical cord mesenchymal stem cells,has been extensively researched and has been progressively implemented in clinical practice.Compared with adult mesenchymal stem cells,fetal mesenchymal stem cells not only have higher proliferation ability and multi-directional differentiation potential,but also have lower immunogenicity.These advantages make fetal mesenchymal stem cells attract wide attention.Fetal skin-derived stem cells(FSDC),extracted from fetal skin tissue in the second trimester of pregnancy,are the main cells mediating fetal skin scarless healing and have potential advantages in the field of skin tissue repair and regeneration.In previous experiments,we detected that it expressed mesenchymal stem cell-specific surface markers,suggesting that it may be a mesenchymal stem cell.The FDSCs,in comparison to other stem cells,possess a more potent capacity for proliferation and migration,and may possess exclusive benefits.At present,there are few studies on the application of FDSCs in wound treatment,and the effect and mechanism of FDSCs in wound treatment are still unclear,which needs further investigation.Objective:The aim of this research is to explore the influence of FDSCs on the healing of skin wounds,to determine if FDSCs are more likely to act as seed cells for wound healing cell therapy when compared to ADSCs,and to investigate the potential mechanism of their influence on skin wound healing.Methods:1.FDSCs and ADSCs were co-cultured with HSF,and the effects of the two kinds of cells on the proliferation of HSF were compared by CCK-8 method.The effects of the two kinds of cells on the migration of HSF were compared by scratch test.The effects of the two kinds of cells on the collagen secretion of HSF were detected by immunofluorescence.2.FDSCs and ADSCs were co-cultured with HUVECs,respectively.The effects of the two kinds of stem cells on the proliferation of HUVECs were compared by CCK-8method.The effects of the two kinds of stem cells on the formation of HUVECs were compared by cell chamber formation test.3.Establishing a model of full-thickness skin defect wound on the backs of nude mice.The cells were injected into four points around the wound in the experimental group immediately after operation.The healing of the skin wound was observed on the7 th,14th and 21 st day after operation,and photographed and recorded.On the 21 st,HE and Masson staining were performed on the wound tissue for observation and comparison.4.The wound tissues on different day were taken for protein extraction.Elisa method detection of inflammatory factors in wound tissues of nude mice.5.WB was used to detect the expression of collagen type I and III.6.The expression of VEGF in the wound tissue of nude mice was identified by Elisa,while WB was utilized to identify CD31 on the 21 st day of the wound.Results:1.After co-culture of FDSCs,ADSCs and HSF,both stem cells can promote the proliferation and migration of HSF.The relative viability and migration rate of HSF in co-culture of FDSCs and HSF were higher than that in co-culture of ADSCs.2.Both stem cells could promote the ability of HSF to secrete collagen type Ⅰand Ⅲ.When FDSCs were co-cultured with HSF,HSF secreted more type I and type III collagen,and effectively reduced the ratio of type I/type III.3.Both stem cells could enhance the proliferation and tube formation of HUVECs.Moreover,FDSCs induced HUVECs to proliferate more strongly,and the number of tube branches and tube area increased significantly.4.In the nude mouse wound model,it was observed that the wound in the FDSCs intervention group had healed more than half on the 7th day after operation and tended to heal on the 14 th day,while the ADSCs intervention group had some defect on the14 th day,and the Control group had a large gap on the 14 th day and just healed on the21 st day.At 21 days,the scar from FDSCs had become almost smooth,yet the residual scar tissue in ADSCS was visible,and the scar tissue was conspicuous in the control group.Statistical analysis revealed that the wound healing rate of the FSDSCs intervention group and ADSCs intervention group was faster than that of the control group,while the FDSCs intervention group’s wound healing rate was faster than that of the ADSCs intervention group.5.According to the HE staining results,FDSCs showed more obvious reepithelialization and less inflammatory cell infiltration than ADSCs.The results of Masson staining revealed that the pathological alterationsof FDSCs and ADSCs intervention group were relatively orderly compared with the control group,and the improvement of FDSCs group was more significant than that of ADSCs group.6.The expression of TNF-α in the FDSCs intervention group was lower than that in the ADSCs intervention group and the control group at day 3,and lower than that in the control group at days 7 and 21.The expression of IL-6 in the FDSCs intervention group was significantly lower than that in the ADSCs intervention group and the control group at days 14 and 21,and lower than that in the control group at day 3.The expression of IL-13 in the trabeculae of the FDSCs intervention group was significantly lower than that of the ADSCs intervention group and the control group on day 14 and on day 3.The expression of IL-10 in the trabeculae of the FDSCs intervention group was significantly lower than that of the ADSCs intervention group and the control group on day 21 and on days 3,7 and 14.Statistical analysis revealed that the FDSCs intervention group was more successful in decreasing inflammatory factors and augmenting anti-inflammatory factors than the ADSCs intervention group,as evidenced by their significantly lower expression of IL-10 in the trabeculae on days 21 and 7 and 14 respectively.7.WB results showed that both stem cells could promote the production of collagenⅠ and collagen Ⅲ,but FDSCs group had more obvious ability to promote collagen synthesis and could better improve the proportion of collagen Ⅲ in wound.8.FDSCs can promote angiogenesis.Gross observation showed that DSCs intervention group compared with ADSCs intervention group and control group(injection of PBS)significantly increased the number of blood vessels,more branches.The WB results showed that the expression of CD31 in FDSCs group was higher than that in ADSCs intervention group and control group.The expression of VEGF in wound surface was detected by Elisa.Conclusions:1.Compared with ADSCs,FDSCs could enhance the proliferation and migration of HSF.2.Compared with ADSCs,FDSCs can promote HSF to secrete collagen more,and can increase the proportion of type III collagen secretion.3.Compared with ADSCs,FDSCS could enhance the proliferation and angiogenesis of HUEVCs.4.Compared with ADSCs,FDSCS can promote the healing of skin wounds on the back of mice.5.The role of FDSCs in promoting wound may be related to its promotion of collagen synthesis,local angiogenesis and reduction of inflammatory mediators. |
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