| Background:Wound healing of skin defects is complex.For wounds with large wound area and deep damage depth,it is an excellent choice to accept artificial dermis grafting at the first stage surgery,and 2～3 weeks later,to accept autologous razor-thin skin grafting at the second stage surgery.In addition,the effectiveness of numerous cytokines such as fibroblast growth factor(FGF)on wounds healing has been widely researched,however,the traditional view is that direct external application or in vivo injection of exogenous FGFs may not achieve the desired therapeutic effect as the effective concentration cannot be maintained for a long time.Therefore,some teams have tried to integrate various them into skin substitutes for combined application.However,we believe that considering the current situation,it is still difficult to achieve mass production of these types of artificial dermis.Therefore,we manufactured a collagen-chondroitin sulfate(CS)scaffold material by imitating the artificial dermis materials on the market.Then,combined with recombinant human acidic fibroblast growth factor(rh-aFGF)in a single full dose during the first-stage artificial dermis transplantation,which is simple and completely feasible in the current clinical work,to explore whether this contributed to the healing of full-thickness skin defect wounds on the back of Balb/c-nu mice.Objective:1.To prepare 4 kinds of collagen-chondroitin sulfate scaffolds with mass fraction(wt.%)of 1%,0.8%,0.6%and 0.4%and to measure their physical properties.2.To investigate the cell compatibility and the cell adhesion properties of the 4 collagen-CS scaffolds in vitro.3.To select a collagen-CS scaffold with good physical properties,cell compatibility and cell adhesion properties to repair full thickness skin defect in Balb/c-nu mice combined with a single use of sufficient dose of recombinant human acidic fibroblast growth factor(rh-aFGF),and to evaluate its efficacy.Methods and results:1.4 kinds of collagen-CS scaffolds with 1%,0.8%,0.6%and 0.4%wt.%were successfully prepared by lyophilization and a number of physical and chemical properties were related to the mass fractions.2.Human adipose-derived stem cells(hADSCs)and human umbilical vein endothelial cells(HUVECs)were seeded on the 4 kinds of collagen-CS scaffolds with different mass fractions respectively.And it turned out all 4 kinds of collagen-CS scaffolds had good cell adhesion properties3.It was able to promote the proliferation of hADSCs and HUVECs seeded in 0.6%collagen-CS scaffolds to combine use of rh-aFGF within 24 hours.4.Performed the first stage surgery on Balb/c-nu mice that was full-thickness skin defect modeling and artificial dermis transplantation,combined use of rh-aFGF,and 9 days later,performed the second stage surgery of SD rat skin flap transplantation.And it turned out the skin transplantations’ survival rate on day 16 was improved and the wound contraction rates on day 16,21 and 28 were decreased.Conclusion:1.A number of physical and chemical properties are related to the mass fractions of collagen-CS scaffolds.Among them,with the decrease of the collagen-CS scaffold mass fraction,the average pore size of the collagen-CS scaffolds’ surface and bottom surface will increase.2.All 4 kinds of collagen-CS scaffolds have good cell compatibility and cell adhesion properties.For hADSCs and HUVECs,the collagen-CS scaffolds with larger pore size have shown better cell compatibility in vitro.3.It is able to promote the proliferation of hADSCs and HUVECs seeded in 0.6%collagen-CS scaffolds to combine use of rh-aFGF within 24 hours.4.A single and sufficient combined use of rh-aFGF during the first stage surgery can significantly improve the skin transplantations’ survival rate on day 16 and decrease the wound contraction rates on day 16,21 and 28 after the second stage surgery and to inhibit scarring in the full-thickness skin defect wounds of Balb/c-nu mice. |
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