Study On The Adhesion And Mechanism Of Porcine Fibrin Sealant In Wound

Objective: Stitching is often used for the closure of wounds in clinical operations,but it is not only difficult to apply to fragile organs,hard to prevent leakage of tissue fluid but easy to cause hypertrophic scar hyperplasia.Especially with the progress of healing,quickly and effectively promote the closure of the wound while reducing the formation of scar has become an important concern.Since 1998,the FDA has approved the use of fibrin sealant(FS)in the United States.And then it is widely used clinically for closure and adhesion,while new products are constantly being introduced for new indications.With the advancement of medicine and the increasing rigor of dressings,fibrin seleant with degradability and good biocompatibility have become one of the first choices for wound closure.However,the main clinical indications of the domestically marketed fibrin seleant are all blood effects rather than adhesion.Therefore,basing on the successful development of a new type of porcine fibrin seleant,the seleant can be applied to the skin.A series of studies on the effect and mechanism of wound healing,in order to determine the effect of the adhesive on skin healing,preliminary verification of its adhesion,and provide experimental basis for the clinical application of the adhesive in skin wound treatment.Methods: 1.The CCK-8 method was used to detect the cytotoxicity of porcine fibrin sealant on L929 and HUVECs.The cytotoxicity of porcine fibrin glue was further verified by microscopic observation of cell growth morphology.Hoechst/PI double staining was used to further investigate the effect of porcine fibrin sealant on apoptosis of L929 and HUVECs.2.Different treatments were given by constructing a 4 cm long full-slice knife model on the skin at the back of SD rats.Through the healing strength of the healing site,HE staining and Masson staining,the healing effect of the porcine fibrin sealant was investigated.3.Explore the mechanism of porcine fibrin sealant promoting wound healing by molecular and protein levels by Q-PCR and Western Blot.Result: 1.The porcine fibrin sealant extract were applied to L929 and HUVECs cells for 24 h,48 h and 72 h,respectively.The cell viability of L929 cells were 84.64±1.66,81.97±0.31 and 83.98±2.42,respectively.It was lower than the control group(P<0.05).The cell viability of porcine fibrin sealant extract on HUVECs at 24 h,48 h and 72 h was 98.76±6.07,96.80±1.88 and 88.74±2.21,respectively.The HUVECs cells treated with the extract had no significant effect on the cells for 24 h and 48 h,but the cell viability was lower than that of the control group at 72 h(P<0.05).The cytotoxicity of the extract was determined according to the standard,and the porcine fibrin sealant had a cytotoxicity of 0-1 grade and was less toxic.It was further confirmed by microscopic observation that the porcine fibrin sealant had no effect on the cell growth morphology of L929 and HUVECs.The effect of porcine fibrin sealant on apoptosis was further verified.After treated with porcine fibrin sealant extract for 24 h,the apoptosis of L929 and HUVECs was not obvious,and there was no significant difference with the control group.It is further suggested that the porcine fibrin sealant is less toxic and has good biocompatibility.2.By establishing the model of back skin wound healing in SD rats,the wound healing rate was preliminarily judged by the tensile strength of the healing tissue.The results showed that porcine fibrin sealant promoted the wound healing at the beginning of the healing stage.Next,HE and Masson staining of the wound healing site confirmed that the porcine fibrin sealant can promote epidermal re-epithelialization and collagen deposition to promote wound healing.3.Using Q-PCR to investigate the effect of porcine fibrin sealant on VEGFA mRNA,bFGF mRNA,Collagen I mRNA and TGF-?1 mRNA expression in promoting skin healing.Western Blot was used to investigate the effects of porcine fibrin sealant on the expression of VEGFA,bFGF,Collagen I,TGF-?1 and ?-SMA protein in healing wounds.The experimental results showed that the Q-PCR results were almost consistent with the results of Western Blot,porcine fibrin sealant promoted wound healing by up-regulating bFGF,VEGFA and ?-SMA in the early stage of healing,and down-regulated the expression of Collagen I and TGF-?1,which are related to scar.During the healing remodeling period,the fibrin sealant group promptly promoted the apoptosis of myofibroblasts and decreased the expression of ?-SMA protein.Conclusion: Porcine fibrin sealant has good biocompatibility and can promote skin wound healing while reducing scar formation.