| Skin trauma is a very common disease in veterinary clinical practice.After skin trauma,the blood at the wound site oozes out,which can lead to inflammatory response and local infection,and systemic infection such as pus and sepsis in severe cases.Studies have shown that bacterial cellulose has the effect of promoting wound healing,and carvol has antibacterial,anti-inflammatory,anti-tumor,antioxidant and other biological activities,but the use of bacterial cellulose and parsley phenol as a composite membrane form in veterinary clinic for trauma treatment has not been reported.Therefore,the purpose of this experiment is to prepare bacterial cellulose/parsley phenol composite membrane and observe its clinical effect on the skin wound healing of dogs,so as to provide an alternative wound dressing for the skin wound healing of dogs in veterinary clinic.In this experiment,the solution soaking method was adopted.BC was impregnated in carvacrol ethanol solution with different concentrations,and the adsorption of BC to carvacrol was saturated by ultrasonic shock.After ethanol removal,the BC/carvacrol composites with different concentrations were obtained.The minimum bactericidal concentration(MBC)of carvol in the composite membrane material was determined by bacteriostatic ring and plate counting method,and the composite membrane material containing different concentrations of carvol was prepared according to the MBC.The physical characteristics of bacterial cellulose and composite membrane materials containing different concentrations of carvol were tested.The rat skin wound model was established and randomly divided into control group and three different concentration composite membrane experimental groups to screen the best concentration of composite membrane in the process of skin wound healing.The skin wound model was established with dogs as experimental animals and randomly divided into control group,BC group and composite membrane group.After modeling,BC group and composite membrane group changed dressing once a day,observed and recorded the wound healing every day,and determined the time of complete wound healing.Take photos to record the wound healing at the 0,7,14,21 and 28 days,and measure the wound area through Image J software to analyze the wound healing rate.The pathological sections of skin were made for histopathological observation.Western blot(WB)was used to detect Wnt signal pathway and growth factor;Elisa kit was used to detect inflammatory factors;Real-time fluorescence quantitative PCR was used to detect the expression of cytokine m RNA related to wound healing.The results showed that the MBC of carvecol in the composite film material was 31mg/m L,and according to MBC,the composite film material was prepared with the concentration of carvecol at 31mg/m L,62mg/m L and 93mg/m L.The characterization results meet the requirements of wound dressing.The average skin wound healing rate after moulding was significantly higher in the experimental group than in the control group(P<0.05).At the 3rd day,the mean wound healing rate was significantly higher in the 62 mg/m L and 93 mg/m L experimental groups than in the control group(P<0.05).On the 7th,10 th and 14 th day,the average wound healing rate of 62mg/m L and 93mg/m L experimental group was significantly higher than that of control group(P<0.01)There was no significant difference between 93 mg/m L and 62 mg/m L groups(P>0.05).In terms of mean healing time,the 93 mg/m L and 62 mg/m L experimental groups were superior to the control group(P<0.01).Based on the consideration of drug toxicity and cost,62 mg/m L was selected as the drug concentration of the composite membrane for the formal experiment.The animal skin irritation test showed that the composite membrane prepared in this study did not irritate the skin.WB experiment showed that compared with the control group,carvacrol significantly increased the expression of Wnt5 a protein,promoted the phosphorylation of Gsk3-β,and caused the significant expression of VEGF growth factor.Meanwhile,carvacrol significantly down-regulated the expression of NF-κB signaling pathway.Elisa kit showed that carvacrol significantly inhibited the expression of inflammatory factors compared with control group.Compared with the control group,the expression of growth factors in the composite membrane group was significantly increased,indicating that carvacrol can improve the expression of m RNA in the skin wound tissue.In conclusion,BC/carvavol composite membrane can promote skin wound repair and accelerate wound healing by regulating Wnt/β-catenin and NF-κB signaling pathways. |
