| ObjectiveTo study the effects of chitosan/antiphlogistic traditional Chinese medicine and the combination of the two on the healing process of mouse skin wounds.Analyze the corresponding relationship between different drug groups/different concentration groups and mouse skin wound healing effects.Further discuss the mechanism of chitosan on promoting wound healing and provide experimental basis for the clinical application of chitosan trauma dressings.Method1.Animal grouping and modelingUsing 306 healthy Kunming inbred male mice,300 of them were randomly divided into 9 treatment groups and a blank control group,30 mice in each group.The remaining 6 were normal group.A round skin full-cut wound of approximately 0.5 cm2 was made on each mouse's back,All traumatized mice were treated immediately after the preparation of the skin defect wounds for 2 consecutive days for 9 days.The 9 treatment groups were high,medium and low(8mg/L,4mg/L,2mg/L)chitosan trauma dressing treatment groups,high,medium and low(8mg/L,4mg/L,2mg/L).Chitosan quaternary ammonium trauma dressing treatment group,4mg/L chitosan +1/2/antiphlogistic traditional Chinese medicine trauma dressing treatment group,4mg/L chitosan quaternary ammonium + 1/2/antiphlogistic traditional Chinese medicine treatment group and/antiphlogistic traditional Chinese medicine trauma dressing treatment group.2.Mouse Wound Shrinkage EvaluationOn the 1st,3rd,5th,7th and 9th days after the operation,the wounded parts of the mice were standardized photography.The wounds of each mouse wound were recorded and computer image processing was performed using ImageJ image processing software.According to the data distribution characteristics,ANOVA analysis of variance was performed on the data of wound area shrinkage of each group on the 1st,3rd,5th,7th and 9th days after surgery.3.The expression of IL-1,TNF-a,TGF-? and PDGF determined by ELISAOn the 1st,3rd,5th,7th and 9th days after operation,6 mice were randomly selected from each group to take blood from the eyeball.The blood is centrifuged on the same day to collect the supernatant and the expression of IL-1,TNF-?,TGF-?,and PDGF were measured by enzyme-linked immunosorbent assay kits.4.Histopathological evaluationOn the 3rd,5th,7th,and 9th days after surgery,the blood-collected mice were decapitated and sacrificed.Approximately 1cm2 of the wound tissue sample wastaken surgically.Tissue samples were made in paraffin sections and stained with HE.The histopathological changes were observed under an ordinary light microscope.RESULTSOn the first day after operation,there was no significant difference in the contraction rate of wounds between the groups(P>0.05).On the 3rd and 5th days after operation,wond shrinkage of the high and middle chitosan group?the high and middle chitosan quaternary ammonium group?the chitosan medium +1/2traditional Chinese medicine group?the chitosan quaternary ammonium + 1/2traditional Chinese medicine group?the antiphlogistic traditional Chinese medicine group was significantly faster than the blank group(P<0.05).The high-concentration group had a faster wound shrinkage than the low-concentration group(P<0.05).There was no significant difference in the shrinkage rate of wounds between chitosan group and chitosan quaternary ammonium group(P>0.05).There was no significant difference in the contraction rate of wound between the combination group and high concentration group(P>0.05).On the 7th and 9th days after operation,the wounds of all treatment groups contracted significantly faster than the blank group(P<0.05).Histopathological analysis revealed that:Compared with the control group,the treatment group had less inflammation,faster epithelial cell proliferation,rich neovascularization,and mature granulation tissue.Compared with the low-dose group,the high-concentration group had an earlier period of inflammation reduction,faster proliferation of epithelial cells,an earlier appearance of granulation tissue,and a shorter time for complete skin restoration.The degree of recovery under the microscope was similar between the chitosan group and the chitosan quaternary ammonium group.The degree of recovery under the combined application group was similar to that of the high concentration group,and the complete skin structure was seen on the 9th day after operation.The expression of IL-1 and TNF-a in the treatment group was significantly lower than that in the blank group(P<0.01).The expression of IL-1 and TNF-a in the high concentration group was lower in the lower concentration group(P<0.05).There was no significant difference(P>0.05)in the expression of IL-1 and TNF-a in chitosan quaternary ammonium group?The expression of IL-1 in the combination group was lower(P<0.05),and the expression of TNF-a was not significantly different(P>0.05)from the high concentration group.The expression of TGF-? and PDGF in the treatment group was significantly higher than that in the blank group(P<0.05).The expression of TGF-? and PDGF in the high concentration group was higher in the lower concentration group(P<0.05).There was no significant difference in the expression of TGF-? and PDGF between chitosan group and chitosan quaternary ammonium group(P>0.05).The expression of PDGF was higher in the combination group than in the higher concentration group(P<0.05).There was no statistical difference in expression(P>0.05).Conclusion1.Chitosan,chitosan quaternary ammonium,and anti-inflammatory and/antiphlogistic traditional Chinese medicine have the effects of reducing inflammation and promoting the expression of growth factors.Thereby they can all promote wound healing.2.Compared with the low-concentration,the high-concentration chitosan and chitosan quaternary ammonium have a better effect of reducing inflammation and promoting growth factor expression.As a result high-concentration chitosan and chitosan quaternary have a stronger ability to promote wound healing.3.The abilitys of chitosan and chitosan quaternary ammonium to reduce inflammation and promote growth factors are approximately the same,and their ability to promote wound healing are similar.4.The combination of chitosan/chitosan quaternary ammonium and antiphlogistic traditional Chinese medicine have the best ability to reduce inflammatory response and promote the expression of growth factors.Therefor they have the best ability of promote wound healing. |
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