| Objective: 1.The skin of mice was burned with an electric cauterizer,and the wounds were infected with Staphylococcus aureus to establish mouse wound healing and hypertrophic scar models 2.To explore the therapeutic effect of Crocin Ⅰ and Cuttlebone alone or in combination on wound healing and scar formation,so as to provide experimental basis for clinical treatment of wound healing and prevention of scar formation.Methods: 1.A more stable wound healing and scar model in mice was established by electric burn combined with bacterial infection.Wound healing rate,HE staining,Masson staining,immunohistochemical staining and Elisa assay were used to evaluate the histopathological changes of wound skin.2.Crocin Ⅰ,Cuttlebone,Crocin Ⅰ + Cuttlebone were used to treat the wounds of mice infected by electric burn combined with bacteria.The therapeutic effects were evaluated by calculating the wound healing rate,HE,Masson,and immunohistochemical staining.Results: Part Ⅰ: 1.Wound healing rate:The wound healing time of the model group was longer,and the wound healing rate of each phase was lower than that of the control group.2.The histopathological results of the wound: HE staining showed that the wound healing of Model group progressed slowly.On the 7th day,there was no skin tissue in the model group,but a small amount of connective tissue appeared between the scab and the muscular layer in the control group.Compared with the Control group,the intact epidermis and dermis were not found on the 14 th day and at 21 days,the complete epidermal layer was formed.3.Masson staining and collagen volume fraction(CVF) analysis: The Control group and the Model group showed varying degrees of collagen accumulation after wound healing.With the continuous progress of wound repair,the CVF of the Model group was significantly higher than that of the Control group at the later stage.4.Immunohistochemical staining and average optical density analysis: The expression of IL-6,TGF-β1,α-SMA and Col Ⅲ in the model group was higher than that in the control group during the whole process of wound healing;Compared with the control group,the expression of TGF-β3 in the Model group was lower on the 7th day and increased with the process of wound healing.The expression of COLⅠA1 was significantly higher at the later stage of wound healing.5.ELISA was used to detect the concentration of IL-6 in venous blood: the content of IL-6 in venous blood of the Model group was higher than that of the Control group.Part II: 1.Wound healing rate: The wound healing rate of the treatment group was higher than that of the Model group at each time point,and the wound healing rate was faster.2.The histopathological results of the wound:HE staining showed that the wound healing of each treatment group progressed faster,and the skin structural integrity was stronger than that of the Model group on the 7th day.On day 14,the epidermis and dermis became thinner.3.Masson staining and collagen volume fraction(CVF)analysis:On day 7,CVF of the CROⅠ group and CROⅠ+CB group was higher than that of the Model group.The CVF of each group increased on the 14 th day,but the CVF of each treatment group was lower than that of the Model group until the 28 th day.Immunohistochemical staining and average optical density analysis:Compared with the model group,the synthesis of IL-6 in CROⅠ group was lower,and the synthesis of IL-6 CROⅠ+CB group was lower over time;in principle,the expression of TGF-β1 in wound tissue of each treatment group was lower than that of the model group during the whole process of wound repair;compared with the model group,the secretion of TGF-β3 decreased significantly on the 7th day,and TGF-β3 synthesis increased significantly on the 21 st day in CROⅠ group.COLⅢ in the treatment group was lower than that in the model group in the whole process of wound healing,while α-SMA and COLⅠA1 were lower in the middle and later stage.Conclusion: 1.After artificial inoculation of Staphylococcus aureus,the skin inflammatory response of the wound is enhanced,the integrity of skin structure is poor,and the scar structure similar to human pathological scar is formed.2.Both Crocin I and Cuttlebone can promote wound healing and reduce the production of pathological scar in mice infected by Staphylococcus aureus. |
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