| Background:Diabetes foot ulcer is one of the most common complications in diabetes patients,and it is difficult to treat.Research has found that platelet-rich plasma(PRP)contains various high concentrations of growth factors,which play an important role in promoting wound healing and other aspects.However,there are a large number of proteases and inflammatory factors in diabetes wounds,which can degrade growth factors and cytokines of PRP,leading to a significant reduction in the effect of PRP.In recent years,some scholars have reported that neutrophil extracellular traps(NETs)play an important role in diabetes wounds.The high concentration of NETs,its components MPO,NE,histone,etc.,can degrade the extracellular matrix in the wound tissue,aggravate the wound inflammatory reaction,and delay the wound healing of diabetes.If we first reduce the formation of NETs in diabetes wounds,and then inject PRP,will the effect be better?There are currently no relevant reports.Objective: Observe the impact of wound healing after deoxyribonuclease 1(Dnase1)is injected into diabetes wound to degrade neutrophil extracellular traps(NETs),platelet-rich plasma(PRP)is injected together and explore its mechanisms.Methods: Four ulcer wound models were established on diabetic rats and randomly divided into four groups: Group A(Dnase + PRP group),Group B(PRP group),Group C(Dnase group)and Group D(control group).Group A(Dnase + PRP group): The wounds were rinsed with normal saline.According to the standard of 15 mg/kg,the total amount of Dnase1 injected into each rat was evenly divided into 8 equal parts.Eight equal points on the wound edge of the rat were selected,and each equal part of Dnase1 was evenly injected subcutaneously with a 1ml syringe.At the same time,the center of two Dnase1 injection points on the edge of the wound was selected as the PRP injection point,a total of 8 points,and each point was evenly injected with 0.1ml PRP,and the wounds were coated with vaseline gauzes and medical semi-permeable films.Group B(PRP group):The wounds were rinsed with normal saline.PRP was uniformly injected subcutaneously at 8 points on the edge of the wound by the same method,and the wounds were coated with vaseline gauzes and medical semi-permeable films.Group C(Dnase group): Washed the wounds with normal saline,Dnase1 was injected in the same way,then the wounds were covered with vaseline gauzes and medical semi-permeable films.Group D(control group): The wounds were rinsed with normal saline,normal saline was uniformly injected subcutaneously into the wound by PRP injection method,then the wounds were covered with vaseline gauzes and medical semi-permeable films.Take 12 rats on the 5 and 14 days after injection,observed the wound condition,the wound healing rate and the growth of granulation tissue,and the sections were stained by HE staining immunohistochemistry method for the neovascularization,the infiltration of fibroblast and inflammatory cell.Tissue growth factor,inflammatory factor and NETs related indexe were detected and compared.Results: 1)In two observations,more fresh granulation tissue、neovascularization and fibroblast were observed,but the number of neutrophil infiltration was decreased in Dnase+PRP group.2)The wound healing rates of Dnase+PRP group,PRP group,Dnase group and control group were 92.59±2.13%,76.23±3.72%,70.34±5.79 %and 55.68±3.29%,and the highest healing rate was Dnas + PRP group(P <0.05).3)The concentration of growth factor VEGF and TGF-β1 in the wound tissue of Dnase+PRP group and PRP group were higher than those in the other two groups.and the difference was significant(P<0.05).4)In the wound tissue of Dnase+PRP and Dnase groups,the production of myeloperoxidase(MPO)and elastase(NE),the markers of neutrophil extracellular traps formation was decreased(P<0.05),the release of pro-inflammatory factors such as tumor necrosis factor-α(TNF-α)was decreased(P<0.05),and the release of anti-inflammatory factors such as Interleukin-8(IL-8)and Interleukin-10(IL-10)was increased(P<0.05).Conclusions:1.Dnase1 can reduce the level of NETs in wound granulation tissue of diabetic rats;2.PRP can promote the growth of granulation tissue and epithelialization of wound tissue in diabetic rats after the reduction of NETs level in wound tissue;3.The combined injection of Dnase1 and PRP can produce a synergistic effect: reduce the level of pro-inflammatory factors in the wound of diabetic rats and improve the chronic persistent inflammation of the wound.At the same time,increase the concentration of growth factors,promote cell proliferation,angiogenesis,and promote wound healing. |
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