聚六亚甲基双胍/甘油凝胶敷料通过清除细菌生物膜和增加Toll样受体2/4的表达以促进慢性创面愈合

Polyhexamethylene biguanide/glycerol gel promotes chronic wound healing by inhibiting bacterial biofilm formation and increasing the expression of TLR2/4

目的探讨聚六亚甲基双胍/甘油胶(PHMB/Glycerol gel)在慢性创面治疗中的应用效果,及其对创面内细菌生物膜和肉芽组织生长相关蛋白表达的影响。方法于48只BALB/c小鼠背部制作一2 cm×2 cm皮肤创面,随机分为PHMB/Glycerol gel组、银离子敷料(Silver ion)组、传统纱布敷料(control)组,每组16只。治疗10 d和12 d后,取创面处组织进行苏木精-伊红(HE)染色观察肉芽组织形态以及计算毛细血管数量。并进行蛋白质印迹法(Western blot)和免疫组织化学分析(IHC)检测促进炎症的Toll样受体2(TLR2)和Toll样受体4(TLR4)等的表达。结果PHMB/Glycerol gel组伤口愈合时间要显著短于Silver ion组和control组[(21.000±1.549)d比(25.000±1.773)、(31.000±1.195)d,F=32.71,P<0.05]。Western blot的结果显示PHMB/Glycerol gel组TLR2(126283±8675比65470±3106、27290±3698,F=227.60,P<0.001)和TLR4(135698±10130比58488±3376、19152±3730,F=247.30,P<0.001)的表达水平高于其他组。IHC实验结果显示PHMB/Glycerol gel组的TLR(47.330±1.783比24.940±8.226、17.010±5.028,F=23.15,P<0.01)和TLR4的表达密度(45.160±3.957比30.940±4.842、16.060±4.989,F=29.77,P<0.05)均显著高于Silver ion和control组。结论PHMB/Glycerol凝胶敷料可通过提高TLR2、TLR4的表达来促进细菌生物膜的清除和肉芽组织的生长,是慢性感染性创面修复的理想选择。

Objective To investigate the effect of PHMB/glycerol gel on chronic wounds and related mechanisms,such as bacterial biofilms and some molecular factors that influence the growth of granulation tissue.Methods A 2 cm×2 cm skin wound was prepared on the back of 48 BALB/c mice and randomly divided into PHMB/glycerol gel group,Silver ion group,and traditional gauze dressing(control)group,with 16 mice in each group.After 10 and 12 days,48 of the skin defects were subjected to hematoxylin and eosin(HE)staining,and the morphology of the granulation tissue(capillary blood vessels)was observed.Western blotting and immunohistochemical(IHC)analyses were performed to explore the expression of Toll-like receptors 2(TLR2),Toll-like receptors 4(TLR4).Results The wound healing time in PHMB/glycerol gel group was significantly shorter than that in Silver ion group and control group[(21.000±1.549)d vs.(25.000±1.773),(31.000±1.195)d,F=32.71,P<0.05,respectively].Western blotting results showed that the expression levels of TLR2(126283±8675 vs.65470±3106,27290±3698,F=227.60,P<0.001,respectively)and TLR4(135698±10130 vs.58488±3376,19152±3730,F=247.30,P<0.001,respectively)in PHMB/glycerol gel group were higher than in Silver ion group and control group.IHC results showed that the expression levels of TLR2(47.330±1.783 vs.24.940±8.226,17.010±5.028,F=23.15,P<0.01,respectively)and TLR4(45.160±3.957 vs.30.940±4.842,16.060±4.989,F=29.77,P<0.05,respectively)in PHMB/glycerol gel group were significantly higher than those in Silver ion group and control group.Conclusion PHMB/glycerol gel can promote bacterial biofilm clearance and granulation tissue growth by increasing TLR2 and TLR4 expression,making it an ideal choice for chronic infectious wound repair.