红光联合银离子敷料治疗慢性难愈合创面的最佳方案筛选及机制

Screening and mechanism of the best treatment of red light and silver ion dressing for treatment of chronic non-healing wounds

背景:临床上多采用红光照射和银离子敷料治疗慢性难愈合创面,但是红光照射联合银离子敷料治疗慢性难愈合创面的最佳照射时间和不同银离子敷料的组合尚未确定。目的:探讨红光联合银离子敷料治疗大鼠慢性难愈合创面的最佳照射时间与敷料组合。方法:取SD大鼠,采用全层皮肤缺损+创面涂抹金黄色葡萄球菌+皮下注射氢化可的松的方式制作慢性难愈合创面模型。采用随机数字表法将造模成功的72只大鼠随机分为4组,每组18只:在创面标准换药的基础上分别给予红光照射20 min+脂质水胶体硫酸银敷料覆盖(A1B1组)、红光照射20 min+藻酸钙纤维银敷料覆盖(A1B2组)、红光照射30 min+脂质水胶体硫酸银敷料覆盖(A2B1组)、红光照射30 min+藻酸钙银纤维敷料覆盖(A2B2组),每24 h换药、照射一次并更换敷料。连续治疗14 d后,检测各组大鼠创面愈合率、细菌菌落数、炎症反应、组织形态学及血管新生情况。结果与结论:①随着治疗时间的延长,4组大鼠创面愈合率增加,A2B2组大鼠治疗后3,7,14 d的创面愈合率高于其他3组(P<0.05);②治疗后第7天的创面细菌培养结果显示,A2B2组细菌菌落数低于其他3组(P<0.05);Western-blotting检测显示,随着治疗时间的延长,4组大鼠创面组织中肿瘤坏死因子α、白细胞介素6的蛋白表达均下降,白细胞介素10的蛋白表达均升高,A2B2组肿瘤坏死因子α、白细胞介素6的蛋白表达低于其他3组(P<0.05),白细胞介素10的蛋白表达高于其他3组(P<0.05);③治疗后第14天的创面苏木精-伊红染色显示,A2B2组可见大量的胶原纤维平行分布且连接最紧密,明显优于其他3组;④治疗后第7天的创面免疫荧光染色显示,A2B2组CD31荧光强度表达高于其他3组(P<0.05);治疗后3,7,14 d的q-PCR检测显示,A2B2组血管内皮生长因子a、血管内皮生长因子受体2的mRNA表达均高于其他3组(P<0.05);治疗后3,7,14 d的Western-blotting检测显示,A2B2组血管内皮生长因子a、血管内皮生长因子受体2的蛋白表达均高于其他3组(P<0.05);⑤结果表明,红光照射30 min联合藻酸钙银纤维敷料治疗慢性难愈合创面的效果更优。

BACKGROUND:Red light irradiation and silver ion dressing are mostly used to treat chronic difficult healing wounds clinically,but the optimal irradiation time of red light irradiation and silver ion dressing for chronic non-healing wounds,and the combination of different silver ion dressings have not been determined.OBJECTIVE:To investigate the optimal irradiation time and dressing combination of red light and silver ion dressing in the therapy of chronic non-healing wounds.METHODS:The chronic non-healing wound model was made by applying Staphylococcus aureus on the whole skin defect and subcutaneous hydrocortisone injection in SD rats.72 rat models were randomly divided into 4 groups with 18 rats in each group by random number table method.The rats were treated on the basis of standard dressing change and the following therapy:A1B1 group(red irradiation 20 minutes+lipid hydrocolloidal silver sulfate dressing),A1B2 group(red light irradiation 20 minutes+calcium alginate fiber dressing),A2B1 group(red light irradiation 30 minutes+lipid hydrocolloidal silver sulfate dressing),and A2B2 group(red light irradiation 30 minutes+calcium alginate fiber dressing);change dressing,irradiate once,and change dressing every 24 hours.After 14 days of continuous treatment,wound healing rate,bacterial colony number,inflammatory response,histomorphology and angiogenesis were detected in each group.RESULTS AND CONCLUSION:(1)With the extension of treatment time,the wound healing rate of rats in the four groups was increased,and the wound healing rate of rats in the A2B2 group at 3,7,and 14 days after treatment was higher than that in the other three groups(P<0.05).(2)The wound bacterial culture results on day 7 after treatment demonstrated that the number of bacterial colonies in the A2B2 group was lower than that in the other three groups(P<0.05).Western blot assay exhibited that with the extension of treatment time,the protein expressions of tumor necrosis factorαand interleukin-6 in wound tissue of rats in the four groups were decreased,while the protein expressions of interleukin-10 were increased.The protein expressions of tumor necrosis factorαand interleukin-6 in the A2B2 group were lower than those in the other three groups(P<0.05).The protein expression of interleukin-10 in the A2B2 group was higher than that of the other three groups(P<0.05).(3)The wound hematoxylin-eosin staining on day 14 after treatment demonstrated that a large number of collagen fibers in the A2B2 group were parallel distributed and the most closely connected,which was significantly better than the other three groups.(4)The results of immunofluorescence staining indicated that the fluorescence intensity expression of CD31 in the A2B2 group was higher than that in the A1B1,A1B2 and A2B1 groups(P<0.05).q-PCR detection at 3,7,and 14 days after treatment exhibited that the mRNA expressions of vascular endothelial growth factor a and vascular endothelial growth factor receptor 2 in the A2B2 group were higher than those in the other three groups(P<0.05).Western blot assay at 3,7 and 14 days after treatment revealed that the protein expressions of vascular endothelial growth factor a and vascular endothelial growth factor receptor 2 in the A2B2 group were higher than those in the other three groups(P<0.05).(5)These findings confirm that 30 minutes of red light irradiation combined with silver alginate fiber dressing has better results in treatment of chronic non-healing wounds.