水溶性壳聚糖水凝胶对糖尿病小鼠感染全层皮肤缺损创面的作用及其机制  

Effects and mechanism of water-soluble chitosan hydrogel on infected full-thickness skin defect wounds in diabetic mice

作　　者：朱萌 陈禹州 区锦钊 李曌[2] 黄沙[2] 胡骁骅[3] 鞠晓燕 田野[1] 牛忠伟[1] Zhu Meng;Chen Yuzhou;Ou Jinzhao;Li Zhao;Huang Sha;Hu Xiaohua;Ju Xiaoyan;Tian Ye;Niu Zhongwei(Key Laboratory of Photochemical Conversion and Optoelectronic Materials,Technical Institute of Physics and Chemistry,Chinese Academy of Sciences,Beijing 100190,China;Research Center for Wound Repair and Tissue Regeneration,Medical Innovation Research Department,the PLA General Hospital,Beijing 100048,China;Department of Burns,Beijing Jishuitan Hospital,Beijing 100035,China)

机构地区：[1]中国科学院理化技术研究所中国科学院光化学转换与功能材料重点实验室,北京100190 [2]解放军总医院医学创新研究部创伤修复与组织再生研究中心,北京100048 [3]北京积水潭医院烧伤科,北京100035

出　　处：《中华烧伤与创面修复杂志》2022年第10期923-931,共9页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金面上项目(52073293)中国科学院理化技术研究所所长基金项目;中山市引进高端科研机构创新专项资金(2019AG003)。

摘　　要：目的探讨水溶性壳聚糖水凝胶对糖尿病小鼠感染全层皮肤缺损创面的作用及其机制。方法采用实验研究方法。采用循环冻融的方法制备由聚乙烯醇和明胶组成的对照水凝胶及由前述2种材料+水溶性壳聚糖组成的水溶性壳聚糖水凝胶。大体观察第1次冻融前后试管中2种敷料流动性,并比较12孔板中2种敷料最终形态的外观差异。取细胞株L929和HaCaT,均分别按照随机数字表法(分组方法下同)分为对照水凝胶组和水溶性壳聚糖水凝胶组,分别加入相应敷料培养24 h,采用细胞计数试剂盒8检测细胞增殖活力。取兔血红细胞悬液,分为生理盐水组、聚乙二醇辛基苯基醚(Triton X-100)组、对照水凝胶组和水溶性壳聚糖水凝胶组,分别作相应处理后孵育1 h,采用酶标仪检测红细胞的溶血程度。取24只11~14周龄雌性db/db小鼠,在其背部制作全层皮肤缺损创面并在创面处滴加耐甲氧西林金黄色葡萄球菌(MRSA)液,72 h后将小鼠分为空白对照组、磺胺嘧啶银水胶组、对照水凝胶组、水溶性壳聚糖水凝胶组,分别作相应处理。伤后0(即刻)、7、14、21 d,大体观察创面愈合情况并计算伤后14、21 d创面愈合率;伤后14 d,检测创面中MRSA浓度;伤后21 d,采用苏木精-伊红染色法对创面进行组织学分析,采用免疫荧光法检测创面中细胞CD31表达并计算其阳性百分率。取Raw264.7细胞,分为进行相应处理的白细胞介素4(IL-4)组、空白对照组、对照水凝胶组、水溶性壳聚糖水凝胶组,培养48 h,采用流式细胞仪检测细胞中CD206阳性细胞百分率。样本数均为3。对数据行独立样本t检验、单因素方差分析、重复测量方差分析、LSD检验及Dunnett T3检验。结果试管中2种敷料在进行冻融前都具有一定的流动性,冻融1次后均形成半固态凝胶。12孔板中2种敷料最终形态均基本呈稳定的半透明片状,透明度差异不大。培养24 h,水溶性壳�Objective To explore the effects and mechanism of water-soluble chitosan hydrogel on infected full-thickness skin defect wounds in diabetic mice.Methods The experimental research method was adopted.The control hydrogel composed of polyvinyl alcohol and gelatin,and the water-soluble chitosan hydrogel composed of the aforementioned two materials and water-soluble chitosan were prepared by the cyclic freeze-thaw method.The fluidity of the two dressings in test tube before and after the first freeze-thawing was generally observed,and the difference in appearance of the final state of two dressings in 12-well plates were compared.According to random number table(the same grouping method below),the cell strains of L929 and HaCaT were both divided into water-soluble chitosan hydrogel group and control hydrogel group,respectively.After adding corresponding dressings and culturing for 24 h,the cell proliferation activity was measured using cell counting kit 8.Rabbit blood erythrocyte suspensions were divided into normal saline group,polyethylene glycol octyl phenyl ether(Triton X-100)group,water-soluble chitosan hydrogel group,and control hydrogel group,which were treated accordingly and incubated for 1 hour,and then the hemolysis degree of erythrocyte was detected by a microplate reader.Twenty-four female db/db mice aged 11-14 weeks were selected,and full-thickness skin defect wounds on their backs were inflicted and inoculated with the methicillin-resistant Staphylococcus aureus(MRSA),72 h later,the mice were divided into blank control group,sulfadiazine silver hydrogel group,control hydrogel group,and water-soluble chitosan hydrogel group,which were treated accordingly.On post injury day(PID)0(immediately),7,14,and 21,the healing of the wound was observed.On PID 14 and 21,the wound healing rate was calculated.On PID 14,MRSA concentration in wounds was determined.On PID 21,the wounds were histologically analyzed by hematoxylin and eosin staining;the expression of CD31 in the wounds was detected by immunofluorescence met

关 键 词：水凝胶 壳聚糖 感染 皮肤 慢性创面 创面修复 

分 类 号：R587.2[医药卫生—内分泌]