负载锌离子的复合水凝胶对糖尿病小鼠全层皮肤缺损感染创面的作用及机制  

Effects and mechanisms of zinc ion-loaded composite hydrogel on infected full-thickness skin defect wounds in diabetic mice

作　　者：潘泽平 石云龙 袁志强 彭毅志 安中莲 乐率[3] 龚雅利 Pan Zeping;Shi Yunlong;Yuan Zhiqiang;Peng Yizhi;An Zhonglian;Le Shuai;Gong Yali(Institute of Burn Research,State Key Laboratory of Trauma and Chemical Poisoning,the First Affiliated Hospital of Army Medical University(the Third Military Medical University),Chongqing 400038,China;Emergency Department,Jiangjin Hospital Affiliated to Chongqing University,Chongqing 402260,China;Department of Microbiology,College of Basic Medical Sciences,Key Laboratory of Microbial Engineering Under the Educational Committee in Chongqing,Army Medical University(the Third Military Medical University),Chongqing 400038,China)

机构地区：[1]陆军军医大学(第三军医大学)第一附属医院全军烧伤研究所,创伤与化学中毒全国重点实验室,重庆400038 [2]重庆大学附属江津医院急诊科,重庆402260 [3]陆军军医大学(第三军医大学)基础医学院微生物学教研室,微生物工程重庆市高校重点实验室,重庆400038

出　　处：《中华烧伤与创面修复杂志》2024年第9期866-875,共10页Chinese Journal of Burns And Wounds

基　　金：重庆英才项目(CQYC20220511656)。

摘　　要：目的探讨负载锌离子的复合水凝胶(以下简称含锌水凝胶)对糖尿病小鼠全层皮肤缺损感染创面的作用及其机制。方法该研究为实验研究。制备聚(甘油癸二酸酯)-共-聚乙二醇-共聚-邻苯二酚/季铵化壳聚糖水凝胶(以下简称单纯水凝胶)和在单纯水凝胶的基础上添加锌离子的疏松多孔且具有良好黏附性的固态含锌水凝胶。计算用磷酸盐缓冲液(PBS)浸泡14 d后含锌水凝胶中锌离子的释放率。检测用单纯水凝胶、含锌水凝胶、PBS培养2 h后耐甲氧西林金黄色葡萄球菌(MRSA)浓度。利用酶标仪检测单纯水凝胶、含锌水凝胶、PBS对1,1-二苯基-2-三硝基苯肼(DPPH)的清除率,反映清除氧自由基能力。测量用单纯水凝胶、含锌水凝胶、PBS培养24 h后人脐静脉内皮细胞(HUVEC)形成的血管长度。采用细胞计数试剂盒8检测用单纯水凝胶、含锌水凝胶、PBS培养24 h后L929细胞的活力。将小鼠红细胞悬液分为用PBS处理的空白对照组和用相应溶液处理的单纯水凝胶组、含锌水凝胶组和Triton X-100组,利用酶标仪检测孵育2 h后红细胞的溶血情况并计算溶血率。以上实验样本数均为3。取21只6~8周龄雄性C57BL/6J小鼠,在背部脊柱对称位置各制备1个用MRSA感染的全层皮肤缺损创面。将小鼠分为滴加PBS的空白对照组和用相应水凝胶处理的单纯水凝胶组和含锌水凝胶组。伤后3 d,检测空白对照组、单纯水凝胶组、含锌水凝胶组小鼠创面中的细菌浓度,样本数为4。于伤后0(即刻)、3、7、14 d,大体观察小鼠创面感染情况并计算伤后3、7、14 d创面愈合率,样本数为5。伤后14 d,行苏木精-伊红染色和Masson染色分别检测小鼠创面中的新生上皮和胶原生成情况,行免疫荧光染色检测小鼠创面中血管新生及M2型巨噬细胞分布情况。结果浸泡14 d后,含锌水凝胶中锌离子的释放率为(70.5±4.6)%。与用含锌水凝胶比较,用PBS和单�Objective To investigate the effects and mechanisms of zinc ion-loaded composite hydrogel(hereinafter referred to as the zinc-containing hydrogel)on infected full-thickness skin defect wounds in diabetic mice.Methods This study was an experimental study.A poly(glycerol sebacate)-co-poly(ethylene glycol)-g-catechol prepolymer/quaternized-chitosan hydrogel(hereinafter referred to as the simple hydrogel)and a solid-state zinc-containing hydrogel with porous and good adhesion by adding zinc ions to the simple hydrogel were prepared.The release rate of zinc ions from the zinc-containing hydrogel after immersion in phosphate buffer solution(PBS)for 14 days was calculated.The concentration of methicillin-resistant Staphylococcus aureus(MRSA)cultured for 2 hours with the simple hydrogel,zinc-containing hydrogel,and PBS was measured.The scavenging ability of the simple hydrogel,zinc-containing hydrogel,and PBS for 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl(DPPH)was detected using microplate reader to reflect the ability of oxygen free radical removal.The length of vessels formed by human umbilical vein endothelial cells(HUVECs)cultured for 24 hours with the simple hydrogel,zinc-containing hydrogel,and PBS was measured.The cell viability of L929 cells cultured for 24 hours with the simple hydrogel,zinc-containing hydrogel,and PBS was detected using the cell counting kit-8.The mouse red blood cell suspension was divided into blank control group treated with PBS,simple hydrogel group,zinc-containing hydrogel group,and Triton X-100 group treated with corresponding solution.Hemolysis was detected using microplate reader after 2 hours of treatment,and the hemolysis rate was calculated.All experiments had a sample size of 3.Twenty-one C57BL/6J mice aged 6-8 weeks were taken,and a full-thickness skin defect wound was prepared in the symmetrical position on the back spine and infected with MRSA.Mice were divided into blank control group treated with PBS,simple hydrogel group,and zinc-contai

关 键 词：生物相容性材料 水凝胶 感染 糖尿病 实验性 锌离子 抗菌性 创面修复 

分 类 号：R641[医药卫生—外科学] R587.2[医药卫生—临床医学] R318.08