不同途径应用人脐带间充质干细胞外泌体治疗小鼠全层皮肤缺损创面的效果  

Effects of applying human umbilical cord mesenchymal stem cell exosomes through different pathways to treat full-thickness skin defect wounds in mice

作　　者：王宏宇 巴特[1] 周彪[1] 闫增强[1] 王睿甲 刘玲英[3] Wang Hongyu;Ba Te;Zhou Biao;Yan Zengqiang;Wang Ruijia;Liu Lingying(Department of Burns,Baogang Hospital of Inner Mongolia,Baotou 014010,China;Graduate School of Inner Mongolia Medical University,Hohhot 010059,China;Department of Nutrition,the Fourth Medical Center,PLA General Hospital,Beijing 100037,China)

机构地区：[1]内蒙古包钢医院烧伤科,包头014010 [2]内蒙古医科大学研究生院,呼和浩特010059 [3]解放军总医院第四医学中心营养科,北京100037

出　　处：《中华烧伤与创面修复杂志》2024年第4期314-322,共9页Chinese Journal of Burns And Wounds

基　　金：重大疾病防治科技行动计划创伤修复专项项目(2018ZX-01S-001S01);内蒙古自治区自然科学基金面上项目(2020MS08022);内蒙古自治区科技创新引导项目(CXYD2020BT03)。

摘　　要：目的探讨通过创面局部涂抹、创缘皮下注射和尾静脉注射人脐带间充质干细胞(hUCMSC)外泌体治疗小鼠全层皮肤缺损创面的效果,探究应用hUCMSC外泌体治疗创面的最佳给药途径。方法该研究为实验研究。从3名25~35岁于内蒙古包钢医院妇产科正常分娩产妇弃用脐带组织中提取hUCMSC外泌体并成功鉴定。选用120只6~8周龄雄性BALB/c小鼠,于其背部制备全层皮肤缺损创面后,按随机数字表法分为对照组(不进行给药处理)、创面局部涂抹组、创缘皮下注射组、尾静脉注射组(每组30只小鼠),分别通过创面局部涂抹、创缘皮下注射、尾静脉注射给予后3组小鼠0.2 mL含200μg hUCMSC外泌体的磷酸盐缓冲液。伤后7、14、21 d,观察创面的大体情况并计算创面愈合率;采集创面组织,分别通过苏木精-伊红和Masson染色观测病理学变化和胶原纤维情况,行CD31免疫组织化学染色观测新生微血管数量,采用酶联免疫吸附测定法检测肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的含量。各组各时间点样本数均为10。结果伤后7、14、21 d,4组小鼠创面均逐步愈合,其中创缘皮下注射组小鼠创面愈合情况最佳;3个给药组小鼠创面愈合率均显著高于对照组(P<0.05),创缘皮下注射组及尾静脉注射组小鼠创面愈合率均显著高于创面局部涂抹组(P<0.05),创缘皮下注射组小鼠创面愈合率均显著高于尾静脉注射组(P<0.05)。伤后7、14、21 d,与对照组比较,3个给药组小鼠创面组织生长、上皮化的速度显著加快,创面胶原纤维生成数量更多且排列更整齐。伤后7、14、21 d,在每200倍视野下,创面局部涂抹组小鼠创面组织新生微血管数量分别为(24.1±2.5)、(50.7±4.1)、(44.2±2.3)根,创缘皮下注射组小鼠创面组织新生微血管数量分别为(32.2±2.9)、(67.5±4.9)、(53.6±3.7)根,尾静脉注射组小鼠创面组织新生微血管数量分别为(27.8±2.4)、(59.1±3.7)、(4Objective To investigate the effects of human umbilical cord mesenchymal stem cell(hUCMSC)exosomes in the treatment of full-thickness skin defect wounds in mice through local wound application,subcutaneous injection at the wound margin,and tail vein injection,and to explore the optimal administration route of hUCMSC exosomes for wound treatment.Methods This study was an experimental study.hUCMSC exosomes were extracted from the discarded umbilical cord tissue of three normal delivery women aged 25-35 years in the Department of Obstetrics and Gynecology of Baogang Hospital of Inner Mongolia and successfully identified.Totally 120 male BALB/c mice aged 6-8 weeks were selected,and full-thickness skin defect wounds were prepared on the back of them.According to the random number table,the injured mice were divided into control group(without drug administration),local wound application group,wound margin subcutaneous injection group,and tail vein injection group(with 30 mice in each group).Mice in the latter three groups were given 0.2 mL phosphate buffer solution containing 200μg hUCMSC exosomes by local wound application,subcutaneous injection at the wound margin,and tail vein injection,respectively.On post injury day(PID)7,14,and 21,the general condition of the wound was observed,and the wound healing rate was calculated;the wound tissue was collected,the pathological changes and collagen fibers were observed respectively by hematoxylin-eosin staining and Masson staining,the number of new microvessels was observed by CD31 immunohistochemical staining,and the content of tumor necrosis factorα(TNF-α)and interleukin-6(IL-6)was detected by enzyme-linked immunosorbent assay.The sample number was 10 in each group at each time point.Results On PID 7,14,and 21,the wounds of mice in the 4 groups all healed gradually,and the wound healing of the mice in wound margin subcutaneous injection group was the best;the wound healing rates of mice in the three administration groups were significantly higher than those in control g

关 键 词：创伤和损伤 皮肤 间质干细胞 外泌体 炎症 新生血管化 病理性 创面修复 给药途径 

分 类 号：R641[医药卫生—外科学]