摘要
目的:探讨骨髓间充质干细胞(Bone mesenchymal stem cells,BMSCs)的外泌体对表皮细胞(HaCaT细胞)增殖、侵袭与迁移的影响,以及对小鼠烧伤创面愈合的作用。方法:分离培养小鼠的BMSCs,超速离心法收集BMSCs来源的外泌体,采用BMSCs来源的外泌体培养HaCaT细胞,CCK-8法检测BMSCs来源的外泌体对HaCaT细胞增殖的影响,细胞划痕实验和Transwell实验检测BMSCs来源的外泌体对HaCaT细胞迁移与侵袭的影响。选取25只C57BL/6小鼠构建烧伤模型,22只成功建模,将存活小鼠分为对照组和外泌体组,每组11只,外泌体组烫伤周围注射BMSCs来源的外泌体,对照组小鼠同时注射等体积PBS溶液。采用Image J软件分析小鼠创面愈合率,通过HE染色观察创面组织学变化,采用酶联免疫吸附法(ELISA)检测创面组织中炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)的水平,免疫组织化学染色检测创面组织Ⅰ型胶原(ColⅠ)和Ⅲ型胶原(ColⅢ)表达。结果:成功分离培养出小鼠BMSCs,并得到BMSCs来源的外泌体。与对照组比较,实验组HaCaT细胞增殖活性明显升高;细胞的划痕愈合率较对照组显著升高,细胞侵袭数目较对照组也显著增加,差异均具有统计学意义(P<0.05)。与对照组小鼠比较,经BMSCs来源的外泌体处理的小鼠,第3、7、14、21、28天的创面愈合率显著升高,创面组织结构得到明显改善,且组织中促炎因子TNF-α、IL-1β的含量均显著降低,而抗炎因子IL-10的含量显著升高,Ⅲ型胶原表达明显,差异均具有统计学意义(P<0.05)。结论:BMSCs外泌体可促进HaCaT细胞的增殖、侵袭与迁移能力,并减轻小鼠烧伤创面组织的炎症程度,促进创面愈合。
Objective To investigate the effect of exosomes secreted by bone marrow mesenchymal stem cells(BMSCs)on the proliferation,invasion and migration of human keratinocytes(HaCaT cells),and on the healing of burn wounds in mice.Methods BMSCs from mice were isolated and cultured,exosomes derived from BMSCs were collected by ultracentrifugation,HaCaT cells were cultured with exosomes derived from BMSCs,and the effect of exosomes derived from BMSCs on the proliferation of HaCaT cells was detected by CCK-8 method.Scratch test and Transwell test were used to detect the influence of BMSCs-derived exosomes on HaCaT cell migration and invasion.Twenty-five C57BL/6 mice were selected to construct a burn model.After successful modeling(n=22),the surviving mice were divided into a control group and an exosome group,with 11 in each group.Exosomes derived from BMSCs were injected around the scald in the exosomes group,and mice in the control group were injected with an equal volume of PBS solution at the same time.Image J software was used to analyze the wound healing rate of mice,and the histological changes of the wound were observed by HE staining.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-10(IL-10)in wound tissue.Immunohistochemical staining was used to detect the expression of typeⅠcollagen(ColⅠ)and typeⅢcollagen(ColⅢ)in wound tissue.Results The mouse BMSCs were successfully isolated and cultured,and exosomes derived from BMSCs were obtained.Compared with the control group,the HaCaT cell proliferation activity in the experimental group was significantly increased;the scratch healing rate of the cells was significantly higher than that of the control group,and the number of cell invasion was also significantly higher than that of the control group.The differences were statistically significant(P<0.05).Compared with control mice,mice treated with BMSCs-derived exosomes had a significantly higher woun
作者
唐强
黄志群
廖秋姣
陈端凯
陆钢
姜艳
唐乾利
TANG Qiang;HUANG Zhi-qun;LIAO Qiu-jiao;CHEN Duan-kai;LU Gang;JIANG Yan;TANG Qian-li(Department of Burn Plastic Surgery and Wound Repair Surgery,Affiliated Hospital of Youjiang Medical College for Nationalities,Baise 533000,Guangxi,China;Department of Spine and Bone Surgery,Affiliated Hospital of Youjiang Medical College for Nationalities,Baise 533000,Guangxi,China;Youjiang Medical College for Nationalities,Baise 533000,Guangxi,China)
出处
《中国美容医学》
CAS
2021年第9期1-6,共6页
Chinese Journal of Aesthetic Medicine
基金
国家自然科学基金面上项目(编号:81774327)
“‘广西特聘专家’专项经费资助”
“广西医学高层次领军人才培养‘139’计划项目资助”。
