猪脱细胞真皮基质促进皮肤细胞迁移修复小鼠创面的研究  

Porcine acellular dermal matrix promotes the migration of skin cells to repair wounds in mice

作　　者：王知 李恭驰[2] 李炳辉[1] 杜烨 杨鸿[1] 周云华 潘银根[3] 冯自波[1] Wang Zhi;Li Gongchi;Li Binghui;Du Ye;Yang Hong;Zhou Yunhua;Pan Yingen;Feng zibo(Department of Wound Repair,Wuhan Liyuan Hospital,Tongji Medical College,Huazhong University of Science and Technology,Hubei Chronic Wound and Diabetic Foot Medical Clinical Research Center,Wuhan 430077,China;Department of Hand Surgery,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China;Department of Plastic and Cosmetic,Qidong People's Hospital,Qidong 226200,China)

机构地区：[1]华中科技大学同济医学院附属梨园医院创面修复科湖北省慢性创面及糖尿病足医学临床中心,武汉430077 [2]华中科技大学同济医学院附属协和医院手外科,武汉430022 [3]启东市人民医院整形美容科,启东226200

出　　处：《中华实验外科杂志》2020年第7期1230-1233,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年科学基金(81801922);国家重大疾病多学科合作诊疗能力建设项目(国卫办医函[2019]542号);湖北省自然科学基金面上项目(20I8CFB756);湖北省科技创新平台专项资助建设项目(2018BCC340)。

摘　　要：目的观察猪脱细胞真皮基质(pADM)对小鼠创面的修复作用及皮肤细胞迁移的影响。方法 2018年9月至2019年10月,对20只C57BL/6鼠(购自中国湖北省实验动物研究中心)背部用8 mm的皮肤活检器制作直径8 mm的全层皮肤缺损创面,直径8 mm、高3 mm一侧下端剪开45度,高1 mm窗口的柱状硅胶支撑架缝合于创面处形成创面窗。完全随机分成两组,一组创面窗中敷入pADM为pADM组,一组以纱布覆盖创面窗为对照组,给予及时护理。观察创面愈合情况,并对4周愈合创面组织进行苏木精-伊红(HE)染色。分离培养出生后2~3 d的乳鼠背部全皮层细胞,采用Transwell小室迁移实验检测pADM对皮肤细胞迁移的影响。4周创面组织进行毛囊干细胞标志物LGR5和角质形成细胞标志物角蛋白(K17)的免疫荧光染色。应用统计软件GraphPad Prism6分析,组间比较采用t检验。结果建模4周后,pADM组创面愈合率为(76.94±2.93)%,对照组创面愈合率为(57.59±4.08)%,与对照组比较,pADM可促进创面愈合(t=6.666,P<0.05)。4周创面组织HE染色结果显示pADM组在创面窗近窗口处有新生的毛囊生成,而对照组未见新生毛囊。Transwell结果显示对照组迁移细胞数为(52.67±1.45)个,pADM组迁移细胞数为(73.67±2.03)个,pADM可促进分离培养的皮肤细胞迁移(t=8.419,P<0.05)。4周创面组织免疫荧光染色结果显示pADM组创面窗组织中有毛囊干细胞的激活和角蛋白K17的表达。结论 pADM可促进创面愈合,并通过激活毛囊干细胞和角质形成细胞促进皮肤细胞迁移修复创面。Objective:To observe the repairing effect of porcine acellular dermal matrix(pADM)on wound and the effect on the migration of skin cells.Methods:From September 2018 to October 2019,twenty C57BL/6 mice(purchased from the Experimental Animal Research Center of Hubei Province,China)were used to make full-thickness skin defect wounds with 8 mm diameter by 8 mm skin biopsy device on the back.The lower end of the side with a diameter of 8 mm and a height of 3 mm was cut 45 degrees,and the cylindrical silica gel support frame with a height of 1 mm was sutured on the wound surface to form the wound window.Two groups were completely random divided:one group was treated with pADM in the wounds window,and one group was treated with none and gauze was used to cover the wounds.Timely nursing was given.The wound healing was observed and hematoxylin eosin(HE)staining was performed on the wound tissues after 4 weeks.The back cortex cells of suckling mice after 2-3 days of birth were isolated and cultured,and the effect of pADM on the migration of skin cells was detected by Transwell cell migration experiment.The 4-week wound tissues were stained with LGR5 and K17.The statistical software GraphPad Prism 6 was used for analysis,and t test was used for comparison between the two groups.Results:After 4 weeks of modeling,the wound healing rate in the pADM group was(76.94±2.93)%,and that in the control group was(57.59±4.08)%.Compared with the control group,pADM could promote wound healing(t=6.666,P<0.05).The HE staining showed that there were new hair follicles near the window of the wound in the pADM group,but there were no new hair follicles in the control group.Transwell results showed that the number of migration cells were 52.67±1.45 in control group and 73.67±2.03 in pADM group,pADM could promote the migration of cultured skin cells(t=8.419,P<0.05).The activation of hair follicle stem cells and the expression of keratin 17 were found in the wound window.Conclusion:pADM can promote wound healing,and promote the migration of s

关 键 词：脱细胞真皮基质 创面愈合 迁移 猪 

分 类 号：R641[医药卫生—外科学]