低频电磁场在基因修饰表皮干细胞移植修复创面中的作用  

Effects of low-frequency electromagnetic fields on skin wound healing after the transplantation of gene modified epidermal stem cells

作　　者：梁玲[1] 李新平[2,3] 白文芳[2,3] 白利明[2,3] 朱洪翔[2,3] 许伟成[2,3] 冯玉[4] 王鑫[1] 陈艺[4] 张鸣生[1,2,3] 

机构地区：[1]暨南大学第一临床学院,广东省广州市510630 [2]广东省人民医院康复医学科 [3]广东省老年医学研究所,广东省广州市510080 [4]南方医科大学,广东省广州市510515

出　　处：《中国组织工程研究》2013年第32期5827-5833,共7页Chinese Journal of Tissue Engineering Research

基　　金：广东省自然科学基金(S2012010009498);广东省自然科学基金(S2012010009308)~~

摘　　要：背景:烧伤、慢性创面等大面积皮肤缺损的创面修复是临床上亟待解决的难题。表皮干细胞和角质化细胞生长因子可为创面的治疗提供新的策略。低频电磁场作为一种非侵入性物理刺激在创面修复中已被认为较佳的方法。目的:探讨低频电磁场在角质化细胞生长因子基因修饰的表皮干细胞移植促进小鼠皮肤全层缺损创面修复中的作用。方法:体外分离培养SD乳鼠表皮干细胞,用5-溴脱氧尿核苷进行标记,成功转染角质化细胞生长因子基因腺病毒表达载体后,移植于昆明小鼠全层缺损创面。建立全层缺损创面小鼠模型,分别进行表皮干细胞移植,角质化细胞生长因子修饰的表皮干细胞移植,角质化细胞生长因子修饰的表皮干细胞移植外加低频电磁场干预。结果与结论:移植后第9,16天,低频电磁场干预的角质化细胞生长因子基因修饰的表皮干细胞移植组创面收缩率优于其他各组(P<0.05)。移植后第9天,免疫荧光染色法检测显示,各组组创面中均有5-溴脱氧尿核苷阳性细胞分布。移植后第16天,Westernblot检测显示,低频电磁场干预的角质化细胞生长因子基因修饰的表皮干细胞移植组和角质化细胞生长因子基因修饰的表皮干细胞移植组创面组织中角质化细胞生长因子蛋白表达高于表皮干细胞移植组(P<0.05);移植后第16,30天,苏木精-伊红染色结果显示,低频电磁场干预的角质化细胞生长因子基因修饰的表皮干细胞移植组的创面组织上皮化现象较明显。结果证实,低频电磁场有促进角质化细胞生长因子基因修饰的表皮干细胞移植修复小鼠皮肤全层缺损创面的作用。BACKGROUND： The repair and management of full-thickness skin defects resulting from burns and chronic wounds remain a significant unmet clinical challenge. Using epidermal stem cells and keratinocyte growth factor for full-thickness wound repair is a promising approach. Low-frequency electromagnetic fields which are a non-invasive physical stimulation therapy have been recognized as a good method to enhance wound healing. OBJECTIVE： To develop a new strategy to accelerate wound healing by transplanting transfected epidermal stem cells and keratinocyte growth factor and treating with low-frequency electromagnetic fields in a mouse model. METHODS： Epidermal stem cells from Sprague-Dawley neonatal rats were isolated and cultured in vitro, then the cells were labeled with 5-bromo-2-deoxyuridine and transfected by Ad-KGF, a recombinant adenovirus carrying the keratinocyte growth factor. Mice were given to create full thickness skin wound on the dorsum and randomly assigned to four groups： control group, transplantation of epidermal stem cells group, transplantation of keratinocyte growth factor gene modified epidermal stem cells group, and transplantation of keratinocyte growth factor gene modified epidermal stem cells plus low-frequency electromagnetic field exposure group. RESULTS AND CONCLUSION： The best healing pattern was observed in the keratinocyte growth factor gene modified epidermal stem cells plus low-frequency electromagnetic field exposure group （P 0.05） at days 9 and 16. 5-Bromo-2-deoxyuridine labeled cells existed in the wound in the treated groups at day 9. A significantly increased expression of endogenous keratinocyte growth factor was detected in the transplantation of Keratinocyte Growth Factor gene modified epidermal stem cells group, and transplantation of keratinocyte growth factor gene modified epidermal stem cells plus low-frequency electromagnetic field exposure group at day 16. A well-advanced epithelialization was observed in transplantation of keratinocyte growth factor gene

关 键 词：干细胞 干细胞移植 低频电磁场 表皮干细胞 角质细胞生长因子 皮肤全层缺损创面 创面愈合 省级基金 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]