CAJ | 학술논문

　　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，Western blot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。
　　배경：소상、만성창면등대면적피부결손적창면수복시림상상극대해결적난제。표피간세포화각질화세포생장인자가위창면적치료제공신적책략。저빈전자장작위일충비침입성물리자격재창면수복중이피인위교가적방법。목적：탐토저빈전자장재각질화세포생장인자기인수식적표피간세포이식촉진소서피부전층결손창면수복중적작용。방법：체외분리배양SD유서표피간세포，용5-추탈양뇨핵감진행표기，성공전염각질화세포생장인자기인선병독표체재체후，이식우곤명소서전층결손창면。건립전층결손창면소서모형，분별진행표피간세포이식，각질화세포생장인자수식적표피간세포이식，각질화세포생장인자수식적표피간세포이식외가저빈전자장간예。결과여결론：이식후제9，16천，저빈전자장간예적각질화세포생장인자기인수식적표피간세포이식조창면수축솔우우기타각조(P<0.05)。이식후제9천，면역형광염색법검측현시，각조조창면중균유5-추탈양뇨핵감양성세포분포。이식후제16천，Western blot검측현시，저빈전자장간예적각질화세포생장인자기인수식적표피간세포이식조화각질화세포생장인자기인수식적표피간세포이식조창면조직중각질화세포생장인자단백표체고우표피간세포이식조(P<0.05)；이식후제16，30천，소목정-이홍염색결과현시，저빈전자장간예적각질화세포생장인자기인수식적표피간세포이식조적창면조직상피화현상교명현。결과증실，저빈전자장유촉진각질화세포생장인자기인수식적표피간세포이식수복소서피부전층결손창면적작용。
BACKGROUND:The repair and management of ful-thickness skin defects resulting from burns and chronic wounds remain a significant unmet clinical chal enge. Using epidermal stem cel s and keratinocyte growth factor for ful-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 cel s and keratinocyte growth factor and treating with low-frequency electromagnetic fields in a mouse model. METHODS:Epidermal stem cel s from Sprague-Dawley neonatal rats were isolated and cultured in vitro, then the cel s 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 ful thickness skin wound on the dorsum and randomly assigned to four groups:control group, transplantation of epidermal stem cel s group, transplantation of keratinocyte growth factor gene modified epidermal stem cel s group, and transplantation of keratinocyte growth factor gene modified epidermal stem cel s 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 cel s plus low-frequency electromagnetic field exposure group (P<0.05) at days 9 and 16. 5-Bromo-2-deoxyuridine labeled cel s 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 cel s group, and transplantation of keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group at day 16. A wel-advanced epithelialization was observed in transplantation of keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group at days 16 and 30. These results suggest that low-frequency electromagnetic fields enhanced wound healing fol owing the transplantation of keratinocyte growth factor gene modified epidermal stem cel s.