直流电场刺激脂肪来源干细胞修复糖尿病大鼠难愈性创面  

Effect of adipose-derived stem cells stimulated by direct current electric field on refractory wound healing in diabetic rats

作　　者：张锐 刘兰 谢德富 林映汐 任红静 颜洪 Zhang Rui;Liu Lan;Xie Defu;Lin Yingxi;Ren Hongjing;Yan Hong(Department of Plastic and Burn Surgery Affiliated Hospital of Southwest Medical University National Key Clinical Construction Specialty Wound Repair and Regeneration Lab oratory,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属医院整形烧伤外科,国家临床重点建设专科,创面修复与再生实验室,四川省泸州市646000

出　　处：《中国组织工程研究》2023年第10期1484-1491,共8页Chinese Journal of Tissue Engineering Research

基　　金：(泸州市-西南医科大学)合作项目(2019):PHD2-HIF信号途径改善糖尿病性创面愈合的作用研究-重点项目(2019LZXNYDZ08),项目负责人:颜洪;西南医科大学校级科研项目:近红外探针吲哚箐绿(ICG)体外标记脂肪间充质干细胞及其在体成像的实验研究(2021ZKQN074),项目负责人:刘兰。

摘　　要：背景:脂肪来源干细胞在促进创面修复和再生方面潜力巨大,课题组前期已证明直流电场可以促进脂肪来源干细胞的增殖并且不改变其干细胞的生物学特性。目的:探究直流电场刺激大鼠脂肪来源干细胞对糖尿病大鼠难愈性创面修复的影响。方法:(1)获取大鼠脂肪来源干细胞,鉴定后分别给予0,100mV/mm直流电场刺激预处理,频率为1 h/d,连续3 d;(2)构建糖尿病难愈性创面大鼠模型并随机分为3组,每组7只,共21只,分别在创面局部多点注射1 mL直流电场刺激和未刺激的脂肪来源干细胞,细胞浓度为1×10^(9) L,PBS组给予等量的PBS;(3)造模后3,7,14 d,大体观察创面愈合程度;激光散斑血流对比成像仪记录大鼠创面血流灌注情况;苏木精-伊红染色评价创面愈合组织学变化并进行微血管计数;免疫组化观察创面组织血管内皮生长因子的表达;免疫荧光观察创面组织CD31的表达。结果与结论:(1)与未刺激脂肪来源干细胞组和PBS组相比较,直流电场刺激脂肪来源干细胞组在第14天时创面接近完全愈合,创面愈合速度明显加快(P <0.05);(2)直流电场刺激脂肪来源干细胞组在第3天时即开始出现创面上皮化,第14天时表皮结构较其余2组更加完整和连续,创面血管生成增加(P <0.05),血管内皮生长因子和CD31表达增加明显(P <0.05);(3)3组大鼠创面血流灌注随着时间进展都出现逐渐达到峰值再回调的现象,直流电场刺激脂肪来源干细胞组治疗后不同时间点血流灌注较PBS组显著增加(P <0.05),并在第7天时,明显高于未刺激脂肪来源干细胞组(P <0.05);(4)结果说明直流电场作用后的大鼠脂肪来源干细胞可以加速糖尿病难愈性创面愈合进程,其潜在的机制可能与其增加血流灌注、促进血管内皮生长因子表达和血管生成数量增加有关,这为体外大量扩增脂肪来源干细胞,加快干细胞的临床转化提供了新的思路。BACKGROUND:Adipose-derived stem cells have a great potential in wound healing and regeneration and we have proven that a direct-current electric-field can promote the proliferation of human adipose-derived stem cells without changing biological characteristics.OBJECTIVE:To investigate the effect of rat adipose-derived stem cells stimulated by a direct-current electric-field on refractory wound healing in diabetic rats.METHODS:(1) Rat adipose-derived stem cells were obtained.After identification,they were pretreated with 100 mV/mm or 0 mV/mm of direct-current electric-field stimulation,with a frequency of 1 h/d,for 3 days.(2) Diabetic refractory wound models were constructed and randomly divided into three groups,with 7 rats in each group,totaling 21 rats.Animals in the electric-field-rat adipose-derived stem cell and rat adipose-derived stem cell groups were injected with 1 mL of electric-field-rat adipose-derived stem cells or normal rat adipose-derived stem cells,respectively,at multiple points of the wound,with a cell concentration of 1×10^(9) cells/L.The PBS group was given the same amount of PBS.(3) At 3,7,and 14 days after model establishment,the degree of wound healing was observed.Laser speckle contrast imaging was used to record the blood perfusion of rat wounds.The histological changes of wound healing were evaluated by hematoxylin and eosin staining.Microvessels were counted.The expression of vascular endothelial growth factor in the wound tissue was observed by immunohistochemistry.The expression of CD31 was observed by immunofluorescence.RESULTS AND CONCLUSION:(1) Compared the normal rat adipose-derived stem cells and PBS groups,wounds in the electric-field-rat adipose-derived stem cell group were nearly completely healed on day 14,and the wound-healing speed was significantly accelerated(P < 0.05).(2) In the electric-field-rat adiposederived stem cell group,wound epithelization began on day 3.On day 14,the epidermal structure was more complete and continuous in the electric-field-rat adipose-derive

关 键 词：直流电场 脂肪来源干细胞 细胞增殖 创面愈合 糖尿病 大鼠 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R622