脂肪间充质干细胞复合人工真皮修复大鼠深度烧伤创面的研究  

Adipose-derived mesenchymal stem cells combined with PELNAC for repairing deep burn wound in rats

作　　者：范友芬[1] 乐欣[1] 晋国营[1] 虞耀华 

机构地区：[1]宁波市第二医院烧伤科,315000

出　　处：《浙江医学》2017年第19期1633-1637,1732,共6页Zhejiang Medical Journal

基　　金：浙江省医药卫生科技计划(2013KYB234)

摘　　要：目的探讨脂肪间充质干细胞(ADSCs)复合人工真皮(皮耐克)修复大鼠深度烧伤创面的作用。方法用胶原酶法消化SPF级绿色荧光蛋白转基因大鼠腹股沟脂肪,获得ADSCs,用流式细胞仪分析鉴定其表面特异性标志物,并观察ADSCs向成骨细胞、成脂细胞分化的潜能。建立大鼠深度烧伤模型,将大鼠对称的左、右创面分为实验组和对照组。实验组采用人工真皮+ADSCs治疗,对照组采用人工真皮+0.9%氯化钠注射液治疗。分别于术后7、14、21d观察创面最大直径以评估创面愈合情况,术后21d时荧光显微镜下观察创面冷冻切片评估实验组与对照组创面情况。结果成功获得大鼠ADSCs,具有较强的增殖能力及多向分化能力。ADSCs对间充质干细胞标志物CD29、CD90、CD44呈现高表达;对造血干细胞表面标志物CD34、白细胞共同抗原CD45则呈低表达。治疗后14d时,实验组创面最大直径明显小于对照组(P<0.05),即实验组创面愈合速度比对照组明显增快。治疗后21d,实验组创面可见散在点状绿色荧光,而对照组未观察到明显荧光。结论 ADSCs联合人工真皮作用于深度烧伤创面可显著促进创面愈合,是一种有效的治疗深度烧伤的方法。Objective To assess the efficacy of adipose-derived mesenchymal stem cells（ADSCs） combined with artificial dermis（PELNACR） for repairing deep burn wound in rats.Methods ADSCs were isolated from adult SPF grade green fluorescent protein-transgenic rats.The specific cell surface markers of ADSC were analyzed with flow cytometry,the differentiation of ADSCs to osteoblasts was induced and the multilineage differentiation potential was observed.The third-degree burn wound model was induced on the back of Spregue-Dawley（SD） rats.The model rats were randomly divided into two groups：rats in test group（n=20） received ADSCs ＋PELNACRfor wound repairing and those in control group（n=20） received saline ＋ PELNACR.The maximum diameter of wound on d7,d14 and d21 after burns was measured,respectively.On the d21 the skin samples of burn area were taken to evaluate wound healing by frozen section.Results The ADSCs were successfully isolated,cultured,and identified.The ADSCs had the ability to proliferate continuously and showed characteristics of multi-potent mesenchymal stem cells.The results of flow cytometry showed that CD44,CD90 and CD29 were highly expressed in ADSCs,while the hematopoietic cell surface marker CD34 and leukocyte common antigen CD45 were lowly expressed.On the d14 after burns,the maximum diameter of the wound was significantly lower in test group than that in control group（P〈0.05）.On d21 after burns,fluorescence microscope showed scattered fluorescence was distributed in ADSCs-transplanted wound tissue in test group,but not in control group.Conclusion The combination of ADSCs with artificial dermis（PELNACR） can significantly accelerate wound healing,indicating that it might be an effective method for treating deep burn.

关 键 词：深度烧伤 人工真皮 脂肪间充质干细胞 

分 类 号：R644[医药卫生—外科学]