CAJ | 학술논문

背景:基因药物与干细胞联合应用促进难治性大面积创面修复.目的:观察携带肝细胞生长因子基因的重组腺病毒(adenovirus hepatocyte growth factor,Ad-HGF)修饰的间充质干细胞对大鼠皮肤放射性损伤创面促愈合的作用.方法:体外分离、培养雄性Wistar大鼠骨髓间充质干细胞,转染Ad-HGF后备用.40只雌性Wistar大鼠用X射线局部照射制备急性放射性皮肤损伤模型.并将模型大鼠随机分为单纯间充质干细胞治疗组、单纯 Ad-HGF 治疗组、Ad-HGF修饰的间充质干细胞治疗组、溶剂对照组4组,制备模型即刻各组将相应细胞悬液、病毒悬液及溶剂行创面边缘多点注射.结果与结论:苏木精-伊红染色结果显示,伤后21 d Ad-HGF修饰的间充质干细胞治疗组再生表皮明显薄于溶剂对照组,并可见丰富的新生小血管,且表皮较单纯间充质干细胞治疗组和单纯Ad-HGF治疗组更平整,新生小血管更丰富;免疫组织化学结果表明,Ad-HGF修饰的间充质干细胞治疗组创面中肝细胞生长因子表达均强于其他组;羟脯氨酸含量Ad-HGF修饰的间充质干细胞治疗组在7 d后显著低于其他组(P <0.05);原位杂交结果显示,Ad-HGF修饰的间充质干细胞治疗组修复的皮肤组织中性别决定基因sry表达强于单纯间充质干细胞治疗组.结果提示,Ad-HGF修饰的间充质干细胞对皮肤放射性损伤创面有显著的促愈合修复作用,且优于二者单独应用.
배경:기인약물여간세포연합응용촉진난치성대면적창면수복.목적:관찰휴대간세포생장인자기인적중조선병독(adenovirus hepatocyte growth factor,Ad-HGF)수식적간충질간세포대대서피부방사성손상창면촉유합적작용.방법:체외분리、배양웅성Wistar대서골수간충질간세포,전염Ad-HGF후비용.40지자성Wistar대서용X사선국부조사제비급성방사성피부손상모형.병장모형대서수궤분위단순간충질간세포치료조、단순 Ad-HGF 치료조、Ad-HGF수식적간충질간세포치료조、용제대조조4조,제비모형즉각각조장상응세포현액、병독현액급용제행창면변연다점주사.결과여결론:소목정-이홍염색결과현시,상후21 d Ad-HGF수식적간충질간세포치료조재생표피명현박우용제대조조,병가견봉부적신생소혈관,차표피교단순간충질간세포치료조화단순Ad-HGF치료조경평정,신생소혈관경봉부;면역조직화학결과표명,Ad-HGF수식적간충질간세포치료조창면중간세포생장인자표체균강우기타조;간포안산함량Ad-HGF수식적간충질간세포치료조재7 d후현저저우기타조(P <0.05);원위잡교결과현시,Ad-HGF수식적간충질간세포치료조수복적피부조직중성별결정기인sry표체강우단순간충질간세포치료조.결과제시,Ad-HGF수식적간충질간세포대피부방사성손상창면유현저적촉유합수복작용,차우우이자단독응용.
BACKGROUND:Combined application of gene therapy and stem cel s can promote the healing of refractory large scale wound. OBJECTIVE:To observe the effect of adenovirus hepatocyte growth factor (Ad-HGF) modified bone marrow mesenchymal stem cel s (BMSCs) on radioactive skin wound healing. METHODS:The BMSCs were in vitro isolated and cultured from male Wistar rats and transfected with Ad-HGF. Forty female Wistar rats were irradiated using X-ray therapy machine to establish acute skin radiation damage model. The models were randomly divided into four groups:BMSCs treatment group, Ad-HGF treatment group, Ad-HGF modified BMSCs treatment group and control group. After irradiating, the suspension of BMSCs, Ad-HGF and Ad-HGF modified BMSCs suspension were injected directly into the radioactive wounds. RESULTS AND CONCLUSION:Hematoxylin-eosin staining results showed that at 21 hours after injury, the epidermis in Ad-HGF modified BMSCs treatment group was significantly thinner than that in the control group, and the epidermis in the Ad-HGF modified BMSCs treatment group was more regular than that in the BMSCs treatment group and Ad-HGF treatment group, many smal vessels were observed in the derma of radioactive wounds. Immunohistochemistry results showed that the expression of hepatocyte growth factor in Ad-HGF modified BMSCs treatment group was stronger than that in the other three groups;the content of hydroxyproline in the Ad-HGF modified BMSCs treatment group was significantly lower than that in the other three groups after 7 days (P<0.05);in situ hybridization showed that the expression of sex-determining sry gene in Ad-HGF modified BMSCs treatment group was stronger than that in the BMSCs treatment group. It demonstrates that Ad-HGF modified BMSCs can promote the healing of radioactive skin wound, and the effect is better than simple application of the suspension.