摘要
背景:生物工程支架是临床治疗慢性难愈合性创面的常用方法,具有一定的疗效,但也存在一些问题。脂肪来源干细胞作为新兴的治疗慢性难愈合性创面的方法具有其独特的疗效和优势。但是,脂肪来源干细胞和生物工程支架联合应用治疗慢性难愈合性创面的疗效和机制尚不清楚。目的:研究脂肪来源干细胞联合胶原蛋白生物工程支架移植对慢性难愈性创面血管内皮生长因子表达的影响。方法:实验共选取31只SD大鼠,购自上海斯莱克实验动物责任有限公司。取1只大鼠腹股沟处脂肪,分离培养原代脂肪来源干细胞。将另外30只SD大鼠随机分为对照组、模型组、支架组、脂肪来源干细胞组和联合组。对照组制备全层皮肤缺损创面(在大鼠腰椎正中偏上部两侧分别制备1处全层皮肤缺损伤口),术后每日常规换药;其余各组制备2处慢性难愈合性创面(同对照组制备全层皮肤缺损伤口,创面局部注射醋酸氢化可的松);模型组术后每日给予常规换药;支架组术后给予生物蛋白工程支架覆盖创面;脂肪来源干细胞组给予自体自体脂肪干细胞移植;联合组在脂肪来源干细胞移植后给予生物工程支架覆盖创面。干预7d后取材,观察创面情况,测量创面面积大小;采用免疫组化法检测血管内皮生长因子表达,采用Westernblot检测血管内皮生长因子蛋白表达量,采用qP CR检测血管内皮生长因子mR NA表达情况。结果与结论:(1)对照组创面面积显著小于其他组创面面积(P <0.05);联合组创面面积显著小于模型组、支架组和脂肪来源干细胞组(P<0.05);(2)对照组血管内皮生长因子表达水平最高,血管内皮生长因子蛋白和mR NA表达水平均显著高于其他组(P <0.05);联合组血管内皮生长因子表达较高,血管内皮生长因子蛋白和mR NA表达水平显著多于模型组、支架组和脂肪来源干细胞组(P <0.05);(3)结果提示,脂肪来源干细胞联合胶原蛋白生物工程支架移植上调难愈合性创面血管内皮生长因子表达,促进创面愈合。
BACKGROUND:Bioengineering scaffolds are commonly used in the treatment of chronic refractory wounds and have achieved some effects,but there are also some problems.Adipose-derived stem cells(ADSCs)as a new method for the treatment of chronic refractory wounds have its unique efficacy and advantages.However,the efficacy and mechanism of ADSCs combined with bioengineering scaffolds in the treatment of chronic refractory wounds are not yet clear.OBJECTIVE:To study the effect of ADSCs combined with autologous collagen biomaterial scaffold transplantation on the expression of vascular endothelial growth factor(VEGF)during the healing of chronic refractory wounds.METHODS:Thirty-one Sprague-Dawley rats were purchased from Shanghai Slack Laboratory Animals Co.,Ltd.Adipose tissues from the groin of a rat were harvested,and the primary ADSCs were isolated and cultured in vitro.The remaining 30 rats were randomly divided into normal control group,model group,collagen biomaterial scaffold group(Plenac group),autologous ADSCs transplantation group(ADSCs group)and autologous ADSCs+collagen biomaterial scaffold group(ADSCs+Plenac group),6 rats in each group.In the normal control group,full-thickness skin wounds in the central and upper part of the bilateral lumbar vertebrae were made,followed by daily routine dressing.In the other groups two chronic refractory wounds were made in the central and upper part of the bilateral lumbar vertebrae followed by local injection of hydrocortisone acetate.Model group were given daily routine treatment;Plenac group treated with fibrin scaffold to cover the wound,followed by daily routine dressing;ADSCs group treated with autologous ADSCs transplantation followed by daily routine dressing;ADSCs+Plenac group received autologous ADSCs transplantation and bioengineering scaffold implantation to cover the wound,followed by daily routine dressing.After 7 days of intervention,all the rats were executed.The following procedures were subsequently carried out:observing wound situation,measuring wound size,detecting the expression of VEGF by immunohistochemical method,detecting the expression of VEGF protein using western blot assay,and detecting the expression of VEGF mRNA by qPCR.RESULTS AND CONCLUSION:(1)The wound area in the control group was smaller than that in the other groups(P<0.05),while the wound area in the ADSCs+Plenac group was also smaller than that in the model,Plenac and ADSCs groups(P<0.05).(2)The expression of VEGF at protein and mRNA levels was higher in the control group compared with the other groups(P<0.05),while the protein and mRNA expression of VEGF in the ADSCs+Plenac group was significantly higher than that in the model,Plenac and ADSCs group(P<0.05).To conclude,ADSCs combined with the collagen bioengineering scaffold can upregulate VEGF expression in wound healing,and promote healing of wound healing.
作者
庄兢
杨宇
丁力
郑清健
Zhuang Jing;Yang Yu;Ding Li;Zheng Qing-jian(Fujian Provincial Hospital,Fuzhou 350001,Fujian Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第33期5274-5280,共7页
Chinese Journal of Tissue Engineering Research
基金
福建省自然科学基金(2017J0105)~~
