脂肪间充质干细胞膜片移植促进创面愈合的可行性

Feasibility of adipose-derived mesenchymal stem cell sheet transplantation for wound healing

背景:脂肪间充质干细胞因其能旁分泌多种生长因子、获取方便、具有免疫调节等作用,常被用于组织修复和功能重建。食管内镜黏膜下剥离后狭窄的预防和治疗是困扰医生和患者的难题。脂肪间充质干细胞结合新型组织工程技术——细胞膜片,具有促进创面修复、预防食管狭窄发生的潜能和应用价值。目的:探讨脂肪间充质干细胞膜片的生物学特征,分析其预防食管内镜黏膜下剥离后狭窄的可行性。方法:(1)将使用温度敏感培养皿制备的绿色荧光蛋白标记的脂肪间充质干细胞膜片设为实验组,常规贴壁脂肪间充质干细胞为对照组,通过ELISA、反转录实时PCR及Western blot比较2组血管内皮生长因子、转化生长因子β1、肝细胞生长因子含量;(2)取裸鼠制备皮肤人工溃疡,随机分为贴膜组与模型组,贴膜组立即在皮肤创面处贴附绿色荧光蛋白标记的脂肪间充质干细胞膜片。结果与结论:(1)脂肪间充质干细胞膜片中细胞含量较高,细胞外基质丰富,细胞与细胞之间连接存在;膜片组血管内皮生长因子、转化生长因子β1、肝细胞生长因子表达水平均高于对照组;(2)裸鼠背部脂肪间充质干细胞膜片移植成功,且贴膜组愈合速度快于模型组;(3)实验结果提示脂肪间充质干细胞结合细胞膜片技术预防食管内镜黏膜下剥离后狭窄具有可行性,但仍需大量的基础及临床研究进一步证实。

BACKGROUND:Adipose-derived mesenchymal stem cells are often used for tissue repair and functional reconstruction due to their paracrine actions of multiple growth factors,easy access and immunomodulatory effects.The prevention and treatment of esophageal stenosis after esophageal endoscopic submucosal dissection is a challenge for doctors and patients.Adipose-derived mesenchymal stem cell transplantation combined with the cell sheet technology has the potential to promoting wound repair and preventing esophageal stenosis.OBJECTIVE:To investigate the biological characteristics of adipose-derived mesenchymal stem cell sheet,and to analyze the feasibility of preventing esophageal stenosis after esophageal endoscopic submucosal dissection.METHODS:The green fluorescent protein-labeled adipose-derived mesenchymal stem cell sheets were prepared using temperature-sensitive culture dishes and were set as the experimental group.Conventionally adherent adipose-derived mesenchymal stem cells were used as the control group.Levels of vascular endothelial growth factor,transforming growth factorβ1and hepatocyte growth factor in the two groups were compared by ELISA,RT-qPCR and western blot.The skin ulcers of nude mice were prepared,and randomized into cell sheet group and model group.The green fluorescent protein-labeled adipose-derived mesenchymal stem cell sheets were then implanted into the skin wound in the cell sheet group.RESULTS AND CONCLUSION:The number of cells in the cell sheet was high with enriched extracellular matrix,and there was an interconnection between cells.The expression levels of vascular endothelial growth factor,transforming growth factorβ1and hepatocyte growth factor in the experimental group were all higher than those in the control group.The adipose-derived mesenchymal stem cell sheet on the back of nude mice was successfully transplanted,and the wound healing in the cell sheet group was faster than that of the model group.Experimental findings indicate that adipose-derived mesenchymal stem cell transplantation combined with the cell sheet technology is feasible to prevent stenosis after esophageal endoscopic submucosal dissection,but a large number of basic and clinical studies are warranted.