承载人脐带间充质干细胞的水凝胶体系修复食管粘膜损伤及防治食管良性狭窄的应用研究

A large area of esophageal mucosal injury caused by inflammation, drugs and surgical corrosions often complicate benign stricture. The treatment is extremely difficult. The effective reparation of the esophageal mucosa, prevention and treatment of benign stenosis are serious problems. Human umbilical cord mesenchymal stem cells (hUCMSCs) are potential induced to differentiate into the epithelial cells. Its development of tissue engineering has provided some new ideas. This study is to accomplish the following tasks: (1) isolate, culture and identify hUCMSCs, recombined with the hydrogel thermally induced by self-developed PEG / biodegradable polyester block copolymers, construct hUCMSCs-carried hydrogel and hydrogel patch; (2) verify the biocompatibility of hydrogel as a carrier of hUCMSCs and explore the mechanisms of hUCMSCs settling in the lesion place, differentiating toward endothelial cells and repairing the injured mucosal epithelial differentiation mechanism to cells and repair by using in vitro and in vivo rat model of esophageal injury transplantation; (3) endoscopicly construct esophageal mucosal injury model and total-circumference mucosal defect model of big animals to evaluated the function of hUCMSCs-carried hydrogel and hydrogel patch transplanting and adhereing to the injury place and accelerating the wound regeneration, preventing scarring, prevention and treatment of esophageal stenosis; (4) provide theoretical and experimental evidences for the repairation and regeneration of esophageal mucosa, prevention and treatment of benign esophageal stricture..

炎症、药物腐蚀及手术等原因造成的大面积食管粘膜损伤（缺损）常并发良性狭窄，治疗极为棘手。如何有效修复食管粘膜，预防和治疗良性狭窄，是亟待解决的问题。人脐带间充质干细胞（hUCMSCs）具有诱导分化为上皮细胞的潜能，其在组织工程学的发展，为我们提供了新的思路。该课题将分离培养鉴定hUCMSCs，与自主研发的PEG/可降解聚酯嵌段共聚物热致水凝胶复合，构建承载hUCMSCs的水凝胶及水凝胶补片。通过体外诱导和大鼠食管损伤模型体内移植，验证水凝胶作为hUCMSCs载体的生物相容性，探讨hUCMSCs在病变部位定居、向上皮细胞分化能力及修复受损粘膜的机制。内镜下构建食管粘膜损伤和全周粘膜缺损大动物模型，分别评价承载hUCMSCs的水凝胶及补片，移植或贴覆在损伤部位，加快创面愈合，防止瘢痕形成的价值。为修复、再生食管粘膜，预防和治疗食管良性狭窄，提供理论基础和实验依据。

本课题针对炎症、药物腐蚀及内镜粘膜下剥离术（ESD）等原因造成的大面积食管粘膜损伤（缺损）并发的良性狭窄问题进行研究。根据课题申请中的研究内容实施，主要分为五部分。第一部分为分离培养鉴定人脐带间充质干细胞并荧光标记；第二部分为构建承载hUCMSCs的水凝胶体系；第三部分为构建大鼠食管损伤模型，研究承载hUCMSCs的水凝胶体系的作用；第四部分为构建猪食管损伤模型，研究承载hUCMSCs的水凝胶体系的作用；第五部分为构建猪ESD术后食管全周粘膜缺损模型，探讨承载hUCMSCs的水凝胶体系的应用价值。.在课题研究期间：（1）顺利完成人脐带间充质干细胞培养及鉴定，并对人脐带间充质干细胞体外分化能力进行鉴定。（2）成功构建承载hUCMSCs的水凝胶体系，在前期试验基础上，测试完成新型水凝胶体系的生物安全性，项目写入“生物医用材料研发与组织器官修复替代”国家重点研发计划项目“环境响应医用水凝胶”指南。已和威高集团达成合作意向，进入实质性合作阶段，并发表SCI论文1篇。（3）顺利建立大鼠和大动物（猪）食管狭窄模型。（4）确定采用金属夹直接固定法、补片+金属全覆膜支架法、微小补片（浆液）+金属全覆膜支架法以及局部原位皮瓣移植法四种治疗方法可用于食管良性狭窄的预防和治疗，首次报道食道皮瓣移植技术，首次报道良性狭窄段气囊切割技术，救治成功1例食管、胃全消化道烧伤后狭窄的患者。.目前共完成SCI论文4篇，发表2篇。申请发明专利4项，授权2项。培养研究生2名。