预构血管化、上皮化组织工程气管修复大动物长段气管缺损及其上皮功能重建机制的研究

长段气管缺损的修复是临床治疗难题。组织工程技术为其提供了新思路，但大动物模型上长段复合气管构建与修复技术一直未获突破。申请人前期成功构建了完全上皮化、血管化并含管状软骨的类气管复合组织，但如何利用该复合组织实现长段气管缺损修复？移植的口腔上皮远期能否实现功能重建？以及如何在术后早期即实现上皮生理功能重建等关键科学问题尚未解决，成为阻碍该技术临床转化的关键。针对上述问题，本项目提出以下解决方案：1）利用该气管复合物结合显微外科带蒂转移技术以及前期气管缺损修复和护理经验，建立长段气管缺损修复大动物模型；2）利用离体活组织绿色荧光蛋白标记技术实现移植口腔上皮长期标记，明确移植后口腔上皮远期转归并阐明其修复机制；3）建立气道上皮细胞膜片构建技术，早期实现仿真气管上皮功能重建。本项目将全面建立组织工程技术修复长段气管缺损的大动物模型，优化上皮重建策略并阐明其修复机制，为临床应用转化提供坚实理论依据。

The repair of long trachea defect is a difficult problem in clinical treatment. Tissue engineering technology provides a new idea for it, but there have been no breakthroughs in the construction of long-segment composite tracheas and the following repair of tracheal defects in large animals. The applicant successfully constructed a epithelialized and vascularized trachea-like composite tissue containing tubular cartilage in previous study. However, there are still issues that remain including how to use the composite tissue to repair the defect of long-segment trachea, what is the long-term outcome of the transplanted oral epithelium transplantation, and how to reconstruct physiological epithelium in the early postoperative stage. All of these have become the key to hinder the clinical transformation of this technology. Due to the above problems, the following solutions are proposed in this project: 1) Using the tracheal complex combined with microsurgical pedicle transfer technology and previous experience of tracheal defect repair to establish a large animal model for the repair of long-segment tracheal defects; 2) Achieving long term fluorescent labeling of oral epithelium after transplantation through green fluorescent protein labeling technology of living tissue in vitro to determine the long-term outcome of transplanted oral epithelium and help elucidate its repair mechanism; 3) Finally, establishing the technology for construction of airway epithelial cell membrane to achieve the physiological simulation of tracheal epithelial function in the early stage. This project will comprehensively establish a large animal model of tissue engineering technology for repairing long tracheal defects, optimize the epithelial reconstruction strategy and clarify their repair mechanisms, so as to provide a solid theoretical basis for clinical application and transformation.