载肝细胞生长因子的壳聚糖基原位水凝胶对角膜内皮损伤的修复作用及机制研究

角膜疾病是全球导致失明的主要原因之一。角膜移植作为治疗角膜盲的有效方法，受到健康角膜供体严重匮乏的制约。具有生物活性的组织工程角膜为该问题提供了新的解决方案。本实验室前期研究表明，羟丙基壳聚糖具有良好的生物相容性，作为角膜内皮细胞载体具有一定的可行性。针对角膜载体材料存在的降解性不合适和机械性能不足的缺点，申请人拟以氧化透明质酸作为生物交联剂，通过调整反应条件和组分构成，与羟丙基壳聚糖和明胶制备利于细胞贴附和增殖，具有改善的降解性和机械性能的可注射原位复合水凝胶。该水凝胶包载HGF后，可进一步优化角膜细胞移植后的微环境，减小炎症反应和角膜纤维化问题。本项目将阐明复合水凝胶与两种来源的角膜内皮细胞间的相互作用，通过建立兔角膜内皮缺损模型，运用多层次方法研究组织工程化角膜内皮植片的修复作用，并探索其修复机制。本项目的实施有望为组织工程角膜的研究提供新的思路，为临床角膜损伤的治疗提供实验依据。

Corneal disease is one of the main causes of blindness worldwide. Corneal transplantation is the effective method to treat corneal blindness, which is restricted by the serious shortage of healthy corneal donors. The tissue engineering cornea with biological activity provides a new solution to this problem. Our previous studies showed that hydroxypropyl chitosan had good biocompatibility and was feasible as corneal endothelial carrier. In view of the shortcomings of improper degradability and insufficient mechanical properties of corneal carrier materials, the oxidized hyaluronic acid is used as a biological crosslinking agent. By adjusting the reaction conditions and components, oxidized hyaluronic acid, hydroxypropyl chitosan and gelatin are used to prepare injectable in-situ composite hydrogel with improved biodegradability and mechanical properties, which is beneficial for cell attachment and proliferation. The hydrogel can further optimize the microenvironment after corneal transplantation by encapsulating HGF, and reduce the inflammatory reaction and corneal fibrosis. This study will elucidate the interaction between composite hydrogel and two kinds of corneal endothelial cells. By establishing the rabbit corneal endothelial defect model, we will study the repair effect of the tissue engineering corneal endothelial graft by multi-level methods, and explore its repair mechanism. The implementation of this study will provide a new idea for the research of tissue engineering cornea and an experimental basis for the treatment of clinical corneal injury.