飞秒激光胶原交联效应的研究

Corneal ectasic diseases include primary and secondary keratoconus that dramatically damage visual acuity, and become one of the long-term risks after refractive surgeries. Collagen cross-linking (CXL) with a femtosecond laser will improve the stability of corneal biomechanics, but the infiltration effect and the mechanism of corneal wound healing postoperatively remain unclear. This project intends to evaluate the rational energy/time ratio of the operation, the characteristics of keratocyte apoptosis and nerve recovery postoperatively. Based on our previous studies, it is planned to observe corneal morphological changes after the CXL with a femtosecond laser and traditional CXL using the confocal microscope. We will make a comparison of keratocyte apoptosis, proliferation and inflammation postoperatively by an immunohistochemical technique for TUNEL and the expression of Ki-67 and CD11b. Also, the corneal nerve injury and biomechanics properties changes will be taken into evaluation. The purpose of this project is to provide the theoretical basis for the treatment of corneal ectasic diseases.

扩张性角膜病变包括原发或继发的圆锥角膜严重损害视力，亦为屈光手术远期风险之一。飞秒激光辅助角膜胶原交联（collagen cross-linking, CXL）有望改善角膜生物力学稳定性，但其渗透效应以及创伤愈合规律有待阐述。本项目旨在探讨飞秒激光胶原交联的能量/时间比的优化规律、细胞凋亡及神经修复特点。拟在申请者前期研究基础上，通过活体共焦显微镜观察飞秒激光CXL和常规CXL术后角膜组织的形态结构改变，采用免疫组化分析术后角膜TUNEL反应并检测ki-67及CD11b的表达，进而比较两种手术后角膜基质细胞凋亡、增殖及炎性反应的差异，并对术后角膜神经损伤及生物力学性能的变化进行评估。该研究将为临床扩张性角膜疾病的干预提供理论依据。

紫外光-核黄素胶原交联术后组织变化。本研究项目的结果包括两方面内容：（1）观察飞秒辅助交联对角膜形态学的影响；（2）探讨促渗剂辅助角膜胶原交联术后角膜愈合规律。.在第一方面，先期动物实验的摸索，发现了常规交联和飞秒交联术后角膜生物力学特性的变化特点，以及术后早期角膜地形图的变化规律，并为后期分子生物学方面的实验进行了预实验尝试的研究取得了重要成果。.在第二方面，研究5.4J和7.2J高低不同能量组促渗剂辅助角膜胶原交联，术后1月内裂隙灯显微镜的观察情况，观察光镜和透射电子显微镜下角膜形态结构变化特点，并采用免疫组织化学方法检测Ki-67及CD11b的表达结果，进一步研究了跨上皮交联是否引起的角膜增殖炎症更轻的问题。