Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine.
Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine;
J Vis Exp. 2023 Jun 30(196). doi: 10.3791/65289.
Corneal neovascularization (CoNV), a pathological form of angiogenesis, involves the growth of blood and lymph vessels into the avascular cornea from the limbus and adversely affects transparency and vision. Alkali burn is one of the most common forms of ocular trauma that leads to CoNV. In this protocol, CoNV is experimentally induced using sodium hydroxide solution in a controlled manner to ensure reproducibility. The alkali burn model is useful for understanding the pathology of CoNV and can be extended to study angiogenesis in general because of the avascularity, transparency, and accessibility of the cornea. In this work, CoNV was analyzed by direct examination under a dissecting microscope and by immunostaining flat-mount corneas using anti-CD31 mAb. Lymphangiogenesis was detected on flat-mount corneas by immunostaining using anti-LYVE-1 mAb. Corneal edema was visualized and quantified using optical coherence tomography (OCT). In summary, this model will help to advance existing neovascularization assays and discover new treatment strategies for pathologic ocular and extraocular angiogenesis.
角膜新生血管化(CoNV)是一种病理性血管生成形式,涉及从角膜缘向无血管角膜生长的血液和淋巴管,并且会对透明度和视力产生不利影响。碱烧伤是导致 CoNV 的最常见眼部创伤形式之一。在本方案中,使用氢氧化钠溶液以可控的方式来实验性诱导 CoNV,以确保重现性。由于角膜的无血管性、透明性和可及性,碱烧伤模型有助于理解 CoNV 的病理学,并且可以扩展到一般的血管生成研究。在这项工作中,通过解剖显微镜直接检查和使用抗 CD31 mAb 对 flat-mount 角膜进行免疫染色来分析 CoNV。通过使用抗 LYVE-1 mAb 对 flat-mount 角膜进行免疫染色来检测淋巴管生成。使用光学相干断层扫描(OCT)可视化和量化角膜水肿。总之,该模型将有助于推进现有的血管生成测定,并发现病理性眼部和眼外血管生成的新治疗策略。