Cell and Molecular Biology Laboratory, Department of Ophthalmology, University Hospital, LMU Munich, Munich, Germany.
Institute of Human Anatomy and Embryology, University of Regensburg, Regensburg, Germany.
Acta Ophthalmol. 2020 Sep;98(6):e773-e780. doi: 10.1111/aos.14366. Epub 2020 Feb 3.
To determine whether excimer laser ablation of guttae is a viable strategy for removal of diseased tissue in Fuchs' endothelial corneal dystrophy (FECD) on excised human Descemet membranes and whether an excimer laser-created wound on healthy human corneas ex vivo is recolonized with corneal endothelial cells.
Descemet membranes of FECD patients and corneal endothelium of normal human corneas were ablated ex vivo using an excimer laser licensed for glaucoma surgery. Specimens were kept in cell culture medium supplemented with 10 μm of rho-kinase inhibitor ripasudil. Corneal endothelial cell regeneration was observed using light and electron scanning microscopy. Furthermore, the whole corneal samples were evaluated by haematoxylin/eosin staining and immunohistochemical analysis using antibodies against Na /K -ATPase.
Guttae and corneal endothelium could be ablated with an excimer laser without total ultrastructural damage to the Descemet membrane or stroma. Nearly complete endothelial wound closure was accomplished after 26-38 days in treated corneas. Light and electron scanning microscopy suggested the establishment of a layer of flat endothelial cells. Additionally, Na /K -ATPase expression could only be observed on the inner side of the Descemet membrane.
Our proof of concept study demonstrated that excimer lasers can be used to ablate diseased tissue from excised FECD Descemet membranes ex vivo. Additionally, corneal endothelial cells recolonize a previously ablated endothelial area in healthy human corneas ex vivo under treatment with ripasudil. Thus, our results are the first experimental basis to further investigate the feasibility of an excimer laser ablation as a graftless FECD treatment option.
确定准分子激光消融对于切除 Fuchs 内皮角膜营养不良（FECD）患者的病变组织是否可行，以及准分子激光在健康人角膜上造成的伤口是否会被角膜内皮细胞重新占据。
使用已获得青光眼手术许可的准分子激光，对 FECD 患者的后弹力膜和正常人的角膜内皮进行离体消融。标本保存在添加了 Rho 激酶抑制剂 ripasudil（10μm）的细胞培养基中。使用光镜和电子扫描显微镜观察角膜内皮细胞的再生情况。此外，对整个角膜样本进行苏木精/伊红染色和免疫组织化学分析，使用针对 Na / K -ATP 酶的抗体进行评估。
准分子激光可以消融角膜内皮和后弹力层，而不会对后弹力膜或基质造成完全的超微结构损伤。在治疗后的角膜中，经过 26-38 天，几乎可以完全闭合内皮伤口。光镜和电子扫描显微镜显示，建立了一层平坦的内皮细胞层。此外，Na / K -ATP 酶的表达仅能在前弹力膜的内侧观察到。
我们的概念验证研究表明，准分子激光可用于切除离体 FECD 后弹力膜上的病变组织。此外，在 ripasudil 治疗下，健康人角膜的先前消融的内皮区域可以被角膜内皮细胞重新占据。因此，我们的研究结果为进一步研究准分子激光消融作为无移植物 FECD 治疗选择的可行性提供了首个实验依据。
