Department of Ophthalmology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, China.
College of Optometry, Wenzhou Medical University, Wenzhou, China; and.
Cornea. 2023 Jul 1;42(7):874-887. doi: 10.1097/ICO.0000000000003242. Epub 2023 Feb 14.
The aim of this study was to develop a rat model of limbal stem cell deficiency (LSCD) by forcing eye-open at birth (FEOB).
A total of 200 Sprague-Dawley neonatal rats were randomly divided into the control group and the experimental group, which received eyelid open surgery on postnatal day 1 (P1). Observation time points were defined as P1, P5, P10, P15, and P30. Slit-lamp microscope and corneal confocal microscope were used to observe the clinical features of the model. The eyeballs were collected for hematoxylin and eosin staining and periodic acid-Schiff staining. Proliferating cell nuclear antigen, CD68/polymorphonuclear leukocytes, and cytokeratin 10/12/13 immunostaining were performed, while the ultrastructure of the cornea was observed by scanning electron microscopy. Real-time polymerase chain reactions (PCRs), western blot, and immunohistochemical staining of activin A receptor-like kinase-1/5 were used to analyze the possible pathogenesis.
FEOB could successfully induce the typical manifestations of LSCD, including corneal neovascularization, severe inflammation, and corneal opacity. In the FEOB group, goblet cells could be detected in the corneal epithelium by periodic acid-Schiff staining. The expression of cytokeratins was also different between the 2 groups. Furthermore, proliferating cell nuclear antigen immunohistochemical staining revealed the weak proliferation and differentiation ability of limbal epithelial stem cells in the FEOB group. Real-time PCRs, western blot, and immunohistochemical staining of activin A receptor-like kinase-1/activin A receptor-like kinase-5 in the FEOB group showed different expression patterns than those of the control group.
FEOB in rats induces ocular surface changes resembling LSCD in humans, representing a novel model of LSCD.
本研究旨在通过生后即刻行眼睑开放术（FEOB）建立大鼠角膜缘干细胞缺乏（LSCD）模型。
200 只 Sprague-Dawley 新生大鼠随机分为对照组和实验组，实验组于生后第 1 天（P1）行眼睑开放术。观察时间点为 P1、P5、P10、P15 和 P30。采用裂隙灯显微镜和角膜共聚焦显微镜观察模型的临床特征。采集眼球行苏木精-伊红染色和过碘酸雪夫染色，行增殖细胞核抗原、CD68/多形核白细胞、细胞角蛋白 10/12/13 免疫组化染色，扫描电镜观察角膜超微结构。采用实时聚合酶链反应（PCR）、Western blot 和免疫组化染色检测激活素 A 受体样激酶-1/5 的表达，分析可能的发病机制。
FEOB 可成功诱导 LSCD 的典型表现，包括角膜新生血管、严重炎症和角膜混浊。在 FEOB 组，过碘酸雪夫染色可检测到角膜上皮中的杯状细胞。2 组间细胞角蛋白的表达也存在差异。此外，增殖细胞核抗原免疫组化染色显示 FEOB 组角膜缘上皮干细胞的增殖和分化能力较弱。FEOB 组的实时 PCR、Western blot 和免疫组化染色显示激活素 A 受体样激酶-1/激活素 A 受体样激酶-5 的表达模式与对照组不同。
FEOB 诱导的大鼠眼表变化类似于人类 LSCD，为 LSCD 提供了一种新的模型。
