Limbal stem cell deficiency (LSCD) secondary to ocular surface alkali burn is a blinding condition that features corneal conjunctivalization. Mechanistic insights into its pathophysiology are lacking. Here, we developed a mouse model that recapitulates human disease to comprehensively delineate the clinicopathological features of a conjunctivalized cornea. LSCD was induced in the right eyes of 6-8-week-old C57BL/6 male and female mice () by topical administration of 0.25N sodium hydroxide on the cornea. Uninjured left eyes served as controls. Clinical, histological, phenotypic, molecular, and immunological assessments were performed at multiple time-points over 6-months. Clinically, alkali burn caused persistent corneal opacity (p = 0.0014), increased punctate staining (p = 0.0002), and reduced epithelial thickness (p = 0.0082) compared to controls. Total LSCD was confirmed in corneal whole mounts by loss of K12 protein (p < 0.0001) and mRNA expression (p = 0.0090). Instead, K8, K13, K15 and MUC5AC conjunctival epithelia prevailed. 20 % of injured corneas developed islands of K12 epithelia, suggesting epithelial transdifferentiation. Squamous metaplasia was detected in 50 % of injured corneas. Goblet cell density peaked early post-injury but decreased over time (p = 0.0047). Intraepithelial corneal basal nerve density remained reduced even at 6-months post-injury (p = 0.0487). We developed and comprehensively characterized a preclinical mouse model of alkali-induced LSCD. Understanding the pathophysiological processes that transpire on the ocular surface in LSCD is key to discovering, testing, and advancing biological and pharmacological interventions that can be dispensed prior to or in conjunction with stem cell therapy to rehabilitate the cornea and restore vision.

中文翻译：

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碱诱导角膜缘干细胞缺陷的临床前小鼠模型的开发和表征


继发于眼表碱烧伤的角膜缘干细胞缺乏症（LSCD）是一种以角膜结膜化为特征的致盲性疾病。缺乏对其病理生理学的机制见解。在这里，我们开发了一种重现人类疾病的小鼠模型，以全面描述结膜角膜的临床病理特征。通过在角膜上局部施用 0.25N 氢氧化钠，在 6-8 周大的 C57BL/6 雄性和雌性小鼠的右眼中诱导 LSCD。未受伤的左眼作为对照。在 6 个月的多个时间点进行了临床、组织学、表型、分子和免疫学评估。临床上，与对照组相比，碱烧伤导致持续性角膜混浊（p = 0.0014）、点状染色增加（p = 0.0002）和上皮厚度减少（p = 0.0082）。通过 K12 蛋白 (p < 0.0001) 和 mRNA 表达 (p = 0.0090) 的损失，在角膜整体上证实了总 LSCD。相反，K8、K13、K15 和 MUC5AC 结膜上皮占主导地位。 20% 受伤的角膜出现 K12 上皮岛，表明上皮转分化。在 50% 受伤的角膜中检测到鳞状上皮化生。杯状细胞密度在损伤后早期达到峰值，但随着时间的推移而下降（p = 0.0047）。即使在受伤后 6 个月，上皮内角膜基底神经密度仍然降低 (p = 0.0487)。我们开发并全面表征了碱诱导 LSCD 的临床前小鼠模型。了解 LSCD 眼表的病理生理过程是发现、测试和推进生物和药物干预措施的关键，这些干预措施可以在干细胞治疗之前或与干细胞治疗结合使用，以修复角膜和恢复视力。