The purpose of the current work was to utilize a three dimensional (3D) corneal epithelial tissue model to study dry eye disease and oxidative stress-related corneal epithelial injuries for the advancement of ocular therapeutics. Air-liquid interface cultures of normal human corneal epithelial cells were used to produce 3D corneal epithelial tissues appropriate for physiologically relevant exposure to environmental factors. Oxidative stress was generated by exposing the tissues to non-toxic doses of ultraviolet radiation (UV), hydrogen peroxide, vesicating agent nitrogen mustard, or desiccating conditions that stimulated morphological, cellular, and molecular changes relevant to dry eye disease. Corneal specific responses, including barrier function, tissue viability, reactive oxygen species (ROS) accumulation, lipid peroxidation, cytokine release, histology, and gene expression were evaluated. 3D corneal epithelial tissue model structurally and functionally reproduced key features of molecular responses of various types of oxidative stress-induced ocular damage. The most pronounced effects for different treatments were: UV irradiation - intracellular ROS accumulation; hydrogen peroxide exposure - barrier impairment and IL-8 release; nitrogen mustard exposure - lipid peroxidation and IL-8 release; desiccating conditions - tissue thinning, a decline in mucin expression, increased lipid peroxidation and IL-8 release. Utilizing a PCR gene array, we compared the effects of corneal epithelial damage on the expression of 84 oxidative stress-responsive genes and found specific molecular responses for each type of damage. The topical application of lubricant eye drops improved tissue morphology while decreasing lipid peroxidation and IL-8 release from tissues incubated at desiccating conditions. This model is anticipated to be a valuable tool to study molecular mechanisms of corneal epithelial damage and aid in the development of therapies against dry eye disease, oxidative stress- and vesicant-induced ocular injuries.

中文翻译：

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不同来源的角膜损伤中的氧化应激：3D人角膜上皮组织模型的利用。

当前工作的目的是利用三维（3D）角膜上皮组织模型来研究干眼病和与氧化应激相关的角膜上皮损伤，以促进眼科治疗的发展。正常人角膜上皮细胞的气液界面培养用于产生3D角膜上皮组织，适合于生理学上暴露于环境因素。氧化应激是通过将组织暴露于无毒剂量的紫外线（UV），过氧化氢，发泡剂氮芥末或刺激与干眼病相关的形态，细胞和分子变化的干燥条件而产生的。角膜特异性反应，包括屏障功能，组织活力，活性氧（ROS）积累，脂质过氧化，细胞因子释放，组织学和基因表达进行了评估。3D角膜上皮组织模型在结构和功能上重现了各种类型的氧化应激诱导的眼损伤的分子反应的关键特征。不同治疗方法最明显的作用是：紫外线照射-细胞内ROS的积累；过氧化氢暴露-屏障障碍和IL-8释放; 氮芥暴露-脂质过氧化和IL-8释放; 干燥条件-组织变薄，粘蛋白表达下降，脂质过氧化增加和IL-8释放。利用PCR基因阵列，我们比较了角膜上皮损伤对84种氧化应激反应基因表达的影响，并发现了每种损伤类型的特异性分子反应。润滑剂滴眼液的局部应用改善了组织形态，同时减少了在干燥条件下培养的组织中的脂质过氧化作用和IL-8释放。预期该模型将是研究角膜上皮损伤的分子机制并帮助开发抗干眼病，氧化应激和药物引起的眼外伤的疗法的有价值的工具。