Human Corneal Model for Ocular Irritation Assay

用于眼刺激测定的人类角膜模型

• 批准号: 8004890
• 负责人: Yulia Kaluzhny
• 金额: $ 35.22万
• 依托单位: MATTEK CORPORATION
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8004890
• 关键词: Address; Adherent Culture; Animal Testing; Animal Welfare; Animals; Biological Assay; Cells; Chemical Injury; Chemicals; Classification; Cornea; Corneal Endothelium; Data; Databases; Development; Economics; Endothelial Cells; Epithelial; Epithelial Cells; Epithelium; Evaluation; Exhibits; Exposure to; Eye; Eye Injuries; Generations; Goals; Harvest; Human; In Vitro; Industry; Injury; Irritants; Laboratories; Materials Testing; Measurement; Methods; Modeling; Oryctolagus cuniculus; Pathology; Performance; Pharmacologic Substance; Phase; Phenotype; Physiology; Production; Proliferating; Quality Control; Recovery; Reliance; Reproducibility; Safety; Scientist; Stromal Cells; Testing; Thick; Time; Tissue Model; Tissues; Variant; Wound Healing; base; corneal epithelium; experience; in vitro Assay; in vitro testing; in vivo Model; irritation; research study; response; time use; tool

DESCRIPTION (provided by applicant): Replacement of the Draize rabbit eye test for ocular irritancy testing remains an elusive goal. Many believe a corneal full thickness (CFT) model is necessary to completely model the interaction of exogenous chemicals with human cornea. During Phase 1, human corneal epithelial, stromal, and endothelial cells were isolated from native tissue, expanded in monolayer culture, and cryopreserved. Culture conditions were identified to produce highly differentiated 2-layer (corneal epithelium and keratocyte-containing stroma) and 3- layer (epithelium, stroma, and endothelium) CFT tissues. The CFT tissues exhibited improvements over epithelial tissues in terms of barrier and differentiated phenotype. An assay and prediction model (PM) was developed which allowed the CFT tissue to distinguish between irritants and non-irritants with 100% accuracy. Other functional studies showed that the CFT tissue appears suitable to evaluate time to recovery (TTR) and depth of injury (DOI) following chemical injury. Finally, an analysis of CFT tissue production showed that the economics of CFT are favorable. Phase II will expand upon the Phase 1 result to further develop the CFT tissue model. Initial goals will focus on preparing the CFT tissue model for commercial production - cell stocks will be expanded to insure a stable cell supply and quality control parameters will be developed. The database of materials tested using the CFT will be expanded, the PM for irritant/non-irritant classification will be finalized, and inter-laboratory transferability of the assay will be demonstrated. Finally, the CFT tissues will be used for TTR and DOI measurements and an assay and prediction model for replacement of the Draize test will be developed. Due to these enhanced capabilities, the CFT will provide toxicologists and scientists with a tool that will dramatically decrease, if not entirely eliminate, the need for animal testing to determine ocular irritancy. PUBLIC HEALTH RELEVANCE: Safety assessment of all new chemicals and products requires evaluation of potential ocular irritancy in case of intentional or unintentional exposure to the eye. Current animal-based test methods are suboptimal because of animal welfare concerns and in vitro methods lack the complexity required to completely model in vivo responses. We will develop a corneal full thickness tissue and in vitro test methods that will dramatically reduce or eliminate the need to use animals to assess ocular irritancy of chemicals and products.

描述（由申请人提供）：取代用于眼刺激性测试的 Draize 兔眼测试仍然是一个难以实现的目标。许多人认为角膜全层（CFT）模型对于完整模拟外源化学物质与人类角膜的相互作用是必要的。在第一阶段，从天然组织中分离人角膜上皮细胞、基质细胞和内皮细胞，在单层培养物中扩增并冷冻保存。确定培养条件可产生高度分化的 2 层（角膜上皮和含有角膜细胞的基质）和 3 层（上皮、基质和内皮）CFT 组织。 CFT 组织在屏障和分化表型方面比上皮组织表现出改善。开发了一种检测和预测模型 (PM)，使 CFT 组织能够以 100% 的准确度区分刺激物和非刺激物。其他功能研究表明，CFT 组织似乎适合评估化学损伤后的恢复时间 (TTR) 和损伤深度 (DOI)。最后，对 CFT 卫生纸生产的分析表明，CFT 的经济性是有利的。第二阶段将扩展第一阶段的结果，进一步开发 CFT 组织模型。最初的目标将集中于准备用于商业生产的 CFT 组织模型 - 将扩大细胞库存以确保稳定的细胞供应，并制定质量控制参数。使用 CFT 测试的材料数据库将得到扩展，刺激性/非刺激性分类的 PM 将最终确定，并且将证明检测的实验室间可转移性。最后，CFT 组织将用于 TTR 和 DOI 测量，并将开发替代 Draize 测试的测定和预测模型。由于这些增强的功能，CFT 将为毒理学家和科学家提供一种工具，即使不能完全消除，也将大大减少通过动物测试来确定眼部刺激性的需要。 公共健康相关性：对所有新化学品和产品的安全评估需要评估有意或无意接触眼睛时潜在的眼部刺激性。由于动物福利问题，目前基于动物的测试方法并不理想，而且体外方法缺乏完全模拟体内反应所需的复杂性。我们将开发角膜全层组织和体外测试方法，这将大大减少或消除使用动物来评估化学品和产品对眼部刺激性的需要。