Human Corneal Model for Ocular Irritation Assay

描述(由申请人提供)：替代Draize兔眼测试用于眼刺激测试仍然是一个难以实现的目标。虽然单层培养和分层角膜上皮组织已经被开发出来，并可用作眼刺激的初始筛选，但它们不能完全模拟外源化学物质与人类角膜组织的相互作用。例如，用目前的组织模型开发的方案正确识别了高比例的眼睛刺激物(~95%)，但错误地将很大比例的非刺激物归类为刺激物(~33%)。此外，这些模型也不能模拟化学伤害后的损伤程度和恢复时间。因此，仍然需要进行动物或人体临床试验，以获得有关化学物质暴露于眼睛的影响的竞争性图景。目前的应用将开发一种折衷内皮、基质和上皮成分的角膜全厚度(CFT)组织模型。CFT组织将含有人类细胞，并产生与天然角膜组织相当的组织屏障特性。由于其改进的屏障，CFT将避免非刺激性的错误分类。此外，CFT组织将有助于确定有毒化学物质暴露后的损伤程度和恢复时间。由于这些增强的能力，CFT将为毒理学家和科学家提供一种工具，即使不是完全消除，也将极大地减少确定眼睛刺激性的动物测试的需要。与公共健康相关：所有新化学品和产品的安全性评估需要评估有意或无意接触眼睛的情况下潜在的眼睛刺激性。由于对动物福利的担忧，目前基于动物的测试方法是超级最佳的，而体外方法缺乏完全模拟体内反应所需的复杂性。我们将开发一种角膜全厚度组织和体外测试方法，大大减少或消除使用动物来评估化学品和产品对眼睛的刺激性的需要。

DESCRIPTION (provided by applicant): Replacement of the Draize rabbit eye test for ocular irritancy testing remains an elusive goal. Although monolayer cultures and stratified corneal epithelial tissues have been developed and are useful as initial screens for ocular irritancy, they are not capable of completely modeling the interaction of exogenous chemical with human corneal tissue. For instance, protocols developed with current tissue models correctly identify a high percentage of ocular irritants (~ 95%) but misclassify a large percentage of non-irritants as irritants (~33%). Also, these models are not capable of modeling extent of injury and time to recovery following chemical insult. Thus, animal or human clinical tests are still needed to gain a compete picture of the effects of chemical exposure to the eye. The current application will develop a corneal full thickness (CFT) tissue model compromised of endothelial, stromal, and epithelial components. The CFT tissue will contain human cells and develop tissue barrier properties that are comparable to native corneal tissue. Because of its improved barrier, the CFT will avoid the misclassification of non-irritants. In addition, the CFT tissue will be useful in determining the extent of injury and time to recovery following noxious chemical exposure. Due to these enhanced capabilities, the CFT will provide toxicologists and scientists with a tool that will dramatically decrease, if not eliminate entirely, 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 sup-optimal 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.