ATotally Animal-free Model for Acute Chemicl Toxicity Testing

用于急性化学毒性测试的完全无动物模型

• 批准号: 8787470
• 负责人: SHAY-WHEY M KOH
• 金额: $ 22.56万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8787470
• 关键词: Acute; Address; Agreement; Algorithms; Animals; Automation; Binding; Categories; Cattle; Cell Culture Techniques; Cell surface; Cells; Chemicals; Chickens; Computers; Conscious; Consensus; Cornea; Corneal Endothelium; Corneal Opacity; Corrosives; Cosmetics; Data; Endothelial Cells; Environment; Evaluation; Extravasation; Eye; Eye Injuries; Fluorescein; Goals; Health; Hour; Household; Household Products; Human; Human Resources; Image Analysis; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Iris; Irritants; Keratoplasty; Life; Measures; Methods; Modeling; Morals; One-Step dentin bonding system; Oryctolagus cuniculus; Outcome; Permeability; Play; Process; Property; Reaction; Role; Scientist; Self Care; Serum; Staining method; Stains; Surgeon; System; Techniques; Testing; Toxicity Tests; Transplantation; Work; base; conjunctiva; corneal epithelium; hazard; in vitro Model; in vivo; injured; insight; irritation; method development; safety testing

DESCRIPTION (provided by applicant): The Draize eye irritation test has been in use since 1944. One eye of a conscious rabbit is instilled with the test substance, while the other eye is the control. The reactions of the cornea, conjunctiva, and iris are then evaluated in a period of one hour to three weeks. Because of the test's moral and scientific short comings, a replacement has been actively sought, but there is still none in the horizon. Today, the Draize test remains the only one that can classify a substance into one of the four categories of eye irritation: corrosive/severe, moderate, mild, and non- irritants. Specifically, none of the in viro methods can identify the moderate and mild irritants and the Draize test continues. None of the in vitro method addresses the issue of ocular inflammation, an integral part of the Draize eye irritation test brought upon by dying cells following test substance instillation. We hypothesize that an in vitro method with an end point that can predict the inflammatory potential of the injured eye can identify moderate and mild irritants, in addition to the corrosive/severe and non- irritants. Using human donor corneas not used for transplantation, our preliminary studies demonstrated in a totally animal (and animal product)-free system that the levels of corneal endothelial damage, revealed by alizarin red S staining and quantified by image analysis of the red stain in the panoramic view of the corneal endothelium, predicted with 100% accuracy the irritation potential of 18 test substances in the corrosive/severe, moderate, mild, and non- irritants categories. Our goal is to demonstrate that our method can fully replace the Draize eye irritation test. The specific aims are to 1) use our method to classify test substances that have historical Draize test data available in public, 2) study the status of VIP in the damaged corneal endothelium in the test substance-treated human donor corneas.

描述(申请人提供)：自1944年开始使用Draize眼刺激试验。清醒的兔子的一只眼睛被注入测试物质，而另一只眼睛是对照。然后在一小时到三周的时间内评估角膜、结膜和虹膜的反应。由于这项测试在道德和科学上的缺陷，人们一直在积极寻找替代方案，但目前还没有出现。今天，Draze测试仍然是唯一一种可以将一种物质归类为四种眼睛刺激之一的测试：腐蚀性/重度、中度、轻度和非刺激性。具体地说，没有一种病毒内方法可以识别中度和轻度刺激物，Draize测试继续进行。没有一种体外方法解决眼部炎症的问题，眼部炎症是Draize眼刺激试验的一个组成部分，由试验物质滴注后死亡的细胞引起。我们假设，一种具有终点的体外方法可以预测受伤眼睛的炎症潜在性，除了腐蚀性/重度和非刺激性之外，还可以识别中度和轻度刺激性。使用未用于移植的人供体角膜，我们的初步研究表明，在完全不含动物(和动物产品)的系统中，茜素红S染色显示的角膜内皮损伤水平，以及角膜内皮全景红色染色的图像分析，可以100%准确地预测腐蚀性/重度、中度、轻度和非刺激性类别的18种测试物质的刺激潜力。我们的目标是证明我们的方法可以完全取代Draize眼刺激测试。其具体目的是：1)使用我们的方法对已公开的历史Draize测试数据的测试物质进行分类；2)研究经测试物质处理的人供体角膜中VIP在受损角膜内皮中的状态。