ATotally Animal-free Model for Acute Chemicl Toxicity Testing

描述（由申请方提供）：Draize眼刺激试验自1944年以来一直在使用。 清醒家兔的一只眼睛滴注供试品，另一只眼睛为对照。 然后在一小时至三周的时间内评估角膜、结膜和虹膜的反应。 由于该测试的道德和科学的短暂性，人们一直在积极寻找替代品，但仍然没有出现。 今天，Draize测试仍然是唯一一个可以将物质分为四种眼睛刺激类别之一的测试：腐蚀性/严重，中度，轻度和非刺激性。 具体而言，没有一种体外方法可以识别中度和轻度刺激物，Draize试验继续进行。 体外方法均未解决眼部炎症问题，这是Draize眼刺激试验的组成部分，由滴注供试品后死亡细胞引起。 我们假设，具有可以预测受伤眼睛的炎症潜力的终点的体外方法可以识别中度和轻度刺激物，以及腐蚀性/重度和非刺激物。 使用未用于移植的人类供体角膜，我们的初步研究表明，在一个完全动物（和动物产品）-自由系统，角膜内皮损伤的水平，通过茜素红S染色显示，并通过角膜内皮全景图中红色染色的图像分析定量，以100%的准确度预测了18种测试物质在腐蚀性/重度，中度，温和及无刺激性类别。 我们的目标是证明我们的方法可以完全取代Draize眼刺激试验。 具体目的是：1）使用我们的方法对具有公开历史Draize试验数据的供试品进行分类，2）研究供试品处理的人供体角膜中受损角膜内皮中VIP的状态。

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.