Validation of a Human In Vitro Vaginal Irritation Test

描述（由申请人提供）：在第一阶段的研究中，研究了6种模型试验材料对高度分化阴道-宫颈外（VEC）组织模型的影响。监测组织活力、结构、屏障功能和炎症细胞因子释放的影响。许多测定的结果相互支持，增加了对体外结果的信心。例如，组织损伤和细胞因子释放平行于组织活力的丧失。此外，尽管数据表明屏障功能是一个更敏感的终点（即在较低浓度下发生屏障功能下降，但不会降低组织活力），但当组织活力下降时，屏障功能也会下降。重要的是，在兔阴道刺激试验中，轻度刺激的一种材料导致组织活力和屏障功能显著降低，炎症介质释放增加。此外，经济分析表明，体外方法是一种具有成本效益的替代目前使用的兔阴道刺激试验方法。在第二阶段，体外检测系统的应用将进一步扩大，以预测慢性、重复暴露后的阴道刺激。使用第一阶段开发的终点，将开发一个预测模型来准确评估测试材料和配方的体内阴道刺激。该方法将转移到外部合同测试实验室，并进行多实验室GLP验证研究。此外，将评估该分析的短期和长期重复性，并将开发该分析方法的高通量版本。公共卫生相关性：用于评估化学品和配方的阴道刺激潜力的预测测试系统将在涉及妇女护理产品，杀微生物剂，避孕药，赋形剂开发和局部药物产品的行业中具有深远的应用。评估阴道刺激对减少化学品对数百万妇女的危害很重要。拟建的基于人体重建组织的系统将为筛选具有阴道刺激潜力的化学品/配方提供一种敏感和有效的分析方法。此外，该分析方法将具有成本效益，并减少实验动物的使用。

DESCRIPTION (provided by applicant): During Phase 1 research, the effects of 6 model test materials on a highly differentiated vaginal- ectocervical (VEC) tissue model were studied. Effects on tissue viability, structure, barrier function, and inflammatory cytokine release were monitored. Many of the assays gave results which supported one another and increased confidence in the in vitro results. For instance, histological damage and cytokine release paralleled losses in tissue viability. In addition, decreases in barrier function were measured when tissue viability decreased although the data indicated that barrier function was a more sensitive endpoint (i.e. decreases in barrier function occurred at lower concentrations that did not decrease tissue viability). Importantly, the one material which was mildly irritating in the rabbit vaginal irritation test induced significant decreases in tissue viability and barrier function and increased inflammatory mediator release. In addition, an economic analysis showed the in vitro method to be a cost-effective alternative to the currently used rabbit vaginal irritation test methodology. During Phase 2, the utility of the in vitro assay system will be further expanded to predict vaginal irritation following chronic, repeat exposure. Using the endpoints developed in Phase 1, a prediction model will be developed to accurately assess the in vivo vaginal irritation of test materials and formulations. The method will be transferred to outside contract testing labs and a multi-lab GLP validation study will be performed. In addition, the short and long term reproducibility of the assay will be assessed, and a high throughput version of the assay method will be developed. PUBLIC HEALTH RELEVANCE: A predictive test system for assessing the vaginal irritation potential of chemicals and formulations will have far reaching application in industries involved in women's care products, microbicide, contraceptives, excipient development, and topical pharmaceutical products. Evaluation of vaginal irritation is important to minimize chemical hazards to millions of women. The proposed human reconstructed tissue based system will provide a sensitive and validated assay method for screening of chemicals/formulations with vaginal irritation potential. Furthermore, the assay method will be cost effective and reduce the use of laboratory animals for experimentation.