Validation of a Human In Vitro Vaginal Irritation Test

描述（由申请方提供）：在I期研究期间，研究了6种模型试验材料对高度分化的阴道-宫颈外（VEC）组织模型的影响。监测对组织活力、结构、屏障功能和炎性细胞因子释放的影响。许多试验得出的结果相互支持，增加了体外结果的可信度。例如，组织损伤和细胞因子释放导致组织活力丧失。此外，尽管数据表明屏障功能是更敏感的终点（即，屏障功能降低发生在不降低组织活力的较低浓度下），但当组织活力降低时，测量到屏障功能降低。重要的是，在家兔阴道刺激试验中具有轻度刺激性的一种材料诱导组织活力和屏障功能显著降低，并增加炎症介质释放。此外，经济分析表明，体外方法是目前使用的家兔阴道刺激试验方法的一种具有成本效益的替代方法。在第2阶段，将进一步扩大体外试验系统的效用，以预测长期重复暴露后的阴道刺激。使用第1阶段开发的终点，将开发预测模型，以准确评估供试品和制剂的体内阴道刺激。该方法将转移至外部合同检测实验室，并将进行多实验室GLP验证研究。此外，还将评估试验的短期和长期重现性，并开发高通量版本的试验方法。公共卫生相关性：用于评估化学品和制剂的阴道刺激潜力的预测性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。评估阴道刺激对于减少数百万妇女的化学危害非常重要。所提出的基于人类重建组织的系统将提供一种灵敏且经验证的测定方法，用于筛选具有阴道刺激潜力的化学品/制剂。此外，该测定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.