Validation of a Human In Vitro Vaginal Irritation Test

描述(由申请人提供)：在第一阶段研究中，研究了6种模型试验材料对高度分化的阴道-外宫颈(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。许多检测结果相互支持，并增加了对体外结果的信心。例如，组织损伤和细胞因子的释放与组织活力的损失是平行的。此外，尽管数据表明屏障功能是一个更敏感的终点(即屏障功能的下降发生在较低浓度，不会降低组织活力)，但当组织活力下降时，屏障功能的下降被测量到。重要的是，在兔阴道刺激试验中有轻微刺激性的一种材料导致组织活力和屏障功能显著下降，并增加了炎症介质的释放。此外，一项经济分析表明，体外方法是目前使用的兔阴道刺激试验方法的一种经济有效的替代方法。在第二阶段，体外检测系统的用途将进一步扩大，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，以准确评估测试材料和配方的体内阴道刺激性。该方法将被转移到外部合同测试实验室，并将进行多实验室GLP验证研究。此外，还将评估该检测方法的短期和长期重复性，并将开发高通量版本的检测方法。与公共健康相关：评估化学品和配方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、杀菌剂、避孕药具、赋形剂开发和局部药物产品的行业中得到广泛应用。对阴道刺激性的评估对于将对数百万女性的化学危害降至最低很重要。所提出的基于人体重建组织的系统将为筛选具有阴道刺激性潜力的化学品/制剂提供一种灵敏和有效的检测方法。此外，该检测方法将具有成本效益，并减少使用实验室动物进行实验。

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.