High-resolution optical imaging assessment of microbicide toxicity

评估杀微生物剂的安全性和有效性需要评估潜在的伤害 由宫颈阴道管和直肠上皮中的杀微生物剂引起。的发展。 可用于内窥镜检查、快速和定量检查的成像技术和方案 评估局部使用杀菌剂后的组织损伤可能具有重要意义 对动物模型和临床研究中杀微生物剂的开发和测试的影响。 不幸的是，目前的成像技术，如白光阴道镜或结肠镜检查不能 提供高分辨率的上皮损伤图像，不能在表面以下探测 上皮损伤和炎症可能很明显。然而，近年来，新的成像技术 已经开发了技术，现在可以执行高分辨率成像 在活体内具有显微分辨率的上皮组织。我们的总体目标是开发一种内窥镜 基于图像的方法，可用于1)评估可能导致的伤害程度 杀菌剂和2)成像研究的结果与女性生殖器感染的易感性有关 单纯疱疹病毒2型引起的小鼠宫颈、阴道部和直肠。我们将部署新兴的高分辨率 成像方式，包括共聚焦荧光显微镜和光学相干 断层扫描(OCT)，以表征发生在宫颈，阴道， 和直肠上皮未处理的性幼稚小鼠以及已知的 刺激性杀微生物剂。这些结果将与使用井的感染敏感性相关。 描述了HSV-2感染的小鼠模型。这将使我们能够评估预测价值 共聚焦荧光和OCT成像对观察到的HSV-2敏感性的影响 杀菌剂诱导的上皮细胞改变，注意结果的重复性/一致性。 在这个项目的第一阶段，我们将展示高分辨率光学成像的能力，以 定量评估宫颈、阴道和直肠组织对已知杀菌剂和 测试预测杀菌剂诱导的敏感性变化的生物终点的能力 在宫颈阴道处。在第二阶段，拟议的基于图像的直肠反应评估将 以建立基于图像的标志物与直肠易感性之间的相关性 在使用杀微生物剂之后。此外，仪器和成像参数将 优化成像方案，使其适用于宫颈、阴道和直肠的成像 大型动物模型和人类的上皮反应。我们还计划使用开发的 评估新型杀微生物剂在小动物模型中表现的成像方案作为新的 产品是开发出来的。

Evaluation of safety and efficacy of microbicides requires an assessment of potential injury caused by micobicides in the epithelium of cervicovaginal tract and rectum. The development of imaging technology and protocols that can be used for endoscopic, rapid, and quantitative assessment of tissue injury following topical application of microbicides could have significant impact on the development and testing of microbicides in animal models and clinical studies. Unfortunately, current imaging technology such as white light colposcopy or colonoscopy cannot provide high resolution images of epithelial injury and cannot probe below the surface where epithelial injury and inflammation may be evident. However, in recent years, new imaging technology has been developed where it is now possible to perform high resolution imaging of epithelial tissue with microscopic resolution in vivo. Our overall goal is to develop an endoscopic imagedbased approach that can be used to 1) assess the degree of injury that may be induced by microbicides and 2) correlate the results of imaging studies to susceptibility to genital infection in mice cervicovaginal tract and rectum caused by HSV-2. We will deploy emerging high resolution imaging modalities including confocal fluorescence microscopy and optical coherence tomography (OCT) to characterize the changes that occur in the architecture of cervical, vaginal, and rectal epithelium of untreated sexually na¿ve mice as well as mice treated with known irritative microbicides. These results will be correlated to susceptibility to infection using a well characterized mouse model of HSV-2 infection. This will allow us to assess the predictive value of confocal fluorescence and OCT imaging for observed HSV-2 susceptibility based on microbicide-induced epithelial changes with attention to reproducibility/consistency of findings. In phase I of this project we will demonstrate the capabilities of high resolution optical imaging to quantitatively assess the response of cervicovaginal and rectal tissue to known microbicides and test the ability to predict the biological end point of microbicide-induced changes in susceptibility in cervicovaginal tract. In phase II, the proposed imagedbased assessment of rectal response will be extended to establish correlation between image-based markers and rectal susceptibility following application of microbicides. Furthermore, instrumentation and imaging parameters will be optimized to make the imaging protocol suitable for imaging of cervicovaginal and rectal epithelial response in large animal models and humans. We also plan to use the developed imaging protocol to assess the performance of novel microbicides in small animal models as new products are developed.