Functional Imaging of Barrier Function in 2D and 3D epithelia

该项目旨在创建下一代药物发现平台，以非侵入性方式在上皮屏障组织的微生理模型中观察屏障功能。制药行业正越来越多地从动物研究转向基于人或人源细胞的细胞测定。原因既有伦理上的，也有实际上的，因为人类细胞更准确地模仿人类细胞组织的行为和反应。这节省了药物开发管道的时间和金钱，并通过提供有关潜在人类副作用的早期数据来帮助降低铅的风险。迫切需要下一代平台技术从“微生理”系统中提取实时数据，这些系统正在通过“组织芯片”方法开发。然而，我们目前缺乏将关键生理要求（例如孔隙率、极性、流体通路和电极通路）与以易处理的成本实现可扩展制造所需的光学透明度和小尺寸组合的联合收割机能力。该项目将开发新的成像和传感方法，以补充诺丁汉开发的技术（电阻抗成像，流体动力学拉曼成像），以监测和分析Webb实验室常规使用的上皮（皮肤）屏障组织模型中的电分子屏障功能（肺：Calu-3，眼：ARPE-19，肠道：Caco-2）。将使用行业标准方法（Transwell）的细胞系统的生理参数与本文开发的新型先进材料（如纳米多孔氮化物（NPN）膜和微孔玻璃）进行比较，并在其上建立和表征2D和3D上皮细胞培养物。金标准方法将提供定量数据，证明这些新平台在高通量环境中监测生理学的潜力。我们将利用现有的链接到UoN IP和NPN生物传感器，微孔玻璃，电阻抗成像方法，生物相容性表面涂层和微生理设备的潜在商业化途径，以生成一个先进的平台，用于在暴露于药理学（候选药物，渗透增强系统）和生理学（离子置换，渗透操作）挑战期间对生理学进行灵敏和非侵入性监测。

This project aims to create next-generation drug discovery platforms to characterise barrier function in microphysiological models of epithelial barrier tissues non-invasively. The pharmaceutical industry is increasingly shifting away from animal studies towards cell assays based on human or human-derived cells. Reasons are both ethical and practical, since human cells more accurately mimic human cell tissue behaviour and responses. This saves time and money in drug development pipelines and helps to de-risk lead advancement by providing early data on potential human side-effects. Next generation platform technologies are urgently needed to extract real-time data from "microphysiological" systems, which are being developed through "tissue on chip" approaches. However, we presently lack the ability to combine key physiological requirements such as porosity, polarity, fluidic access, and electrode access with the optical transparency and small dimensions required to achieve scalable manufacture at tractable cost. This project will develop novel imaging and sensing methods to compliment techniques developed at Nottingham (electro-impedance imaging, hydrodynamic Raman imaging) to monitor and characterise electrical molecular barrier function in epithelial (skin) barrier tissue models in routine use in the Webb lab (lung: Calu-3, eye: ARPE-19, gut: Caco-2). Physiological parameters of cell systems using industry-standard methods (Transwell) will be compared with novel advanced materials such as nanoporous nitride (NPN) membranes and microporous bioglasses developed herein, on which 2D and 3D epithelial cultures will be established and characterised. Gold-standard methods will provide quantitative data demonstrating the potential of these novel platforms for the monitoring of physiology in a high-throughput context. We will leverage existing links to UoN IP and potential commercialisation pathways for NPN biosensors, microporous glasses, electroimpedance imaging methods, biocompatible surface coatings, and microphysiological devices to generate an advanced platform for the sensitive and non-invasive monitoring of physiology during exposure to pharmacological (drug candidates, permeation enhancing systems) and physiological (ion replacement, osmotic manipulation) challenges.