Determination of the effect of inhaled tyre wear particles on the function and integrity of the human gas-blood barrier

轮胎磨损颗粒（TWP）表现出物理和化学性质，这取决于轮胎的类型和产生它们的环境条件。TWP对目标人肺细胞功能细胞行为的任何不利影响将主要取决于其相对于来源和环境条件的物理化学性质。目标：在不同条件下（包括收集时间、季节、天气条件、道路条件），实时收集不同类型轮胎的TWP（在驾驶时）。将单培养和共培养（模拟原位结构）的原代和内部转化的原代人肺细胞暴露于TWP中，并与未处理的对照和有毒颗粒（氧化锌），促炎剂（如TNF），已知的生物反应性颗粒和内化颗粒（带电聚苯乙烯颗粒）进行比较。测量一系列细胞功能、代谢和完整性的标志物，作为致病性的标志物。测量颗粒在上皮-内皮气血屏障上的摄取和易位。博士学位的一个重要组成部分将是全面描述twp（上述方法）。细胞模型将暴露在顶端上皮界面的TWP颗粒中，使用反映低、高和慢性暴露的大范围颗粒浓度，突出显示细微的暴露效应、暴露阈值以及任何明显的影响。颗粒摄取和转运将使用上述技术和细胞模型进行检查，正如我们之前发表的聚苯乙烯和碳纳米管。使用体外模型使我们能够解决目标和3r，并使用少量twp（毫克）；更大的数额是昂贵的，而且很难产生。这些数据将为未来的体内研究提供信息。

Tyre wear particles (TWP) exhibit physical and chemical properties that depend on the type of tyre and the environmental conditions in which they are generated. Any adverse effects of TWP on the functional-cellular behaviour of target human lung cells will critically depend on their physicochemistry relative to their source and environmental conditions. Objectives: To collect TWP in real time (whist driving) from different types of tyres, under varying conditions (including duration of collection, season, weather conditions, road conditions). To fully characterise the particles - shape, size, surface properties, chemistry To expose primary and in-house transformed primary human lung cells in monoculture and co-culture (to mimic in situ structure) to the TWP, and compare with untreated control and toxic particles (ZnO), pro-inflammatory agents (eg TNF), known bioreactive and internalised particles (charged polystyrene particles). To measure a series of markers of cell function, metabolism and integrity as markers of pathogenicity To measure particle uptake and translocation across epithelial-endothelial gas-blood barrier. An important ongoing component of the PhD will be to fully characterise TWPs (methods above). Cell models will be exposed to TWP particles at the apical epithelial interface, using a wide range of particle concentrations reflecting low, high and chronic exposures, highlighting subtle exposure effects, thresholds of exposure and also any overt effects. Particle uptake and translocation will be examined using the techniques and cell models described above as we have previously published for polystyrene and carbon nanotubes. Use of in vitro models allows us to address the objectives and the 3Rs, and uses small amounts of TWPs (milligrams); larger amounts are costly and difficult to generate. These data will inform future in vivo studies.