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）的物理和化学性质取决于轮胎的类型和产生它们的环境条件。TWP对靶人肺细胞的功能细胞行为的任何不利影响将严重取决于其相对于其来源和环境条件的物理化学。 目的：在不同条件下（包括收集持续时间、季节、天气条件、道路条件），从不同类型的轮胎上真实的时间（whist驾驶）收集TWP。为了完全表征颗粒-形状、尺寸、表面性质、化学性质为了将单培养和共培养（以模拟原位结构）中的原代和内部转化的原代人肺细胞暴露于TWP，并与未处理的对照和毒性颗粒（ZnO）、促炎剂（例如TNF）、已知的生物活性和内化颗粒（带电聚苯乙烯颗粒）进行比较。检测一系列细胞功能、代谢和完整性的标志物作为致病性的标志物。 博士的一个重要的持续组成部分将是充分利用TWP（上述方法）。细胞模型将在顶端上皮界面暴露于TWP颗粒，使用反映低、高和慢性暴露的广泛颗粒浓度，突出显示微妙的暴露效应、暴露阈值以及任何明显效应。 颗粒的吸收和易位将使用上述技术和细胞模型进行检查，正如我们以前发表的聚苯乙烯和碳纳米管。 体外模型的使用使我们能够解决目标和3R，并使用少量的TWP（毫克）;大量是昂贵的，难以产生。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.