Pulmonary response to nanomaterial-ozone exposures

纳米二氧化钛(TiO2NPs)作为颜料广泛应用于涂料、食品、化妆品等领域。每年一次 产量估计为120万吨。虽然通过皮肤或肠道接触二氧化钛纳米颗粒被认为是 加工这些材料的工人吸入肺部是无害的，这仍然是一个令人担忧的问题。最近的结果来自 佩恩实验室于2018年1月从佐治亚理工学院迁至杜克大学，该实验室表明，二氧化钛 在没有光的情况下，NPS会产生活性氧物种(ROS)，使与蛋白质接触的蛋白质氧化 纳米二氧化钛。氧化的蛋白质在细胞中启动氧化应激反应。这些以前的实验是 用血清蛋白与HeLa和A549细胞进行比较。此R21的目标是将这些 肺部实验，与罗伯特·泰格博士合作，他是一名肺过敏教员， 杜克大学医学院的重症监护医学和肺部反应方面的专家 环境污染物。实验将从支气管肺泡灌洗液发展到呼吸道巨噬细胞 和上皮细胞。三种类型的二氧化钛纳米颗粒，先前在佩恩实验室中被表征，将用于 实验；工业级纳米二氧化钛，表面钝化的不产生ROS的纳米二氧化钛，以及等离子体- 处理后的纳米二氧化钛具有更高的ROS产量。这项研究的意义在于获得一种分子-- 纳米二氧化钛与肺细胞相互作用的水平、机制和理解，包括啮齿类动物的比较 和人体样本。本研究的创新之处在于一种新的体外探索细胞的方法。 对NPs的反应，包括毒性。这不仅仅是对纳米二氧化钛的影响，因为这种方法可以用来 了解细胞对一系列工业和环境NPs的反应。因为大多数NP 研究集中在不能代表体内实际情况的细胞培养条件上，这一实验设计 是一项重大的技术创新。这项研究的结果将是对二氧化钛机理的理解。 NPS在肺环境中，以确定其对人类健康的真正影响。虽然风险很大，但佩恩实验室 在模型系统中对这些NPs的研究，以及Tighe实验室在肺反应方面的专业知识 环境暴露，使拟议的工作在实验上容易处理。 0

Titanium dioxide nanoparticles (TiO2 NPs) are widely used as pigments in paints, food, and cosmetics. Annual production is estimated at 1.2 M metric tons. While TiO2 NP exposure through the skin or gut is considered harmless, inhalation into the lung by workers processing these materials remains a concern. Recent results from the Payne Lab, which relocated from Georgia Tech to Duke University in January 2018, has shown that TiO2 NPs, in the absence of light, produce reactive oxygen species (ROS) that oxidize proteins in contact with the TiO2 NPs. The oxidized proteins initiate an oxidative stress response in cells. These previous experiments were carried out using blood serum proteins and HeLa and A549 cells. The goal of this R21 is to translate these experiments to the lung, working in collaboration with Dr. Robert Tighe, a faculty member in Pulmonary, Allergy, and Critical Care Medicine at Duke’s School of Medicine and an expert in the pulmonary response to environmental pollutants. Experiments will progress from bronchoalveolar lavage fluid to airway macrophage and epithelial cells. Three types of TiO2 NPs, previously characterized in the Payne Lab, will be used for experiments; industrial-grade TiO2 NPs, surface passivated TiO2 NPs that do not produce ROS, and plasma- treated TiO2 NPs with increased ROS production. The significance of this research is obtaining a molecular- level, mechanistic, understanding of the interaction of TiO2 NPs with lung cells including a comparison of rodent and human samples. The innovative aspect of this research is a new in vitro method to explore cellular responses to NPs, including toxicity. This has implications beyond TiO2 NPs as this approach could be used to understand cellular responses to a broad array of industrial and environmental NPs. Since the majority of NP studies focus on cell culture conditions that do not represent actual in vivo conditions, this experimental design is a significant technical innovation. The outcome of this research will be a mechanistic understanding of TiO2 NPs in a pulmonary environment to define their true impact on human health. While risky, extensive Payne Lab studies of these NPs in model systems, coupled with Tighe Lab expertise in the pulmonary response to environmental exposures, makes the proposed work experimentally tractable. 0