Pulmonary toxicological evaluation and chemical interactions of menthol, mint, and tobacco flavored e-cigarette products

电子烟香料、烟草香料、薄荷醇香料和清凉香料已经变得广泛流行。 申请人提议研究薄荷醇/薄荷和烟草的“化学”、“毒性”和“健康影响” 电子尼古丁输送系统（ENDS）本提案涉及毒理学 这些风味及其成分对正常和预先存在的呼吸系统的影响 通过表征化学品，使用体外和体内模型确定毒性，以及 识别疾病的潜在生物标志物。所有可用的薄荷/冷却和烟草味ENDS 将比较类别，即，电子烟油，电子烟棒，还有豆荚中心假设是薄荷醇/薄荷 烟草香料含有有害的香料化学物质， 肺组织和预先存在的状况加剧了暴露于香料时的毒理学反应。的 目的是鉴定薄荷醇/清凉和烟草香料的成分及其肺毒性， 确定疾病的潜在生物标志物。目的1：鉴别薄荷醇、薄荷醇样（清凉）和薄荷醇的化学性质。 烟草香料，包括香料化学品和在雾化时形成的次级产物。受欢迎 烟草和薄荷醇/薄荷/清凉香料的末端将被雾化，它们的化学物质将被 表征了目的2：测定薄荷醇、类薄荷醇的体内外毒性和健康效应 在正常条件下，EpiAirway 3D组织和小鼠（C57 BL/6 J和BALB/C）中的烟草味ENDS 和已有的呼吸系统疾病薄荷醇/薄荷和烟草急性暴露于EpiAirway 3D （正常和患病）和慢性暴露于小鼠。使用气液界面（ALI） 气溶胶EpiAirway 3D组织来自正常、COPD和哮喘供体，将研究香料的体外毒性。 测定通过香烟烟雾（CS）暴露（或药物）诱导小鼠COPD和哮喘病症， 随后长期暴露于薄荷醇/薄荷和烟草香料， 将确定正常和疾病状态。提出的体外和体内模型将为我们提供相关的 由于烟草和薄荷醇/冷却蒸汽引起的疾病的标志性生物标志物。目的3：体外和体内测定 在EpiAirway中薄荷醇、薄荷醇样（清凉）和烟草香料化学品的体内毒性和健康效应 正常和预先存在的呼吸条件下的3D组织和小鼠（C57 BL/6 J和BALB/C）。识别 薄荷醇/冷却和烟草ENDS中存在的调味化学品将用于急性和慢性体外试验， 和体内模型。将进行剂量依赖性实验以确定诱导毒性。 使用COPD和哮喘诱导的小鼠，将确定调味化学品的毒理学效应。这将 允许候选人识别这些调味ENDS中导致不良影响的关键化学物质， 肺细胞和小鼠。此外，这项研究将确定这些已确定的化学品在呼吸系统中的影响。 疾病状况和疾病恶化的生物标志物。

SUMMARY Electronic cigarette flavors, tobacco, menthol, and cooling flavors have become widely popular. The applicant proposes to investigate “Chemistry”, “Toxicity”, and “Health effects” of menthol/mint and tobacco flavored electronic nicotine delivery systems (ENDS) products. This proposal addresses the toxicological implications of these flavors and their constituents on the respiratory system under normal and pre-existing conditions by characterizing chemicals, determining the toxicity using in vitro and in vivo models, and identifying potential biomarkers of disease. Menthol/cooling and tobacco flavored ENDS from all available categories will be compared, i.e., e-liquids, vape bars, and pods. The central hypothesis is that menthol/mint and tobacco flavors contain harmful flavoring chemicals inducing adverse cellular and molecular changes in the lung tissue and pre-existing conditions exacerbate the toxicological response upon flavor exposures. The Goal is to identify constituents of menthol/cooling and tobacco flavors and their pulmonary toxicity and to determine potential biomarkers of disease. Aim 1: Identify the chemistry of menthol, menthol-like (cooling), and tobacco flavors, including flavoring chemicals and secondary products formed upon aerosolization. Popular ENDS of tobacco and menthol/mint/cooling flavors will be aerosolized and their chemicals will be characterized. Aim 2: Determine in vitro and in vivo toxicity and health effects of menthol, menthol-like (cooling), and tobacco flavored ENDS in EpiAirway 3D tissues and mice (C57BL/6J and BALB/C) under normal and pre-existing respiratory conditions. Acute exposure of menthol/mint and tobacco ENDS to EpiAirway3D (normal and diseased) and chronic exposure to mice will be conducted. Using an air-liquid-interface (ALI) aerosol EpiAirway 3D tissues from normal, COPD, and asthmatic donors, in vitro toxicity of flavors will be determined. By inducing COPD and asthma conditions in mice by cigarette smoke (CS) exposure (or drug) and subsequent chronic exposure to menthol/mint and tobacco flavors, in vivo respiratory toxicological effects in normal and disease states will be determined. Proposed in vitro and in vivo models will provide us relevant signature biomarkers of disease due to tobacco and menthol/cooling vapors. Aim 3: Determine in vitro and in vivo toxicity and health effects of menthol, menthol-like (cooling), and tobacco flavoring chemicals in EpiAirway 3D tissues and mice (C57BL/6J and BALB/C) under normal and pre-existing respiratory conditions. Identified flavoring chemicals present in menthol/cooling and tobacco ENDS will be used for acute and chronic in vitro and in vivo models, respectively. Dose-dependent experiments will be conducted to determine induced toxicity. Using COPD and asthma induced mice, toxicological effects of flavoring chemicals will be determined. This will allow the candidate to identify key chemicals in these flavored ENDS responsible for causing adverse effects in lung cells and mice. Further, this study will determine the effects of these identified chemicals under respiratory disease conditions and biomarkers of disease exacerbation.