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

摘要电子烟的香精、烟草、薄荷脑和降温香精已经变得广泛流行。 申请人建议调查薄荷醇/薄荷和烟草的“化学”、“毒性”和“健康影响”。 加味电子尼古丁输送系统(END)产品。这项建议解决了毒理学问题 这些香料及其成分在正常和预先存在情况下对呼吸系统的影响 通过表征化学物质，使用体外和体内模型测定毒性，以及 识别潜在的疾病生物标记物。所有可用的薄荷醇/冷藏和烟草味的末端 将对电子液体、电子汽棒和豆荚等类别进行比较。中心假设是薄荷醇/薄荷 烟草香精含有有害的调味化学物质，会引起人体细胞和分子的不良变化。 肺组织和先前存在的情况加剧了对气味暴露的毒理学反应。这个 目的是鉴定薄荷醇/凉爽和烟草香料的成分及其肺部毒性，并 确定潜在的疾病生物标志物。目标1：确定薄荷醇、类似薄荷醇(冷却)和 烟草香料，包括调味剂和雾化形成的副产品。受欢迎 烟草和薄荷/薄荷/冷却口味的末端将被雾化，它们的化学物质将被 特色化的。目的2：测定薄荷脑及其类似物的体内外毒性和对健康的影响 (冷却)，和正常情况下EpiAirway 3D组织和小鼠(C57BL/6J和BALB/C)的烟草味末端 以及先前存在的呼吸系统疾病。EpiAirway 3D急性暴露于薄荷醇/薄荷和烟草末端 将对小鼠进行(正常和患病)和慢性暴露。使用气-液-液界面(ALI) 气雾剂EpiAirway 3D组织来自正常、COPD和哮喘供体，体外毒性将风味 下定决心。通过香烟烟雾(CS)暴露(或药物)诱导小鼠COPD和哮喘 随后长期暴露于薄荷醇/薄荷和烟草口味，体内呼吸毒理学影响 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 烟草和薄荷醇/冷却蒸汽引起的疾病的标志性生物标志物。目标3：体外和体内测定 EpiAirway中薄荷脑、类薄荷脑(降温)和烟草调味剂的体内毒性和健康效应 3D组织和小鼠(C57BL/6J和BALB/C)在正常和预先存在的呼吸条件下。已确定 薄荷脑/冷却和烟草末端中存在的调味剂将用于体外治疗急性和慢性 和活体模型。将进行剂量依赖性实验，以确定诱导毒性。 利用COPD和哮喘诱导的小鼠，将确定调味剂的毒理作用。这将是 允许应聘者识别这些调味品中的关键化学物质，这些化学物质对 肺细胞和小鼠。此外，这项研究将确定这些已识别的化学物质在呼吸作用下的影响。 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.