Cao Xuefei, Muskhelishvili Levan, Latendresse John, Richter Patricia, Heflich Robert H
U.S. Food and Drug Administration/National Center for Toxicological Research, Jefferson, Arkansas 72079.
Toxicologic Pathology Associates, Jefferson, Arkansas 72079.
Toxicol Sci. 2017 Mar 1;156(1):14-24. doi: 10.1093/toxsci/kfw239.
Exposure to cigarette smoke causes a multitude of pathological changes leading to tissue damage and disease. Quantifying such changes in highly differentiated in vitro human tissue models may assist in evaluating the toxicity of tobacco products. In this methods development study, well-differentiated human air-liquid-interface (ALI) in vitro airway tissue models were used to assess toxicological endpoints relevant to tobacco smoke exposure. Whole mainstream smoke solutions (WSSs) were prepared from 2 commercial cigarettes (R60 and S60) that differ in smoke constituents when machine-smoked under International Organization for Standardization conditions. The airway tissue models were exposed apically to WSSs 4-h per day for 1-5 days. Cytotoxicity, tissue barrier integrity, oxidative stress, mucin secretion, and matrix metalloproteinase (MMP) excretion were measured. The treatments were not cytotoxic and had marginal effects on tissue barrier properties; however, other endpoints responded in time- and dose-dependent manners, with the R60 resulting in higher levels of response than the S60 for many endpoints. Based on the lowest effect dose, differences in response to the WSSs were observed for mucin induction and MMP secretion. Mitigation of mucin induction by cotreatment of cultures with N-acetylcysteine suggests that oxidative stress contributes to mucus hypersecretion. Overall, these preliminary results suggest that quantifying disease-relevant endpoints using ALI airway models is a potential tool for tobacco product toxicity evaluation. Additional research using tobacco samples generated under smoking machine conditions that more closely approximate human smoking patterns will inform further methods development.
接触香烟烟雾会导致多种病理变化，进而造成组织损伤和疾病。在高度分化的体外人体组织模型中量化此类变化，可能有助于评估烟草产品的毒性。在这项方法开发研究中，使用了分化良好的人体气液界面（ALI）体外气道组织模型来评估与烟草烟雾暴露相关的毒理学终点。根据国际标准化组织的条件，对两种商业香烟（R60和S60）进行机器抽吸，制备出全主流烟雾溶液（WSS），这两种香烟的烟雾成分有所不同。气道组织模型每天经顶部分别暴露于WSS中4小时，持续1至5天。检测细胞毒性、组织屏障完整性、氧化应激、粘蛋白分泌和基质金属蛋白酶（MMP）排泄情况。这些处理没有细胞毒性，对组织屏障特性的影响微乎其微；然而，其他终点呈现出时间和剂量依赖性反应，在许多终点上，R60引起的反应水平高于S60。基于最低效应剂量，观察到在粘蛋白诱导和MMP分泌方面对WSS的反应存在差异。用N - 乙酰半胱氨酸共同处理培养物可减轻粘蛋白诱导，这表明氧化应激会导致粘液分泌过多。总体而言，这些初步结果表明，使用ALI气道模型量化与疾病相关的终点是评估烟草产品毒性的一种潜在工具。使用在吸烟机条件下产生的、更接近人类吸烟模式的烟草样品进行的进一步研究，将为后续方法开发提供信息。
