Studies on the contributions of smoke constituents, individually and in mixtures, in a range of in vitro bioactivity assays

• Published in: Toxicology in vitro Vol. 42; pp. 222 - 246
• Main Authors: Stabbert, Regina, Dempsey, Ruth, Diekmann, Joerg, Euchenhofer, Christian, Hagemeister, Timo, Haussmann, Hans-Juergen, Knorr, Arno, Mueller, Boris P., Pospisil, Pavel, Reininghaus, Wolf, Roemer, Ewald, Tewes, Franz J., Veltel, Detlef J.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.08.2017; Elsevier Science Ltd
• Subjects: 3T3 Cells; Aldehydes; Animals; Assaying; Biocompatibility; Biological activity; Biological Assay; Cardiovascular disease; Cardiovascular diseases; Cell Survival - drug effects; Chronic obstructive pulmonary disease; Cigarette smoke; Constituents; Cytotoxicity; Fingerprinting; Glutathione - chemistry; Hydroquinone; Lung cancer; Lung diseases; Mice; Mutagenicity; Mutagenicity Tests; Nicotiana; Obstructive lung disease; Smoke; Smoke - adverse effects; Smoke - analysis; Smoke inhalation; Smoking; Tobacco; Tobacco Products; Tobacco smoke; Tobacco Smoke Pollution - analysis; Toxicity; Vapor phases
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2017.04.003

Tobacco smoke is a complex mixture with over 8700 identified constituents. Smoking causes many diseases including lung cancer, cardiovascular disease, and chronic obstructive pulmonary disease. However, the mechanisms of how cigarette smoke impacts disease initiation or progression are not well understood and individual smoke constituents causing these effects are not generally agreed upon. The studies reported here were part of a series of investigations into the contributions of selected smoke constituents to the biological activity of cigarette smoke. In vitro cytotoxicity measured by the neutral red uptake (NRU) assay and in vitro mutagenicity determined in the Ames bacterial mutagenicity assay (BMA) were selected because these assays are known to produce reproducible, quantitative results for cigarette smoke under standardized exposure conditions. In order to determine the contribution of individual cigarette smoke constituents, a fingerprinting method was developed to semi-quantify the mainstream smoke yields. For cytotoxicity, 90% of gas vapor phase (GVP) cytotoxicity of the Kentucky Reference cigarette 1R4F was explained by 3 aldehydes and 40% of the 1R4F particulate phase cytotoxicity by 10 smoke constituents, e.g., hydroquinone. In the microsuspension version of the BMA, 4 aldehydes accounted for approximately 70% of the GVP mutagenicity. Finally, the benefits of performing such studies along with the difficulties in interpretation in the context of smoking are discussed. •90% of 2R4F gas vapor phase (GVP) cytotoxicity is explained by 3 aldehydes.•40% of the 2R4F particulate phase cytotoxicity is explained by 10 constituents.•70% of the 2R4F GVP bacterial mutagenicity is explained by 4 aldehydes.