High Content Screening in NHBE cells shows significantly reduced biological activity of flavoured e-liquids, when compared to cigarette smoke condensate

Toxicology in Vitro

Volumn 58, Issue , 2019, Pages 86-96

  Czekala, Lukasz ^a   Simms, Liam ^a   Stevenson, Matthew ^a   Trelles Sticken, Edgar ^b   Walker, Paul ^c   Walele, Tanvir ^a  

^a IMPERIAL COLLEGE LONDON   (United Kingdom)

^b An Imperial Brands PLC Company   (Germany)

^c NONE   (United Kingdom)

Author keywords

Cigarette; E liquids; Flavours; High Content Screening; In vitro; Normal Human Bronchial Epithelial cells

Indexed keywords

ADENOSINE TRIPHOSPHATE; CASPASE 3; CASPASE 7; CIGARETTE SMOKE CONDENSATE; E LIQUID; ENVIRONMENTAL, INDUSTRIAL AND DOMESTIC CHEMICALS; FLAVORING AGENT; GLUTATHIONE; IMMUNOGLOBULIN ENHANCER BINDING PROTEIN; NICOTINE; REACTIVE OXYGEN METABOLITE; UNCLASSIFIED DRUG; GLYCEROL; PROPYLENE GLYCOL;

APOPTOSIS; ARTICLE; CELL COUNT; CELL CYCLE ARREST; CELL DEATH; CELL MEMBRANE PERMEABILITY; CELL VIABILITY; COMPARATIVE STUDY; CONCENTRATION (PARAMETER); CONTROLLED STUDY; CYTOTOXICITY; ENERGY YIELD; ENVIRONMENTAL EXPOSURE; ENZYME ACTIVITY; HBEC CELL LINE (BRONCHIAL EPITHELIUM); HUMAN; HUMAN CELL; MITOCHONDRIAL MEMBRANE POTENTIAL; OSMOSIS; OXIDATIVE STRESS; PROTEIN CONTENT; SCREENING TEST; ADVERSE EVENT; CELL CYCLE; CELL LINE; CELL SURVIVAL; DRUG EFFECT; ELECTRONIC CIGARETTE; EPITHELIUM CELL; HIGH THROUGHPUT SCREENING; METABOLISM; SMOKE; TOBACCO;

ADENOSINE TRIPHOSPHATE; CELL CYCLE; CELL LINE; CELL SURVIVAL; ELECTRONIC NICOTINE DELIVERY SYSTEMS; EPITHELIAL CELLS; GLUTATHIONE; GLYCEROL; HIGH-THROUGHPUT SCREENING ASSAYS; HUMANS; MEMBRANE POTENTIAL, MITOCHONDRIAL; NF-KAPPA B; NICOTINE; PROPYLENE GLYCOL; REACTIVE OXYGEN SPECIES; SMOKE; TOBACCO PRODUCTS;

EID: 85063345736     PISSN: 08872333     EISSN: 18793177     Source Type: Journal    
DOI: 10.1016/j.tiv.2019.03.018     Document Type: Article

References (67)

1. Adamson, J., et al. Nicotine quantification in vitro: a consistent dosimetry marker for e-cigarette aerosol and cigarette smoke generation. Appl. In Vitro Toxicol. 3:1 (2017), 14–27.
2. Baglole, C.J., et al. Differential induction of apoptosis by cigarette smoke extract in primary human lung fibroblast strains: implications for emphysema. Am. J. Phys. Lung Cell. Mol. Phys. 291:1 (2006), L19–L29.
3. Bahl, V., et al. Comparison of electronic cigarette refill fluid cytotoxicity using embryonic and adult models. Reprod. Toxicol. 34:4 (2012), 529–537.
4. Bals, R., Hiemstra, P.S., Innate immunity in the lung: how epithelial cells fight against respiratory pathogens. Eur. Respir. J. 23:2 (2004), 327–333.
5. Barnum, K.J., O'Connell, M.J., Cell cycle regulation by checkpoints. Methods Mol. Biol. 1170 (2014), 29–40.
6. Behar, R.Z., et al. Identification of toxicants in cinnamon-flavored electronic cigarette refill fluids. Toxicol. in Vitro 28:2 (2014), 198–208.
7. Behar, R.Z., Wang, Y., Talbot, P., Comparing the cytotoxicity of electronic cigarette fluids, aerosols and solvents. Tob. Control. 27:3 (2017), 325–333.
8. Behrsing, H., et al. Characterization of a vitrocell VC1 using nicotine dosimetry: an essential component toward standardized in vitro aerosol exposure of tobacco and next generation nicotine delivery products. Appl. In Vitro Toxicol. 4:2 (2018), 159–166.
9. Berg, N., et al. Toxicology in the 21st century – working our way towards a visionary reality. Toxicol. in Vitro 25:4 (2011), 874–881.
10. Bodas, M., et al. Nicotine exposure induces bronchial epithelial cell apoptosis and senescence via ROS mediated autophagy-impairment. Free Radic. Biol. Med. 97 (2016), 441–453.
11. Boei, J., et al. Xenobiotic metabolism in differentiated human bronchial epithelial cells. Arch. Toxicol. 91:5 (2017), 2093–2105.
12. Brown, C.J., Electronic cigarettes- product characterisation and design considerations. Tob. Control. 23 (2014), ii4–ii10.
13. Brown, J., et al. Real-world effectiveness of e-cigarettes when used to aid smoking cessation: a cross-sectional population study. Addiction 109:9 (2014), 1531–1540.
14. Bullen, C., et al. Electronic cigarettes for smoking cessation: a randomised controlled trial. Lancet 382:9905 (2013), 1629–1637.
15. Dorval, T., et al. Filling the drug discovery gap: is high-content screening the missing link?. Curr. Opin. Pharmacol. 42 (2018), 40–45.
16. Elmore, S., Apoptosis: a review of programmed cell death. Toxicol. Pathol. 35:4 (2007), 495–516.
17. Farsalinos, K.E., et al. Comparison of the cytotoxic potential of cigarette smoke and electronic cigarette vapour extract on cultured myocardial cells. Int. J. Environ. Res. Public Health 10:10 (2013), 5146–5162.
18. Farsalinos, K.E., et al. Impact of flavour variability on electronic cigarette use experience: an internet survey. Int. J. Environ. Res. Public Health 10:12 (2013), 7272–7282.
19. Fields, W.R., et al. Gene expression in normal human bronchial epithelial (NHBE) cells following in vitro exposure to cigarette smoke condensate. Toxicol. Sci. 86:1 (2005), 84–91.
20. Gairola, C., Aleem, M.I., Cigarette smoke: effect of aqueous and nonaqueous fractions on mitochondrial function. Nature 241:5387 (1973), 287–288.
21. Goniewicz, M.L., et al. Levels of selected carcinogens and toxicants in vapour from electronic cigarettes. Tob. Control. 23:2 (2014), 133–139.
22. Gonzalez-Suarez, I., et al. In vitro systems toxicology assessment of nonflavored e-cigarette liquids in primary lung epithelial cells. Appl. In Vitro Toxicol. 3:1 (2017), 41–55.
23. Hartung, T., E-cigarettes and the need and opportunities for alternatives to animal testing. ALTEX 33:3 (2016), 211–224.
24. Haussmann, H.J., Fariss, M.W., Comprehensive review of epidemiological and animal studies on the potential carcinogenic effects of nicotine per se. Crit. Rev. Toxicol. 46:8 (2016), 701–734.
25. Heck, J.D., A review and assessment of menthol employed as a cigarette flavoring ingredient. Food Chem. Toxicol. 48:Suppl. 2 (2010), S1–38.
26. Hosseini, M.-J., et al. Toxicity of cigarette smoke on isolated lung, heart, and brain mitochondria: induction of oxidative stress and cytochrome c release. Toxicol. Environ. Chem. 95:9 (2013), 1624–1637.
27. Hukkanen, J., Jacob, P. 3rd, Benowitz, N.L., Metabolism and disposition kinetics of nicotine. Pharmacol. Rev. 57:1 (2005), 79–115.
28. International Organization for Standardization, ISO 3402: 1999 Tobacco and Tobacco Products - Atmosphere for Conditioning and Testing. 1999.
29. Iskandar, A.R., et al. A framework for in vitro systems toxicology assessment of e-liquids. Toxicol. Mech. Methods 26:6 (2016), 389–413.
30. Joint FAO/WHO Expert Committee on Food Additives JECFA, Propylene Glycol. 2001.
31. Jorgensen, E.D., et al. Global gene expression analysis of human bronchial epithelial cells treated with tobacco condensates. Cell Cycle 3:9 (2004), 1154–1168.
32. Kennedy, J.R., Elliott, A.M., Cigarette smoke: the effect of residue on mitochondrial structure. Science 168:3935 (1970), 1097–1098.
33. Kitagawa, S., Li, H., Effects of removal of stratum corneum, delipidization and addition of enhancers, ethanol and l-menthol, on skin permeation of benzoic acid and its 4-n-alkyl substituents in excised guinea pig dorsal skin. Chem. Pharm. Bull. (Tokyo) 47:1 (1999), 44–47.
34. Kogel, U., et al. Biological impact of cigarette smoke compared to an aerosol produced from a prototypic modified risk tobacco product on normal human bronchial epithelial cells. Toxicol. in Vitro 29:8 (2015), 2102–2115.
35. Lee, H.C., et al. Increase of mitochondria and mitochondrial DNA in response to oxidative stress in human cells. Biochem. J. 348:Pt 2 (2000), 425–432.
36. Lee, H.J., et al. Effects of nicotine on proliferation, cell cycle, and differentiation in immortalized and malignant oral keratinocytes. J. Oral Pathol. Med. 34:7 (2005), 436–443.
37. Lerner, C.A., et al. Vapors produced by electronic cigarettes and e-juices with flavorings induce toxicity, oxidative stress, and inflammatory response in lung epithelial cells and in mouse lung. PLoS One, 10(2), 2015, e0116732.
38. Liu, T., et al. NF-κB signaling in inflammation. Signal Transduction Target. Ther., 2, 2017, 17023.
39. Marescotti, D., et al. High content screening analysis to evaluate the toxicological effects of harmful and potentially harmful constituents (HPHC). J. Vis. Exp.(111), 2016.
40. McNeill, A., et al. E-cigarettes: An Evidence Update. A Report Commissioned by Public Health England. 2015, Public Health England, London.
41. Misra, M., et al. Comparative in vitro toxicity profile of electronic and tobacco cigarettes, smokeless tobacco and nicotine replacement therapy products: e-liquids, extracts and collected aerosols. Int. J. Environ. Res. Public Health 11:11 (2014), 11325–11347.
42. O'Connell, G., et al. A randomised, open-label, cross-over clinical study to evaluate the pharmacokinetic profiles of cigarettes and e-cigarettes with nicotine salt formulations in US adult smokers. Intern. Emerg. Med., 2019, 1–9.
43. Park, E.M., Park, Y.M., Gwak, Y.S., Oxidative damage in tissues of rats exposed to cigarette smoke. Free Radic. Biol. Med. 25:1 (1998), 79–86.
44. Phillips, B., et al. Toxicity of the main electronic cigarette components, propylene glycol, glycerin, and nicotine, in Sprague-Dawley rats in a 90-day OECD inhalation study complemented by molecular endpoints. Food Chem. Toxicol. 109:Pt 1 (2017), 315–332.
45. Pickett, G., et al. Effects of 10 cigarette smoke condensates on primary human airway epithelial cells by comparative gene and cytokine expression studies. Toxicol. Sci. 114:1 (2010), 79–89.
46. Rahman, I., MacNee, W., Lung glutathione and oxidative stress: implications in cigarette smoke-induced airway disease. Am. J. Phys. 277:6 Pt 1 (1999), L1067–L1088.
47. Rahman, I., et al. Glutathione homeostasis in alveolar epithelial cells in vitro and lung in vivo under oxidative stress. Am. J. Phys. 269:3 Pt 1 (1995), L285–L292.
48. Rao, P.S.S., et al. Effects of cigarette smoke condensate on oxidative stress, apoptotic cell death, and HIV replication in human monocytic cells. PLoS One, 11(5), 2016, e0155791.
49. Rovida, C., et al. Toxicity testing in the 21st century beyond environmental chemicals. ALTEX 32:3 (2015), 171–181.
50. Rowell, T.R., Sizer, S., Tarran, R., C71 you are what breathe: Airway cell biology in COPD: Flavored E-Liquids alter cell proliferation and Calcium Signaling. Am. J. Respir. Crit. Care Med., 193, 2016, 1.
51. Rowell, T.R., et al. Flavored e-cigarette liquids reduce proliferation and viability in the CALU3 airway epithelial cell line. Am. J. Phys. Lung Cell. Mol. Phys. 313:1 (2017), L52–L66.
52. Russell, C., et al. Changing patterns of first e-cigarette flavor used and current flavors used by 20,836 adult frequent e-cigarette users in the USA. Harm Reduct. J., 15(1), 2018, 33.
53. Sassano, M.F., et al. Evaluation of e-liquid toxicity using an open-source high-throughput screening assay. PLoS Biol., 16(3), 2018, e2003904.
54. Scheffler, S., et al. Evaluation of E-cigarette liquid vapor and mainstream cigarette smoke after direct exposure of primary human bronchial epithelial cells. Int. J. Environ. Res. Public Health 12:4 (2015), 3915–3925.
55. Schneider, N.G., et al. The nicotine inhaler: clinical pharmacokinetics and comparison with other nicotine treatments. Clin. Pharmacokinet. 40:9 (2001), 661–684.
56. Sheldon, L.S., Cohen Hubal, E.A., Exposure as part of a systems approach for assessing risk. Environ. Health Perspect. 117:8 (2009), 119–1194.
57. Sherwood, C.L., Boitano, S., Airway epithelial cell exposure to distinct e-cigarette liquid flavorings reveals toxicity thresholds and activation of CFTR by the chocolate flavoring 2,5-dimethypyrazine. Respir. Res., 17(1), 2016, 57.
58. Simon, H.U., Haj-Yehia, A., Levi-Schaffer, F., Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis 5:5 (2000), 415–418.
59. Tarasov, A.I., Griffiths, E.J., Rutter, G.A., Regulation of ATP production by mitochondrial Ca2+. Cell calcium 52:1 (2012), 28–35.
60. Taylor, M., et al. Assessment of novel tobacco heating product THP1.0. Part 6: a comparative in vitro study using contemporary screening approaches. Regul. Toxicol. Pharmacol. 93 (2017), 62–70.
61. Thornberry, N.A., et al. A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J. Biol. Chem. 272:29 (1997), 17907–17911.
62. van der Toorn, M., et al. Cigarette smoke irreversibly modifies glutathione in airway epithelial cells. Am. J. Phys. Lung Cell. Mol. Phys. 293:5 (2007), L1156–L1162.
63. van der Toorn, M., et al. Cigarette smoke-induced blockade of the mitochondrial respiratory chain switches lung epithelial cell apoptosis into necrosis. Am. J. Phys. Lung Cell. Mol. Phys. 292:5 (2007), L1211–L1218.
64. Walele, T., et al. Evaluation of the safety profile of an electronic vapour product used for two years by smokers in a real-life setting. Regul. Toxicol. Pharmacol. 92 (2018), 226–238.
65. Wang, Y., et al. Menthol inhibits the proliferation and motility of prostate cancer DU145 cells. Pathol. Oncol. Res. 18:4 (2012), 903–910.
66. Werley, M.S., et al. Non-clinical safety and pharmacokinetic evaluations of propylene glycol aerosol in Sprague-Dawley rats and Beagle dogs. Toxicology 287:1–3 (2011), 76–90.
67. Zhu, S.H., et al. Four hundred and sixty brands of e-cigarettes and counting: implications for product regulation. Tob. Control. 23:Suppl. 3 (2014), iii3–9.