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

Toxicol In Vitro. 2019 Aug:58:86-96. doi: 10.1016/j.tiv.2019.03.018. Epub 2019 Mar 14.


There is scientific agreement that the detrimental effects of cigarettes are produced by the formation of Harmful and Potentially Harmful Constituents from tobacco combustion and not by nicotine. For this reason numerous public health bodies and governments worldwide have indicated that e-cigarettes have a central role to play in tobacco harm reduction. In this study, high content screening (HCS) was used to compare the effects of neat e-liquids and 3R4F reference cigarette smoke condensate (CSC), which served as a positive control, in Normal Human Bronchial Epithelial (NHBE) cells. The endpoints measured covered cellular health, energy production and oxidative stress. Base liquids, with or without nicotine, and commercial, flavoured, nicotine-containing e-liquids (CFs), had little or no effect on cell viability and most HCS endpoints even at significantly higher concentrations (typically 100 times or higher) than 3R4F CSC. CSC induced a dose-dependent decrease of cell viability and triggered the response in all HCS endpoints. Effects of CFs were typically observed at or above 1%. CF Menthol was the most active flavour, with minimum effective concentrations 43 to 659 times higher than corresponding 3R4F CSC concentrations. Our results show a lower biological activity of e-liquids compared to cigarette smoke condensate in this experimental setting, across wide range of cellular endpoints.

Keywords: Cigarette; E-liquids; Flavours; High Content Screening; In vitro; Normal Human Bronchial Epithelial cells.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Cell Cycle / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Electronic Nicotine Delivery Systems*
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism
  • Glutathione / metabolism
  • Glycerol / toxicity*
  • High-Throughput Screening Assays
  • Humans
  • Membrane Potential, Mitochondrial / drug effects
  • NF-kappa B / metabolism
  • Nicotine / toxicity*
  • Propylene Glycol / toxicity*
  • Reactive Oxygen Species / metabolism
  • Smoke / adverse effects*
  • Tobacco Products / adverse effects*


  • NF-kappa B
  • Reactive Oxygen Species
  • Smoke
  • Propylene Glycol
  • Nicotine
  • Adenosine Triphosphate
  • Glutathione
  • Glycerol