JUUL e-liquid exposure elicits cytoplasmic Ca2+ responses and leads to cytotoxicity in cultured airway epithelial cells

Toxicol Lett. 2021 Feb 1:337:46-56. doi: 10.1016/j.toxlet.2020.11.017. Epub 2020 Nov 27.

Abstract

Rationale: The popularity of new and emerging tobacco products such as E-cigarettes (E-cigs) is rapidly expanding worldwide. However, uncertainties surrounding the potential health consequences due to the use of such products exist and warrant further study.

Methods: Cultured A549 and Calu-3 airway epithelia were exposed to three out of the eight types of JUUL brand e-liquids ("Mint", "Virginia Tobacco" and "Menthol", all containing 3% nicotine at 1% and 3% (vol/vol) dilutions) and assessed for viability using a resazurin-based assay. Intracellular Ca2+ levels were measured using fluorescent indicators and pro-inflammatory cytokine levels were monitored by quantitative PCR (qPCR). Cultures were also analyzed by flow cytometry to evaluate apoptotic markers and cell viability.

Results: Exposing the airway epithelial cells to the flavored JUUL e-liquids led to significant cytotoxicity, with the "Mint" flavor being the overall most cytotoxic. The "Mint" flavored e-liquid also led to significant elevations in intracellular Ca2+ and upregulation of the pro-inflammatory cytokine IL-6 and early apoptotic marker Annexin V.

Conclusions: JUUL e-liquid challenge resulted in a loss of airway epithelial cell viability, induced pro-inflammatory responses and eventually caused apoptosis.

Keywords: Airway epithelial cells; Apoptosis; Calcium (Ca(2+)) signaling; Electronic cigarettes; Pro-inflammatory cytokines; Quantitative PCR (qPCR).

MeSH terms

  • A549 Cells
  • Apoptosis / drug effects
  • Calcium / metabolism
  • Calcium Signaling / drug effects*
  • Cell Line
  • Cell Survival / drug effects*
  • Cytokines / analysis
  • Cytokines / metabolism
  • Cytoplasm / drug effects*
  • Cytoplasm / metabolism*
  • Electronic Nicotine Delivery Systems*
  • Epithelial Cells / drug effects*
  • Flavoring Agents / toxicity
  • Humans
  • Mentha
  • Nicotine / analysis
  • Respiratory Mucosa / cytology*
  • Respiratory Mucosa / drug effects

Substances

  • Cytokines
  • Flavoring Agents
  • Nicotine
  • Calcium