Flavored e-liquids increase cytoplasmic Ca²⁺ levels in airway epithelia

E-cigarettes are noncombustible, electronic nicotine-delivery devices that aerosolize an e-liquid, i.e., nicotine, in a propylene glycol-vegetable glycerin vehicle that also contains flavors. While the effects of nicotine are relatively well understood, more information regarding the potential biological effects of the other e-liquid constituents is needed. This is a serious concern, because e-liquids are available in >7,000 distinct flavors. We previously demonstrated that many e-liquids affect cell growth/viability through an unknown mechanism. Since Ca²⁺ is a ubiquitous second messenger that regulates cell growth, we characterized the effects of e-liquids on cellular Ca²⁺ homeostasis. To better understand the extent of this effect, we screened e-liquids for their ability to alter cytosolic Ca²⁺ levels and found that 42 of 100 flavored e-liquids elicited a cellular Ca²⁺ response. Banana Pudding (BP) e-liquid, a representative e-liquid from this group, caused phospholipase C activation, endoplasmic reticulum (ER) Ca²⁺ release, store-operated Ca²⁺ entry (SOCE), and protein kinase C (PKCα) phosphorylation. However, longer exposures to BP e-liquid depleted ER Ca²⁺ stores and inhibited SOCE, suggesting that this e-liquid may alter Ca²⁺ homeostasis by short- and long-term mechanisms. Since dysregulation of Ca²⁺ signaling can cause chronic inflammation, ER stress, and abnormal cell growth, flavored e-cigarette products that can elicit cell Ca²⁺ responses should be further screened for potential toxicity.