Coolant Flavors Exacerbate Nicotine Effects in Craniofacial Cells

Electronic Nicotine Delivery Systems (ENDS) present persistent public health challenges. Marketed for cessation, they may initiate the nicotine-naïve into addiction, and regulation has not curbed the appeal of flavors. Recent policies targeting menthol cigarettes and flavored cartridge-based ENDS are being challenged as the industry pivots to "copy-cat" coolants that users perceive similarly to menthol. Building on our prior work showing deleterious effects of vape components on craniofacial-derived cells, we examined acute in vitro exposures to coolant formulations.

We assessed cell viability, apoptosis, autophagy, and senescence in murine craniofacial derived cells with and without ENDS constituents, nicotine, propylene glycol (PG), and glycerin (G).

Consistent with previous findings that nicotine increases and nicotine+PG/G decreases cell viability, we observed that menthol and synthetic coolants WS3 and WS23 produced similar toxicological profiles by MTS assay. When combined with nicotine and PG/G, these coolants further decreased viability. Senescence occurred only for menthol when combined with PG/G and/or nicotine. Apoptosis and autophagy showed no changes. These results demonstrate that WS3 and WS23 act like menthol in craniofacial-derived cells under acute exposure conditions when assessed for cell viability.

As the tobacco and ENDS industries adopt synthetic alternatives to navigate flavor regulations, safety evaluations must explicitly include menthol like ingredients. Our data provide a biologically relevant basis for including coolant analogs in regulatory toxicology based in mesenchymal and skeletal precursor cell health. These findings support inclusion of coolant flavorants in hazard screening frameworks, especially given youth driven exposure and evolving products and device chemistries.