Novel instrument to generate representative e-cigarette vapors for physicochemical particle characterization and in-vitro toxicity

The use of “electronic cigarettes” (e-cigs) has exponentially increased during the last decade. This drastic growth has been observed in smokers and never smokers, women and teenagers, with indicators for adverse effects related to nicotine, but also to e-liquid and vapor components. The aerosol characteristics depend on the e-cig features. Therefore, the introduction of many different e-cig types on the market leads to concerns regarding their potential health effects. Our study aims to develop an instrument to generate representative vapors from e-cigarette devices, characterize the physicochemical properties of the aerosols and elucidate the adverse effects of inhaled vapors on normal human airway epithelia. E-cig vapors were produced using a specially developed aerosol generation instrument mimicking user puffing behavior. Aerosol particle size distribution and concentration were characterized with a Scanning Mobility Particle Sizer. Air-liquid interface cultures of differentiated human bronchial epithelial cells were exposed to 1 puff topography, i.e. to 21 puffs of 3 s with 22-second puff intervals using the Nano-Aerosol Chamber for In-Vitro Toxicity, which simulates particle deposition in the respiratory tract. Cell morphology was evaluated by light microcopy and induction of cell death by release of lactate dehydrogenase at 4 and 24 h after aerosol exposure. Particle size in e-cig generated aerosols ranged from 20 to 430 nm and concentrations exceeded 1 × 106 particles/cm3. Deposited particle doses were 0.9–3 μg/cm2, depending on e-device type and number of puffs. A single exposure to e-cig vapors caused changes in epithelial cell shape but no massive cell death. We successfully developed an aerosol generation system to produce and deposit realistic vapors from e-cigs. The results obtained indicate that a single exposure to e-cig aerosols affects epithelial morphology and slightly increases cell death. Our interdisciplinary approach combining aerosol technology and biology allows identifying adverse effects of e-cigs to the respiratory tract in vitro.