Cytotoxicity and Genotoxicity of E-Cigarette Generated Aerosols Containing Diverse Flavoring Products and Nicotine in Oral Epithelial Cell Lines.
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ABSTRACT:
SUBMITTER: Tellez CS
PROVIDER: S-EPMC7846129 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:BACKGROUND:Flavoring chemicals, or flavorants, have been used in electronic cigarettes (e-cigarettes) since their inception; however, little is known about their toxicological effects. Free radicals present in e-cigarette aerosols have been shown to induce oxidative stress resulting in damage to proliferation, survival, and inflammation pathways in the cell. Aerosols generated from e-liquid solvents alone contain high levels of free radicals but few studies have looked at how these toxins are modulated by flavorants. OBJECTIVES:We investigated the effects of different flavorants on free radical production in e-cigarette aerosols. METHODS:Free radicals generated from 49 commercially available e-liquid flavors were captured and analyzed using electron paramagnetic resonance (EPR). The flavorant composition of each e-liquid was analyzed by gas chromatography mass spectroscopy (GCMS). Radical production was correlated with flavorant abundance. Ten compounds were identified and analyzed for their impact on free radical generation. RESULTS:Nearly half of the flavors modulated free radical generation. Flavorants with strong correlations included ?-damascone, ?-tetradecalactone, ?-decalactone, citral, dipentene, ethyl maltol, ethyl vanillin, ethyl vanillin PG acetal, linalool, and piperonal. Dipentene, ethyl maltol, citral, linalool, and piperonal promoted radical formation in a concentration-dependent manner. Ethyl vanillin inhibited the radical formation in a concentration dependent manner. Free radical production was closely linked with the capacity to oxidize biologically-relevant lipids. CONCLUSIONS:Our results suggest that flavoring agents play an important role in either enhancing or inhibiting the production of free radicals in flavored e-cigarette aerosols. This information is important for developing regulatory strategies aimed at reducing potential harm from e-cigarettes.
| S-EPMC5940571 | biostudies-literature
Project description:Whereas JUUL electronic cigarettes (ECs) have captured the majority of the EC market, with a large fraction of their sales going to adolescents, little is known about their cytotoxicity and potential effects on health. The purpose of this study was to determine flavor chemical and nicotine concentrations in the eight currently marketed prefilled JUUL EC cartridges ("pods") and to evaluate the cytotoxicity of the different variants (e.g., "Cool Mint" and "Crème Brulee") using in vitro assays. Nicotine and flavor chemicals were analyzed using gas chromatography-mass spectrometry in pod fluid before and after vaping and in the corresponding aerosols. 59 flavor chemicals were identified in JUUL pod fluids, and 3 were >1 mg/mL. Duplicate pods were similar in flavor chemical composition and concentration. Nicotine concentrations (average 60.9 mg/mL) were significantly higher than those of any EC products we have previously analyzed. The transfer efficiency of individual flavor chemicals that were >1 mg/mL and nicotine from the pod fluid into aerosols was generally 35-80%. All pod fluids were cytotoxic at a 1:10 dilution (10%) in the MTT and neutral red uptake assays when tested with BEAS-2B lung epithelial cells. Most aerosols were cytotoxic in these assays at concentrations between 0.2 and 1.8%. The cytotoxicity of collected aerosol materials was highly correlated with nicotine and ethyl maltol concentrations and moderately to weakly correlated with total flavor chemical concentration and menthol concentration. Our study demonstrates that (1) some JUUL flavor pods have sufficiently high concentrations of flavor chemicals that may make them attractive to youth and (2) the concentrations of nicotine and some flavor chemicals (e.g., ethyl maltol) are high enough to be cytotoxic in acute in vitro assays, emphasizing the need to determine if JUUL products will lead to adverse health effects with chronic use.
| S-EPMC6579667 | biostudies-literature
Project description:IntroductionTo examine the interaction between an added flavoring (cherry) and nicotine on the perception of electronic cigarette (e-cigarette) aerosol and how this impacts the appeal of flavored liquids for e-cigarette (e-liquids).MethodsA total of 19 subjects (13 male, 6 female) vaped six commercially available e-liquids with varying contents of nicotine (0, 6, 12 mg/mL) and cherry flavor (4.7% or 9.3% vol/vol). For each e-liquid, subjects first rated overall liking/disliking of the aerosol using the Labeled Hedonic Scale, followed by perceived intensities of sweetness, bitterness, harshness (irritation), and cherry flavor of the aerosol using the general version of Labeled Magnitude Scale.ResultsThe main findings were that (1) added nicotine increased perceived irritation and bitterness, and decreased the perceived sweetness of the e-cigarette aerosol; (2) cherry flavoring added a characteristic "cherry flavor" and an increase in the flavoring concentration from 4.7% to 9.3% tended to increase perceived intensities of sweetness, harshness, and bitterness; and (3) hedonic ratings of the e-cigarette aerosol decreased as nicotine level increased, but were not affected by flavor level.ConclusionsOur findings indicate that the appeal of the e-cigarette aerosol decreases as nicotine concentration increases. Conversely, perceived sweetness improved liking. An increase in the concentration of cherry flavoring did not appear to impact any of the measured attributes to a significant degree.ImplicationsThis work demonstrates that the perception of specific sensory attributes of e-cigarettes and their overall appeal are affected by the e-liquid constituents. Most significantly, the results suggest that nicotine decreases the sensory appeal of e-cigarettes by contributing to the perceived irritation and bitterness of the aerosol. These data have implications for the role that nicotine plays in the sensory perception and appeal of e-cigarettes aerosol and further how these sensory factors can be modulated by sweet flavoring.
| S-EPMC7171271 | biostudies-literature
Project description:BACKGROUND:As thousands of electronic cigarette (e-cigarette) refill fluids continue to be formulated and distributed, there is a growing need to understand the cytotoxicity of the flavouring chemicals and solvents used in these products to ensure they are safe. The purpose of this study was to compare the cytotoxicity of e-cigarette refill fluids/solvents and their corresponding aerosols using in vitro cultured cells. METHODS:E-cigarette refill fluids and do-it-yourself products were screened in liquid and aerosol form for cytotoxicity using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The sensitivity of human pulmonary fibroblasts, lung epithelial cells (A549) and human embryonic stem cells to liquids and aerosols was compared. Aerosols were produced using Johnson Creek's Vea cartomizer style e-cigarette. RESULTS:A hierarchy of potency was established for the aerosolised products. Our data show that (1) e-cigarette aerosols can produce cytotoxic effects in cultured cells, (2) four patterns of cytotoxicity were found when comparing refill fluids and their corresponding aerosols, (3) fluids accurately predicted aerosol cytotoxicity 74% of the time, (4) stem cells were often more sensitive to aerosols than differentiated cells and (5) 91% of the aerosols made from refill fluids containing only glycerin were cytotoxic, even when produced at a low voltage. CONCLUSIONS:Our data show that various flavours/brands of e-cigarette refill fluids and their aerosols are cytotoxic and demonstrate the need for further evaluation of e-cigarette products to better understand their potential health effects.
| S-EPMC6397802 | biostudies-other
Project description:Objectives:We compared nicotine concentrations in one brand of refill fluids that were purchased in 4 countries and labeled 0 mg of nicotine/mL. We then identified counterfeit e-cigarette products from these countries. Methods:Overall, 125 e-cigarette refill fluids were purchased in Nigeria, the United States (US), England, and China. Nicotine concentrations were measured using high performance liquid chromatography and compared to labeled concentrations. Refill fluids were examined to identify physical differences and grouped into authentic and counterfeit products. Results:Whereas nicotine was in 51.7% (15/29) of the Nigerian, 3.7% (1/27) of the Chinese and 1.6% (1/61) of the American refill fluids (range = 0.4 - 20.4 mg/mL), 8 British products did not contain nicotine. Products from China, the US, and Nigeria with trace amounts of nicotine (0.4 to 0.6 mg/mL) were authentic; however, all products from Nigeria with more than 3.7 mg/mL were counterfeit. Conclusions:We introduce 2 novel issues in the e-cigarette industry, the production of counterfeit refill fluids under a brandjacked label and inclusion of nicotine in 81.3% of the counterfeit products labeled 0 mg/mL. This study emphasizes the need for better control and monitoring of nicotine containing products and sales outlets.
| S-EPMC5937541 | biostudies-literature
Project description:The electronic cigarette (e-cigarette) has emerged as popular electronic nicotine delivery devices (ENDs). However, the general safety and validity of e-cigarettes for nicotine delivery efficacy are still not well understood. This study developed a new method for efficient measurement of nicotine levels in both the liquids (e-liquids) used in e-cigarettes and the aerosols generated from the e-cigarettes. Protonation of the pyrrolidine nitrogen of nicotine molecules by addition of excess hydrochloric acid affords an aminium salt that is readily quantified by Fourier transform ion cyclotron mass spectrometry (FT-ICR-MS). The kinetics of nicotine protonation was studied using 1H NMR spectroscopy. Quantitative analyses of nicotine in commercial e-liquids and in the corresponding derived e-cigarette aerosols were carried out using direct infusion FT-ICR-MS. The 1H NMR study of nicotine protonation revealed a first order reaction and an activation energy of 30.05 kJ mol-1. The nicotine levels measured in the commercial e-liquids were within a wide and highly variable range of -2.94% to +25.20% around the manufacturer's stated values. The results indicated considerable differences between the measured levels and the advertised levels of nicotine in the e-liquids. The nicotine quantity measured in aerosols increased linearly both with nicotine level in e-liquids (same number of puffs) and with number of puffs (same e-liquids). These data show that quality control of e-liquids and use characteristics are major variables in efficacy of nicotine delivery.
| S-EPMC5703602 | biostudies-literature
Project description:Some jurisdictions have instituted limits on electronic cigarette (ECIG) liquid nicotine concentration, in an effort to control ECIG nicotine yield, and others are considering following suit. Because ECIG nicotine yield is proportional to the product of liquid nicotine concentration (milligram per millilitre) and device power (watts) regulations that limit liquid nicotine concentration may drive users to adopt higher wattage devices to obtain a desired nicotine yield. In this study we investigated, under various hypothetical regulatory limits on ECIG liquid nicotine concentration, a scenario in which a user of a common ECIG device (SMOK TF-N2) seeks to obtain in 15 puffs the nicotine emissions equivalent to one combustible cigarette (ie, 1.8 mg). We measured total aerosol and carbonyl compound (CC) yields in 15 puffs as a function of power (15-80 W) while all else was held constant. The estimated nicotine concentration needed to achieve combustible cigarette-like nicotine yield at each power level was then computed based on the measured liquid consumption. We found that for a constant nicotine yield of 1.8 mg, reducing the liquid nicotine concentration resulted in greater amount of liquid aerosolised (p<0.01) and greater CC emissions (p<0.05). Thus, if users seek a given nicotine yield, regulatory limits on nicotine concentration may have the unintended consequence of increasing exposure to aerosol and respiratory toxicants. This outcome demonstrates that attempting to control ECIG nicotine yield by regulating one factor at a time may have unintended health effects and highlights the need to consider multiple factors and outcomes simultaneously when designing regulations.
| S-EPMC9281877 | biostudies-literature
Project description:An electronic cigarette (e-cig) generates aerosols by vaporizing the e-liquid, which mainly consists of propylene glycol (PG), vegetable glycerin (VG), and nicotine. Understanding the effects of e-liquid main compositions on e-cig aerosols is important for exposure assessment. This study investigated how the PG/VG ratio and nicotine content affect e-cig aerosol emissions and dynamics. A tank-based e-cig device with 10 different flavorless e-liquid mixtures (e.g., PG/VG ratios of 0/100, 10/90, 30/70, 50/50, and 100/0 with 0.0% or 2.4% nicotine) was used to puff aerosols into a 0.46 m3 stainless steel chamber for 0.5 h. Real-time measurements of particle number concentration (PNC), fine particulate matter (PM2.5), and particle size distributions were conducted continuously throughout the puffing and the following 2-h decay period. During the decay period, particle loss rates were determined by a first-order log-linear regression and used to calculate the emission factor. The addition of nicotine in the e-liquid significantly decreased the particle number emission factor by 33%. The PM2.5 emission factor significantly decreased with greater PG content in the e-liquid. For nicotine-free e-liquids, increasing the PG/VG ratio resulted in increased particle loss rates measured by PNC and PM2.5. This pattern was not observed with nicotine in the e-liquids. The particle loss rates, however, were significantly different with and without nicotine especially when the PG/VG ratios were greater than 30/70. Compared with nonvolatile diethyl-hexyl subacute (DEHS) aerosols, e-cig particle concentration decayed faster inside the chamber, presumably due to evaporation. These results have potential implications for assessing human exposure to e-cig aerosols.
| S-EPMC7590927 | biostudies-literature
Project description:Background: The respiratory health effects of inhalation exposure to e-cigarette flavoring chemicals are not well understood. We focused our study on the immuno-toxicological and the oxidative stress effects by these e-cigarette flavoring chemicals on two types of human monocytic cell lines, Mono Mac 6 (MM6) and U937. The potential to cause oxidative stress by these flavoring chemicals was assessed by measuring the production of reactive oxygen species (ROS). We hypothesized that the flavoring chemicals used in e-juices/e-liquids induce an inflammatory response, cellular toxicity, and ROS production. Methods: Two monocytic cell types, MM6 and U937 were exposed to commonly used e-cigarette flavoring chemicals; diacetyl, cinnamaldehyde, acetoin, pentanedione, o-vanillin, maltol and coumarin at different doses between 10 and 1,000 ?M. Cell viability and the concentrations of the secreted inflammatory cytokine interleukin 8 (IL-8) were measured in the conditioned media. Cell-free ROS produced by these commonly used flavoring chemicals were also measured using a 2',7'dichlorofluorescein diacetate probe. These DCF fluorescence data were expressed as hydrogen peroxide (H2O2) equivalents. Cytotoxicity due to the exposure to selected e-liquids was assessed by cell viability and the IL-8 inflammatory cytokine response in the conditioned media. Results: Treatment of the cells with flavoring chemicals and flavored e-liquid without nicotine caused cytotoxicity dose-dependently. The exposed monocytic cells secreted interleukin 8 (IL-8) chemokine in a dose-dependent manner compared to the unexposed cell groups depicting a biologically significant inflammatory response. The measurement of cell-free ROS by the flavoring chemicals and e-liquids showed significantly increased levels of H2O2 equivalents in a dose-dependent manner compared to the control reagents. Mixing a variety of flavors resulted in greater cytotoxicity and cell-free ROS levels compared to the treatments with individual flavors, suggesting that mixing of multiple flavors of e-liquids are more harmful to the users. Conclusions: Our data suggest that the flavorings used in e-juices can trigger an inflammatory response in monocytes, mediated by ROS production, providing insights into potential pulmonary toxicity and tissue damage in e-cigarette users.
| S-EPMC5768608 | biostudies-literature
Project description:Electronic cigarettes (ECIGs) heat and vaporize a liquid mixture to produce an inhalable aerosol that can deliver nicotine to the user. The liquid mixture is typically composed of propylene glycol (PG) and vegetable glycerin (VG), in which are dissolved trace quantities of flavorants and, usually, nicotine. Due to their different chemical and thermodynamic properties, the proportions of PG and VG in the liquid solution may affect nicotine delivery and user sensory experience. In social media and popular culture, greater PG fraction is associated with greater "throat-hit", a sensation that has been attributed in cigarette smokers to increased presence of vapor-phase nicotine. VG, on the other hand, is associated with thicker and larger exhaled "clouds". In this study, we aim to investigate how PG/VG ratio influences variables that relate to nicotine delivery and plume visibility. Aerosols from varying PG/VG liquids were generated using a digitally controlled vaping instrument and a commercially available ECIG, and analyzed for nicotine content by GC-MS. Particle mass and number distribution were determined using a six-stage cascade impactor and a fast particle spectrometer (TSI EEPS), with tightly controlled dilution and sampling biases. A Mie theory model was used to compute the aerosol scattering coefficients in the visible spectrum. Decreasing the PG/VG ratio resulted in a decrease in total particulate matter (TPM) and nicotine yield (R2 > 0.9, p<.0001). Measured particle count median diameter ranged between 44-97nm, and was significantly smaller for PG liquids. Although the particle mass concentration was lower, aerosols produced using liquids that contained VG had an order of magnitude greater light scattering coefficients. These findings indicate that PG/VG ratio is a strong determinant of both nicotine delivery and user sensory experience.
| S-EPMC7453347 | biostudies-literature