A novel clinical method to measure skin staining reveals activation of skin damage pathways by cigarette smoke.

BACKGROUND: Long-term use of cigarettes can result in localised staining and aging of smokers' skin. The use of tobacco heating products (THPs) and electronic cigarettes (ECs) has grown on a global scale; however, the long-term effect of these products' aerosols on consumers' skin is unknown. This pilot clinical study aimed to determine whether THP or EC aerosol exposure results in skin staining or activation of biomarkers associated with oxidative stress. MATERIALS AND METHODS: Eight areas were identified on the backs of 10 subjects. Two areas were used for air control, and two areas exposed to 32-puffs of cigarette smoke (CS), THP or EC aerosols, which were delivered to the skin using a 3-cm diameter exposure chamber and smoke engine. Skin colour was measured using a Chromameter. Squalene (SQ), SQ monohydroperoxide (SQOOH) and malondialdehyde (MDA) levels were measured in sebum samples by mass spectrometry and catalase colorimetry. RESULTS: CS exposure significantly increased skin staining, SQOOH and MDA levels and SQOOH/SQ ratio. THP and EC values were significantly lower than CS; EC values being comparable to air control. THP values were comparable to EC and air control at all endpoints, apart from skin staining. SQ and catalase levels did not change with exposure. CONCLUSIONS: CS stained skin and activated pathways known to be associated with skin damage. THPs and ECs produced significantly lower values, suggesting they could offer hygiene and cosmetic benefits for consumers who switch exclusively from smoking cigarettes. Further studies are required to assess longer-term effects of ECs and THPs on skin function.

Enamel staining with e-cigarettes, tobacco heating products and modern oral nicotine products compared with cigarettes and snus: An in vitro study.

PURPOSE: To evaluate the effect of cigarette smoke, smokeless tobacco (e.g. snus), tobacco heating products (THP), electronic cigarettes (EC), and modern oral nicotine products on tooth staining. METHODS: In this in vitro study, staining was assessed for 86 days following exposure of bovine enamel samples to a scientific reference cigarette (1R6F), a THP (glo), an EC (ePen 3), a reference snus product (CRP1.1), and a modern oral product (LYFT). Red wine and coffee were used as positive controls and DMSO and complete artificial saliva as negative controls. Whether brushing could reduce staining levels was also assessed. Changes in staining levels were assessed using the Commission Internationale de L'éclairage L*a*b* method. RESULTS: Enamel staining increased with incubation time, and cigarette smoke, snus, coffee and wine induced statistically higher staining levels. THP, EC and modern oral exposure induced minimal staining levels that were also comparable to negative control samples. At day 86, ΔE mean and SD values were 28.50 ± 3.14, 19.76 ± 1.26, 17.35 ± 3.44, 16.22 ± 2.07, 18.30 ± 3.82, 4.10 ± 1.99, 11.30 ± 2.60, 49.56 ± 2.44 for cigarette, glo, EC with blended tobacco, EC with rich tobacco, reference snus product, modern oral product, coffee or wine. The control ΔE mean and SD values at day 86 were 18.68 ± 3.89 for DMSO and 2.17± 0.78 for complete artificial saliva. The ΔE values for all DMSO extracted samples and control increased from day 1 to 86, which suggests that the DMSO used to extract the samples contributes to the enamel sample staining levels. Staining levels were reduced by brushing. CLINICAL SIGNIFICANCE: Cigarette smoke, red wine, snus and coffee stained enamel. Exposure to THP, EC or modern oral product extracts for 86 days resulted in minimal enamel staining. Further studies are required to assess the long-term impact on staining and the oral cavity following consumer exclusive use of EC, THP or modern oral products.

Assessment of priority tobacco additives per the requirements of the EU Tobacco Products Directive (2014/40/EU): Part 3, Smoking behavior and plasma nicotine pharmacokinetics.

This publication is part of a series of 3 publications and describes the clinical assessment performed to fulfill the regulatory requirement per Art. 6 (2) of the EU Tobacco Products Directive 2014/40/EU under which Member States require manufacturers and importers of cigarettes and Roll Your Own tobacco containing an additive that is included in the priority list established by Commission Implementing Decision (EU) 2016/787 to carry out comprehensive studies (European Union, 2016). In our clinical study, two distinct end points were investigated, namely measuring plasma nicotine pharmacokinetics as a measure of nicotine uptake, and analyses of changes in smoker puffing behavior as a measure of cigarette smoke inhalation. This clinical study indicated that the inclusion of none of the priority additives either as single additive or as part of a chemical mixture, facilitated nicotine uptake. Furthermore, the data did not suggest that differences in the inhalation pattern of cigarette smoke of any of the Priority Additives tested occurred when compared to the additive-free reference cigarette. Finally, it is concluded that neither the scientific literature nor our study gave circumstantial indications of increased addictiveness for cigarettes containing these priority additives.

Fractionation of cadmium in tobacco and cigarette smoke condensate using XANES and sequential leaching with ICP-MS/MS.

Fractionation data for cadmium in tobacco products, as obtained by sequential leaching of cadmium species with ICP-MS/MS analysis, and separately by X-ray absorption near edge structure (XANES) are presented here for the first time. The total amount of cadmium found in 3R4F cigarette cut tobacco was 1526 ± 42 µg kg-1, of which 5% was found in the smoke under ISO smoking conditions. XANES analysis showed that Cd in tobacco, cigarette smoke and ash was present in the + 2 oxidation state. Examination of the gas-particle partitioning of smoke cadmium suggests that Cd in mainstream smoke is best viewed as semi-volatile, existing in both particulate and gas phases. Sequential extraction of trapped tobacco smoke was carried out to get a deeper insight into the chemistry of cigarette smoke cadmium compounds. Consecutive extractions with ultrapure water, dilute (1%) nitric acid and 10% nitric acid led to extraction of a total amount of Cd which agreed with that obtained after microwave digestion of the whole sample, suggesting that cadmium was quantitatively leachable into aqueous/acidic solutions. Most Cd (~ 90% of the total Cd in the smoke condensate) was extracted into dilute nitric acid (likely as CdO, Cd(OH)2 and CdCO3) with a minor percentage (3%) extracted into water (likely as CdCl2) and in 10% nitric acid (likely as CdS). Extraction of trapped mainstream smoke with pentane, followed by ICP-MS/MS analysis, to examine the possible presence of organocadmium in 3R4F tobacco smoke, did not show the presence of organocadmium compounds above the method LOQ (2 µg kg-1), possibly due to their reactivity under the experimental conditions. The high selectivity with sufficient sensitivity achieved by ICP-MS/MS was invaluable to quantify Cd (at low µg kg-1levels) simultaneously with sulphur and chlorine in the tobacco smoke fractions of complex matrix. The cadmium chemistry in the smoke, identified in this study, is consistent with both relatively high lung absorption and DNA binding; both potentially important factors for disease progression in smokers. Graphical Abstract This paper provides quantitative fractionation data for cadmium in tobacco and smoke by using sequential leaching with ICPMS and XANES.

Assessment of enamel discoloration in vitro following exposure to cigarette smoke and emissions from novel vapor and tobacco heating products.

PURPOSE: To evaluate in vitro enamel sample discoloration following exposure to a scientific reference cigarette (3R4F) or emissions from next generation tobacco and nicotine products (NGPs) such as electronic cigarettes (EC) and tobacco heating products (THP). METHODS: Bovine enamel blocks (6.5 × 6.5 mm) were prepared and pre-incubated with human or artificial saliva, to form a pellicle layer before exposure to either particulate matter (PM) or whole aerosols. PM was prepared by capturing 3R4F cigarette smoke (CS), a commercial THP (THP1.0) or a novel vapor product (NVP)/next generation e-cigarette aerosols on Cambridge filter pads followed by elution with dimethyl sulfoxide (DMSO). Ten enamel samples were exposed to each PM for 14 days. For aerosol exposure, 12 enamel samples were exposed (200 puffs per day, for 5 consecutive days) to 3R4F CS or THP1.0 and NVP aerosols. Control samples were incubated with DMSO (PM study) or phosphate buffered saline (PBS, aerosol study). Individual enamel sample color readings (L*, a*, b*) were measured at baseline and on each exposure day. Mean ΔL*, Δa*, Δb* and ΔE values were calculated for each product or control. A one-way ANOVA was used to assess the differences between the products and controls. The Tukey procedure for pairwise comparisons was also used. RESULTS: At all timepoints, 3R4F PM and CS induced enamel discoloration that was statistically significant (< 0.0001) when compared to THP1.0 or NVP. After 14-day PM exposure, mean ΔE values were 29.4± 3.6, 10.5 ± 2.3, 10.7 ± 2.6 and 12.6 ± 2.0 for 3R4F, THP1.0, NVP and DMSO control respectively. After 5-day CS or aerosol exposure, mean ΔE values were 26.2 ± 3.2, 3.6 ± 1.9, 3.4 ± 1.3, 5.3 ± 0.8 for 3R4F CS, THP1.0, NVP or PBS control, respectively. Both exposure methods demonstrated that THP1.0 and NVP induced minimal staining, mean ΔL* , Δa* , Δb* and ΔE values were comparable to DMSO or PBS controls. CLINICAL SIGNIFICANCE: For the first time, diverse NGPs across the risk continuum were assessed in vitro for their impact on enamel staining. CS exposure significantly increased the level of bovine enamel sample discoloration, whereas THP1.0 or NVP exposure resulted in values comparable to the controls.

Pilot study to determine differences in breath odour between cigarette and e-cigarette consumers.

Cigarette smoke is known to influence breath odour, but the effect of e-cigarettes is unknown. In this pilot study, we aimed to determine differences in breath odour between cigarette smokers (CSs) and e-cigarette consumers (ECs) in 33 healthy subjects: 11 CSs, 11 ECs and 11 non-smokers (NSs). Breath was sampled at baseline and following product use (CSs and ECs) or a waiting period (NSs) by eight trained odour judges using a six-point smoke intensity scale and a nine-point hedonic scale. We observed a statistically significant difference between CSs and ECs. Smoke intensity values were significantly higher in CSs than ECs and NSs, which were comparable both at baseline and after product use. In addition, hedonic values for CSs were significantly lower than both NSs and ECs after product use. These acute results indicate that cigarette and e-cigarette use results in significantly different breath. ECs breath has a reduced smoke odour and more pleasant aroma than CSs, and is comparable to NSs. This suggests there may be cosmetic benefits for CSs who quit smoking or switch to exclusive use of ECs. Further studies are required to understand the long-term effects of e-cigarette use on breath odour.

Development of a novel method to measure material surface staining by cigarette, e-cigarette or tobacco heating product aerosols.

Tobacco smoke (CS) may visually stain indoor surfaces including ceilings, walls and soft furnishings over time. Potentially reduced risk products (PRRPs) such as e-cigarettes (EC) and tobacco heating products (THP) produce chemically less complex aerosols with significantly reduced levels of toxicants, particles and odour. However, the potential effects of EC and THP aerosols on the staining of indoor surfaces are currently unknown. In this study, an exposure chamber was developed as a model system to enable the accelerated staining of wallpaper and cotton samples by a scientific reference cigarette (3R4F), three THP (glo™, glo™ pro, glo™ sens) and an e-cigarette (iSwitch Maxx). Exposure to 3R4F reference cigarettes caused the greatest level of staining, which was significantly higher than glo™, glo™ pro, glo™ sens or iSwitch Maxx aerosols, all of which showed relatively little colour change. Exposure to 200-1000 puffs of 3R4F cigarette smoke resulted in a visible dose response effect to wallpaper and cotton samples which was not observed following exposure to glo™, glo™ pro, glo™ sens or iSwitch Maxx aerosols. Aging of the samples for 4 weeks post-exposure resulted in changes to the staining levels, however PRRP staining levels were minimal and significantly lower than 3R4F exposed samples. For the first time, diverse PRRPs across the tobacco and nicotine products risk continuum have been assessed in vitro for their impact on surface staining. CS exposure significantly increased the level of wallpaper and cotton staining, whereas exposure to glo™, glo™ pro, glo™ sens or iSwitch Maxx aerosols resulted in significantly reduced levels of staining, staining levels were also comparable to untreated control samples.