Tobacco Heating System Research Articles

Impacts of exhaled aerosol from the usage of the tobacco heating system to indoor air quality: A chamber study

Aerosol particle, carbonyl, and nicotine concentrations were analysed as pollutants affecting indoor air quality during the usage of electrically-heated tobacco product - the Tobacco Heating System (THS). Quantitative experimental variables included THS use intensity as number of parallel users (1, 3, or 5), distance to the bystander (0.5, 1, or 2 m), as well as environmental conditions in a chamber: ventilation intensity as air changes per hour (0.2, 0.5, or 1 h−1), and relative humidity (RH, 30, 50 or 70%). The real-time particle number (PNC), CO and CO2 concentration, as well as off-line acetaldehyde, formaldehyde, nicotine, and 3-ethenylpyridine concentration was measured during and after the active usage. Use of THS resulted in a statistically significant increase of several analytes including nicotine, acetaldehyde, PM2.5, and PNC as compared to the background. The obtained levels were significantly lower (approximately 16, 8, 8 and 28 times for nicotine, acetaldehyde, PNC and PM2.5, respectively) compared to the levels resulting from conventional cigarette (CC) smoking under identical conditions. The maximum 30 min concentration of PNC (4.8 × 105 #/cm3), as well as maximum concentration of PNC (9.3 × 106 #/cm3) suggest that the intensive use of THS in a confined space with limited ventilation might cause substantially elevated aerosol concentrations, although these particles appeared as highly volatile ones and evaporated within seconds. Generally, the usage intensity (number of simultaneous users) prevailed as the most important factor positively affecting pollutant variations; another important factor was the distance to bystander.

A six-month systems toxicology inhalation/cessation study in ApoE−/− mice to investigate cardiovascular and respiratory exposure effects of modified risk tobacco products, CHTP 1.2 and THS 2.2, compared with conventional cigarettes

Smoking is one of the major modifiable risk factors in the development and progression of chronic obstructive pulmonary disease (COPD) and cardiovascular disease (CVD). Modified-risk tobacco products (MRTP) are being developed to provide substitute products for smokers who are unable or unwilling to quit, to lessen the smoking-related health risks. In this study, the ApoE-/- mouse model was used to investigate the impact of cigarette smoke (CS) from the reference cigarette 3R4F, or aerosol from two potential MRTPs based on the heat-not-burn principle, carbon heated tobacco product 1.2 (CHTP1.2) and tobacco heating system 2.2 (THS 2.2), on the cardiorespiratory system over a 6-month period. In addition, cessation or switching to CHTP1.2 after 3 months of CS exposure was assessed. A systems toxicology approach combining physiology, histology and molecular measurements was used to evaluate the impact of MRTP aerosols in comparison to CS. CHTP1.2 and THS2.2 aerosols, compared with CS, demonstrated lower impact on the cardiorespiratory system, including low to absent lung inflammation and emphysematous changes, and reduced atherosclerotic plaque formation. Molecular analyses confirmed the lower engagement of pathological mechanisms by MRTP aerosols than CS. Both cessation and switching to CHTP1.2 reduced the observed CS effects to almost sham exposure levels.

Reduction in Exposure to Selected Harmful and Potentially Harmful Constituents Approaching Those Observed Upon Smoking Abstinence in Smokers Switching to the Menthol Tobacco Heating System 2.2 for 3 Months (Part 1).

IntroductionThe Tobacco Heating System (THS) is a “heat-not-burn” tobacco product designed to generate significantly lower levels of harmful and potentially harmful constituents (HPHCs) and present lower risk of harm than cigarettes. This study assessed the exposure reduction to selected HPHCs in smokers switching to menthol Tobacco Heating System (mTHS) 2.2 compared with smokers continuing smoking menthol cigarettes (mCCs) and smoking abstinence (SA) for 5 days in a confined setting, followed by an 86-day ambulatory period.MethodsA total of 160 healthy adult US smokers participated in this randomized, three-arm parallel group, controlled clinical study. Biomarkers of exposure to 16 HPHCs were measured in blood and 24-hour urine. Safety was monitored throughout the study. Information was also gathered on product evaluation, product use, subjective effects, and clinical risk markers (co-publication Part 2).ResultsNicotine uptake was comparable in both exposure groups (mTHS:mCC ratio of 96% on day 90). On day 5, biomarker of exposure levels to other HPHCs were reduced by 51%–96% in the mTHS group compared with the mCC group, and these reductions were sustained for most biomarkers of exposure over ambulatory period. After 90 days of use, the level of satisfaction with mTHS and suppression of urge to smoke were comparable to mCC.ConclusionSwitching from mCCs to mTHS significantly reduced the exposure to HPHCs to levels approaching those observed in subjects who abstained from smoking for the duration of the study.ImplicationsThis study compared the impact of switching to mTHS on biomarkers of exposure, relative to continued smoking or SA.Clinical SignificanceSwitching to mTHS 2.2 led to significant reductions of exposure (total NNAL, total NNN, COHb, MHBMA, 3-HPMA, S-PMA, Total 1-OHP, 4-ABP, 1-NA, 2-NA, o-tol, CEMA, HEMA, HMPMA, and B[a]P) after 5 days in confinement, which were maintained throughout the subsequent ambulatory period of 86 days. The reductions were similar to those observed upon SA.Moderate compliance in the mTHS group during the ambulatory period was indicative of dual use of mTHS and mCC.mTHS provides an acceptable alternative to mCC for adult smokers.Trial RegistrationNCT01989156 (ClinicalTrials.gov).

3D human microvessel-on-a-chip model for studying monocyte-to-endothelium adhesion under flow - application in systems toxicology.

Lifestyle and genetic factors can lead to the development of atherosclerosis and, ultimately, cardiovascular adverse events. Rodent models are commonly used to investigate mechanism(s) of atherogenesis. However, the 3Rs principles, aiming to limit animal testing, encourage the scientific community to develop new physiologically relevant in vitro alternatives. Leveraging the 96-chip OrganoPlate®, a microfluidic platform, we have established a three-dimensional (3D) model of endothelial microvessels-on-a-chip under flow using primary human coronary arterial endothelial cells. As functional readout, we have set up an assay to measure the adhesion of monocytes to the lumen of perfused microvessels. For monitoring molecular changes in microvessels, we have established the staining and quantification of specific protein markers of inflammation and oxidative stress using high content imaging, as well as analyzed transcriptome changes using microarrays. To demonstrate its usefulness in systems toxicology, we leveraged our 3D vasculature-on-a-chip model to assess the impact of the Tobacco Heating System (THS) 2.2, a candidate modified risk tobacco product, and the 3R4F reference cigarette on the adhesion of monocytic cells to endothelial microvessels. Our results show that THS 2.2 aerosol-conditioned medium had a reduced effect on monocyte-endothelium adhesion compared with 3R4F smoke-conditioned medium. In conclusion, we have established a relevant 3D vasculature-on-a-chip model for investigating leukocyte-endothelial microvessel adhesion. A case study illustrates how the model can be used for product testing in the context of systems toxicology-based risk assessment. The current model and its potential further development options also open perspectives of applications in vascular disease research and drug discovery.

Comparison of monoamine oxidase inhibition by cigarettes and modified risk tobacco products.

The adverse effects of cigarette smoking are well documented, and the two main strategies for reducing smoking prevalence are prevention of smoking initiation and promotion of smoking cessation. More recently, a third and complementary avenue, tobacco harm reduction has emerged, which is aimed to reduce the burden of smoking-related diseases. This has been enabled by the development of novel products such as electronic cigarettes (e-cigarettes) and heated tobacco products, designed to deliver nicotine with significantly reduced levels of the toxicants that are emitted by cigarettes. Several potential modified risk tobacco products (pMRTP) have been reported to emit significantly less toxicants than cigarettes and significantly reduce toxicant exposure in smokers who switch completely to such products. These are two prerequisites for pMRTPs to reduce harm and the risk of smoking-related disease. However, concerns remain regarding the addictive nature of these products. Smoking addiction is a complex phenomenon involving multiple pharmacological and non-pharmacological factors. Although the main pharmacological substance associated with smoking addiction is nicotine, accumulating evidence suggests that nicotine mostly acts as a primary reinforcer and that other factors are involved in establishing smoking addiction. Inhibition of monoamine oxidases (MAO)-mammalian flavoenzymes with a central role in neurotransmitter metabolism-has also been suggested to be involved in this process. Therefore, we aimed to comparatively investigate the ability of several types of pMRTPs and cigarette smoke (3R4F) to inhibit MAO activity. The results showed that the heated tobacco product Tobacco Heating System (THS) 2.2 and the MESH 1.1 e-cigarette possessed no MAO inhibitory activity while 3R4F significantly inhibits the levels of MAO activity (3R4F MAO-A and B; > 2 μM nicotine). Snus products have similar inhibition profiles as 3R4F but for larger nicotine concentrations (snus MAO-A; ∼68-fold, snus MAO-B; ∼23-fold higher compared to 3R4F). These observations were confirmed by analytical datasets of potential MAO inhibitors emitted by these products. In conclusion, we have demonstrated that specific pMRTPs, namely THS 2.2 and MESH 1.1, have a significantly lower MAO-inhibitory activity than 3R4F. These findings provide a basis for further investigation of the role of MAO inhibitors in cigarette addiction as well as the implications of the findings for abuse liability of pMRTPs in comparison with cigarettes.

Mainstream smoke constituents and in vitro toxicity comparative analysis of 3R4F and 1R6F reference cigarettes.

A new Kentucky reference cigarette, 1R6F, has been manufactured to replace the depleting 3R4F reference cigarette. The 3R4F Kentucky reference cigarettes have been widely used as monitor or comparator cigarettes for mainstream smoke analysis and in vitro and in vivo toxicological data of cigarettes and novel tobacco products. Both reference cigarettes were analyzed in the same laboratory during the same period of time with the goal of performing a comparison of 3R4F and 1R6F. On the basis of the results obtained from aerosol chemistry and in vitro assays, we consider that the 1R6F reference cigarette is a suitable replacement for the 3R4F reference cigarette as a comparator/monitor cigarette. Its specific use as a comparator for novel tobacco products was checked on the basis of a comparative test with the Tobacco Heating System 2.2 as an example.

Effects of cigarette smoke and tobacco heating aerosol on color stability of dental enamel, dentin, and composite resin restorations.

To test if cigarette smoke (CS) causes discoloration of enamel, dentin, and composite resin restorations and induces color mismatch between dental hard tissues and the restorations, and to compare the findings with the effects of aerosol generated by the tobacco heating system (THS) 2.2. Twenty-two human premolars were prepared with Class V cavities restored with Filtek Supreme Ultra (3M Espe) composite resin. Teeth were divided into two groups and exposed to either CS from 20 reference cigarettes (3R4F) or aerosol from 20 THS 2.2 tobacco heat sticks 4 days a week for 3 weeks. CIE L*a*b* color was assessed before and after exposure and brushing at 1, 2, and 3 weeks. Color match, marginal discoloration, marginal integrity, and surface texture of the Class V restoration were assessed according to a modified US Public Health Service (USPHS) criterion. Marked discoloration of enamel and dentin was observed following 3 weeks of CS exposure (ΔE = 8.8 ± 2.6 and 21.3 ± 4.4, respectively), and color mismatch occurred between the composite resin restorations (ΔE = 25.6 ± 3.8) and dental hard tissues. Discoloration was minimal in the enamel, dentin, and composite resin restorations in the THS 2.2 group, and no color mismatch was observed after 3 weeks of THS 2.2 aerosol exposure. CS causes significant tooth discoloration and induces color mismatch between dental hard tissues and composite resin restorations. Reducing or eliminating the deposits derived from tobacco combustion could minimize the impact of tobacco products on tooth discoloration.

Investigation of menthol content and transfer rates in cigarettes and Tobacco Heating System 2.2

Menthol cigarettes account for a significant market share in many countries. However, little recent data exists on menthol levels in cigarettes and in mainstream smoke, limited to some markets or specific cigarette designs, such as cigarettes containing capsules filled with flavoring liquids. Samples of mentholated cigarettes bought worldwide with a variety of cigarette designs were analyzed for menthol content in cigarettes and in cigarette mainstream smoke with two analytical machine smoking regimes. A wide range of menthol content in cigarettes, from 1 to 22 mg/cigarette, was observed. The transfer of menthol to the cigarette mainstream smoke was generally slightly higher than the transfer of nicotine, with a range of 17%–40% using an intense smoking regime and 1%–17% using the ISO smoking regime. For the Tobacco Heating System (THS) 2.2, the menthol content was 12.8 mg/stick, and the transfer of menthol into the aerosol was about 17% with the ISO intense smoking regime, similar to the transfer of nicotine. The menthol content of the novel product THS 2.2 corresponds to the midpoint of the menthol content range of cigarettes, and the transfer of menthol to its aerosol is in the low range of cigarette menthol transfer.

Tobacco Heating System 2.2 has a limited impact on DNA methylation of candidate enhancers in mouse lung compared with cigarette smoke

Cigarette smoke (CS) exposure has been shown to correlate with changes in DNA methylation levels, however, the impact of CS on DNA methylation at genome-wide scale is missing. Here, we used whole-genome bisulfite sequencing to assess the effects of CS extract and aerosol from the Tobacco Heating System (THS) 2.2, a candidate modified risk tobacco product, on DNA methylation in lung and liver tissues from apolipoprotein E-deficient mice during an eight-month period of exposure. We found that in lung tissue, CS mainly induced hypermethylation of candidate enhancers at late time points, while promoters were less affected. This effect was strongly reduced upon cessation or switching to THS 2.2. By contrast, chronic exposure to THS 2.2 had a limited effect on DNA methylation at both promoters and enhancers. We also identified members of the Ets and Fox families of transcription factors as potential players in the epigenetic response to CS exposure in lung tissue. In contrast to the lung, DNA methylation in the liver was largely insensitive to all investigated exposures.In summary, our investigations indicate that CS-related DNA methylation alterations are tissue-specific, occur mainly at enhancers and are strongly reduced upon smoking cessation or switching to THS2.2.

Development and validation of a method for quantification of two tobacco-specific nitrosamines in indoor air

A sensitive and accurate method for the quantification of 1′-Demethyl-1′-nitrosonicotine (NNN) and 4-(methylnitrosamino)-1-(3-Pyridyl)-1-butanone (NNK) in indoor air was developed and validated. To this aim, a novel approach for the collection of two tobacco-specific nitrosamines, using silica sorbent cartridges followed by simplified sample preparation and isotope dilution liquid chromatography/electrospray ionization tandem mass spectrometry, was applied. This procedure led to a substantial improvement in terms of sensitivity and sample throughput as compared with methods using conventional trapping. For the validation, a matrix-based approach using an accuracy profile procedure was selected. The evaluated matrices were background air samples, environmental aerosols of a heat-not-burn tobacco product (Tobacco Heating System [THS] 2.2, commercialized under the brand IQOS®), a rechargeable electronic cigarette (Solaris®), and the environmental tobacco smoke (ETS) of a conventional cigarette (Marlboro Gold®). The method showed excellent recoveries, sensitivity, and precision. The limits of detection of the method for NNN and NNK were 0.0108 ng/m3 and 0.0136 ng/m3, respectively. The calibration range of the instrument spanned 0.2–60 ng/mL. The calculated lower working range limit (LWRL) of the method for NNN was 0.126 ng/m3, and the LWRL for NNK was 0.195 ng/m3. The method was applied to evaluate surrogate environmental aerosols generated using smoking machines. This model is reliable but gives a large overestimation of the possible impact of THS 2.2 and e-cigarettes on indoor air, because the retention of NNN and NNK in the body of the consumers is not taken into account. As a consequence, the values reported do not reflect a real-life setting. The contents of the two target compounds in the surrogate environmental aerosols were 0.0830 ± 0.0153 ng/m3 of NNN and 0.0653 ± 0.0138 ng/m3 of NNK for THS 2.2, 0.0561 ± 0.0296 ng/m3 of NNN for e-cigarettes, and 0.816 ± 0.109 ng/m3 of NNN and 4.13 ± 1.04 ng/m3 NNK for cigarettes. These values correspond to 10% of the measured ETS concentration for NNN in environmental aerosols of THS 2.2 and 7% for those of e-cigarettes. For NNK, the value for the environmental aerosol of THS 2.2 was 2% of the ETS value.

Heat-Not-Burn Tobacco Products: The Devil in Disguise or a Considerable Risk Reduction?

Background: Heat-not-burn (HNB) tobacco products are not burnt but instead are inserted into a tobacco-heating system, which heats the tobacco at temperatures below that required to initiate combustion. This mechanism potentially results in significantly reduced concentrations of heat-generated toxicants in the inhalable aerosol.Method: The margin of exposure (MOE) approach was applied for quantitative risk assessment. The MOE is defined as the ratio between the toxicological threshold and the estimated human intake of the same compound. The higher the MOE, the lower the risk of a compound.Findings: The MOEs were increased by factors of 3 to 415 for the most toxic compounds in tobacco smoke, comparing use of HNB with smoking conventional tobacco products. The combined MOE for all compounds was increased 23-fold, excluding nicotine, or 10-fold including nicotine. Thus, the overall risk for cumulative toxic effects was markedly lower for HNB products.Conclusions: HNB tobacco reduced the risk of exposure to 9 out of the 20 most toxic compounds in tobacco beyond an MOE threshold of 10,000. While our results show that use of HNB products leads to a considerable risk reduction compared to conventional tobacco, the products cannot be considered completely “risk-free” due to risk of exposure to the remaining toxicants with MOE below the threshold.

Heat-Not-Burn Tobacco Products: The Devil in Disguise or a Considerable Risk Reduction?

Background: Heat-not-burn (HNB) tobacco products are not burnt but instead are inserted into a tobacco-heating system, which heats the tobacco at temperatures below that required to initiate combustion. This mechanism potentially results in significantly reduced concentrations of heat-generated toxicants in the inhalable aerosol.Method: The margin of exposure (MOE) approach was applied for quantitative risk assessment. The MOE is defined as the ratio between the toxicological threshold and the estimated human intake of the same compound. The higher the MOE, the lower the risk of a compound.Findings: The MOEs were increased by factors of 3 to 415 for the most toxic compounds in tobacco smoke, comparing use of HNB with smoking conventional tobacco products. The combined MOE for all compounds was increased 23-fold, excluding nicotine, or 10-fold including nicotine. Thus, the overall risk for cumulative toxic effects was markedly lower for HNB products.Conclusions: HNB tobacco reduced the risk of exposure to 9 out of the 20 most toxic compounds in tobacco beyond an MOE threshold of 10,000. While our results show that use of HNB products leads to a considerable risk reduction compared to conventional tobacco, the products cannot be considered completely “risk-free” due to risk of exposure to the remaining toxicants with MOE below the threshold.

The In Vitro assessment of Respiratory Sensitisation Potential of Electronic Cigarette Liquids

Experimental studies clearly demonstrate a causal effect of cigarette smoking on cardiovascular disease. To reduce the individual risk and population harm caused by smoking, alternative products to cigarettes are being developed. We recently reported on an apolipoprotein E-deficient (Apoe−/-) mouse inhalation study that compared the effects of exposure to aerosol from a candidate modified risk tobacco product, Tobacco Heating System 2.2 (THS2.2), and smoke from the reference cigarette (3R4F) on pulmonary and vascular biology. Here, we applied a transcriptomics approach to evaluate the impact of the exposure to 3R4F smoke and THS2.2 aerosol on heart tissues from the same cohort of mice. The systems response profiles demonstrated that 3R4F smoke exposure led to time-dependent transcriptomics changes (False Discovery Rate (FDR) < 0.05; 44 differentially expressed genes at 3-months; 491 at 8-months). Analysis of differentially expressed genes in the heart tissue indicated that 3R4F exposure induced the downregulation of genes involved in cytoskeleton organization and the contractile function of the heart, notably genes that encode beta actin (Actb), actinin alpha 4 (Actn4), and filamin C (Flnc). This was accompanied by the downregulation of genes related to the inflammatory response. None of these effects were observed in the group exposed to THS2.2 aerosol.

P19-06 Retrospective histopathology evaluation of respiratory tract tissues from two rat 90-day OECD 413 inhalation studies: Tobacco Heating System 2.2 versus Mentholated Tobacco Heating System 2.2 compared with conventional and mentholated reference cigarettes

P19-06 Retrospective histopathology evaluation of respiratory tract tissues from two rat 90-day OECD 413 inhalation studies: Tobacco Heating System 2.2 versus Mentholated Tobacco Heating System 2.2 compared with conventional and mentholated reference cigarettes

Sharing in the INTERVALS platform study details and data from the assessment of the impact of aerosol from the carbon-heated tobacco product (CHTP) 1.2, a potential modified risk tobacco product, on human organotypic gingival cultures

Tobacco-specific nitrosamines (TSNA) levels in tobacco cut filler and cigarette smoke were measured in more than 1000 commercially available cigarettes sampled between 2008 and 2014. Relative contributions to their transfer from tobacco to the mainstream smoke in terms of direct transfer by distillation, pyrorelease, and pyrosynthesis were evaluated on the basis of the comparison with the transfer of nicotine from tobacco to smoke. N′-nitrosonornicotine (NNN) was transferred essentially by distillation, while N′-nitrosoanatabine (NAT), 4-(methylnitrosamino)-1-(3-bipyridyl)-1-butanone (NNK) and N′-nitrosoanabasine (NAB) were transferred by pyrorelease or pyrosynthesis as well. In the case of the Tobacco Heating System 2.2, the transfer of nicotine from tobacco to the aerosol was similar to that observed for cigarettes, while the % transfer of TSNAs from tobacco to THS 2.2 aerosol was 2–3 times lower than in cigarettes. This difference is due to the fact that the tobacco is heated instead of burnt resulting in a lower direct transfer by distillation and a lower if any contribution of pyrosynthesis or pyrorelease.

Evaluation of Biological and Functional Changes in Healthy Smokers Switching to the Tobacco Heating System 2.2 Versus Continued Tobacco Smoking: Protocol for a Randomized, Controlled, Multicenter Study.

BackgroundTobacco harm reduction, substituting less harmful tobacco products for combustible cigarettes, is a complementary approach for smokers who would otherwise continue to smoke. The Philip Morris International (PMI) Tobacco Heating System (THS) 2.2 is a novel tobacco product with the potential to reduce the risk of harm in smokers compared to continued smoking of combustible cigarettes. It heats tobacco electrically in a controlled manner, never allowing the temperature to exceed 350°C, thereby preventing the combustion process from taking place and producing substantially lower levels of toxicants while providing nicotine, taste, ritual, and a sensory experience that closely parallels combustible cigarettes. Previous clinical studies have demonstrated reduced exposure to the toxicants (approaching the levels observed after quitting) for smokers who switched to THS 2.2, for three months. For adult smokers who would otherwise continue smoking combustible cigarettes, switching to THS 2.2 may represent an alternative way to reduce the risk of tobacco-related diseases.ObjectiveThis study aimed to further substantiate the harm reduction potential of THS 2.2 by demonstrating favorable changes in a set of 8 coprimary endpoints, representative of pathomechanistic pathways (ie, inflammation, oxidative stress, lipid metabolism, respiratory function, and genotoxicity), linked to smoking-related diseases, in smokers switching from combustible cigarettes to THS 2.2.MethodsThis study was a randomized, controlled, two-arm parallel group, multicenter ambulatory US study conducted in healthy adult smokers switching from combustible cigarettes to THS 2.2 compared with smokers continuing to smoke combustible cigarettes for six months. Subjects had a smoking history of at least ten years and did not intend to quit within the next six months.ResultsEnrollment started in March 2015 and the trial was completed in September 2016. In total, 984 subjects were randomized (combustible cigarettes, n=483; THS 2.2, n=477), and 803 completed the study. The results are expected to be available in a subsequent publication in 2019.ConclusionsIn this paper, we describe the rationale and design for this clinical study that focused on the evaluation of THS 2.2’s potential to reduce the risk of smoking-related diseases compared with that of combustible cigarettes. This study will provide insights regarding favorable changes in biological and functional endpoints informed by effects known to be seen upon smoking cessation.Trial RegistrationClinicalTrials.gov NCT02396381; http://clinicaltrials.gov/ct2/show/NCT02396381 (Archived by WebCite at http://www.webcitation.org/71PCRdagP)Registered Report IdentifierRR1-10.2196/11294

Investigation and comparison of the transfer of TSNA from tobacco to cigarette mainstream smoke and to the aerosol of a heated tobacco product, THS2.2

Tobacco-specific nitrosamines (TSNA) levels in tobacco cut filler and cigarette smoke were measured in more than 1000 commercially available cigarettes sampled between 2008 and 2014. Relative contributions to their transfer from tobacco to the mainstream smoke in terms of direct transfer by distillation, pyrorelease, and pyrosynthesis were evaluated on the basis of the comparison with the transfer of nicotine from tobacco to smoke. N′-nitrosonornicotine (NNN) was transferred essentially by distillation, while N′-nitrosoanatabine (NAT), 4-(methylnitrosamino)-1-(3-bipyridyl)-1-butanone (NNK) and N′-nitrosoanabasine (NAB) were transferred by pyrorelease or pyrosynthesis as well. In the case of the Tobacco Heating System 2.2, the transfer of nicotine from tobacco to the aerosol was similar to that observed for cigarettes, while the % transfer of TSNAs from tobacco to THS 2.2 aerosol was 2–3 times lower than in cigarettes. This difference is due to the fact that the tobacco is heated instead of burnt resulting in a lower direct transfer by distillation and a lower if any contribution of pyrosynthesis or pyrorelease.

Cardiovascular Effects Observed when Using the Tobacco Heating System (THS) Compared with Continued Smoking

Cardiovascular Effects Observed when Using the Tobacco Heating System (THS) Compared with Continued Smoking

Determination of eight carbonyl compounds in aerosols trapped in phosphate buffer saline solutions to support in vitro assessment studies

When investigating the toxicological impact of aerosols using in vitro systems like cell cultures, it is essential to have a quantitative measurement of the chemicals that the cells are exposed to.Carbonyl compounds represent an important class of marker compounds for in vitro and in vivo exposure to different toxicological agents, including cigarette smoke (CS). A new LC-MS/MS method that quantifies eight of these analytes in aerosols trapped in phosphate-buffered saline solutions has been developed to measure exposure. During the method development phase, particular attention has been paid to the efficient derivatization of the target compounds in the trapped aerosols and to avoid the formation of poly-derivatized molecules, which could lead to inaccurate quantifications.The method has been successively validated using the accuracy profile procedure. Selectivity, detection limits, precision, and accuracy have been evaluated for Vitrocell®, Gas Vapor Phase (GVP), and Whole Smoke (WS) matrices of smoke generated by 3R4F cigarettes and aerosol generated by the Tobacco Heating System (THS) 2.2, a heat-not-burn tobacco product developed by Philip Morris International (Smith et al., 2016) [1]. Validation results confirmed that the established working ranges also allow the analysis of THS aerosols, where the concentrations of carbonyl compounds are substantially lower than those generated by 3R4F cigarettes. Moreover, data gathered on 3R4F aerosol samples trapped with DNPH in acetonitrile solutions have been compared to the quantification given by an in-house UHPLC-MS/MS and reference values from the literature.

Assessment of mitochondrial function following short- and long-term exposure of human bronchial epithelial cells to total particulate matter from a candidate modified-risk tobacco product and reference cigarettes

Mitochondrial dysfunction caused by cigarette smoke is involved in the oxidative stress-induced pathology of airway diseases. Reducing the levels of harmful and potentially harmful constituents by heating rather than combusting tobacco may reduce mitochondrial changes that contribute to oxidative stress and cell damage. We evaluated mitochondrial function and oxidative stress in human bronchial epithelial cells (BEAS 2B) following 1- and 12-week exposures to total particulate matter (TPM) from the aerosol of a candidate modified-risk tobacco product, the Tobacco Heating System 2.2 (THS2.2), in comparison with TPM from the 3R4F reference cigarette. After 1-week exposure, 3R4F TPM had a strong inhibitory effect on mitochondrial basal and maximal oxygen consumption rates compared to TPM from THS2.2. Alterations in oxidative phosphorylation were accompanied by increased mitochondrial superoxide levels and increased levels of oxidatively damaged proteins in cells exposed to 7.5 μg/mL of 3R4F TPM or 150 μg/mL of THS2.2 TPM, while cytosolic levels of reactive oxygen species were not affected. In contrast, the 12-week exposure indicated adaptation of BEAS-2B cells to long-term stress. Together, the findings indicate that 3R4F TPM had a stronger effect on oxidative phosphorylation, gene expression and proteins involved in oxidative stress than TPM from the candidate modified-risk tobacco product THS2.2.