Assessment of cigarette smoke particle deposition within the Vitrocell® exposure module using quartz crystal microbalances

Adamson, Jason; Thorne, David; Dalrymple, Annette; Dillon, Debbie; Meredith, Clive

doi:10.1186/1752-153x-7-50

Cited by 41 publications

(47 citation statements)

References 35 publications

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“…QCM technology has previously been described in a set-up similar to this by Adamson et al, 2013 [17] and has been shown to correlate with particulate spectrofluorescence techniques. During the whole smoke generation and exposure phase, the QCM took mass readings every 2 seconds in real-time.…”

Section: Methodsmentioning

confidence: 99%

Development of a BALB/c 3T3 neutral red uptake cytotoxicity test using a mainstream cigarette smoke exposure system

et al. 2014

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BackgroundTobacco smoke toxicity has traditionally been assessed using the particulate fraction under submerged culture conditions which omits the vapour phase elements from any subsequent analysis. Therefore, methodologies that assess the full interactions and complexities of tobacco smoke are required. Here we describe the adaption of a modified BALB/c 3T3 neutral red uptake (NRU) cytotoxicity test methodology, which is based on the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) protocol for in vitro acute toxicity testing. The methodology described takes into account the synergies of both the particulate and vapour phase of tobacco smoke. This is of particular importance as both phases have been independently shown to induce in vitro cellular cytotoxicity.FindingsThe findings from this study indicate that mainstream tobacco smoke and the gas vapour phase (GVP), generated using the Vitrocell® VC 10 smoke exposure system, have distinct and significantly different toxicity profiles. Within the system tested, mainstream tobacco smoke produced a dilution IC50 (dilution (L/min) at which 50% cytotoxicity is observed) of 6.02 L/min, whereas the GVP produced a dilution IC50 of 3.20 L/min. In addition, we also demonstrated significant dose-for-dose differences between mainstream cigarette smoke and the GVP fraction (P < 0.05). This demonstrates the importance of testing the entire tobacco smoke aerosol and not just the particulate fraction, as has been the historical preference.ConclusionsWe have adapted the NRU methodology based on the ICCVAM protocol to capture the full interactions and complexities of tobacco smoke. This methodology could also be used to assess the performance of traditional cigarettes, blend and filter technologies, tobacco smoke fractions and individual test aerosols.

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Section: Methodsmentioning

confidence: 99%

Development of a BALB/c 3T3 neutral red uptake cytotoxicity test using a mainstream cigarette smoke exposure system

et al. 2014

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“…QCM technology has been incorporated into a variety of exposure chambers [20,24,26] and has been shown to correlate strongly with particulate spectrofluorescence techniques [20]. Prior to smoke exposure, the QCM module was acclimatised for several minutes prior to the baseline being set to zero.…”

Section: Methodsmentioning

confidence: 99%

Characterisation of a Vitrocell® VC 10 in vitrosmoke exposure system using dose tools and biological analysis

Thorne

Kilford

Payne

et al. 2013

Chemistry Central Journal

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BackgroundThe development of whole smoke exposure systems have been driven by the fact that traditional smoke exposure techniques are based on the particulate phase of tobacco smoke and not the complete smoke aerosol. To overcome these challenges in this study, we used a Vitrocell® VC 10 whole smoke exposure system. For characterisation purposes, we determined smoke deposition in relationship to airflow (L/min), regional smoke deposition within the linear exposure module, vapour phase dilution using a known smoke marker (carbon monoxide) and finally assessed biological responses using two independent biological systems, the Ames and Neutral Red uptake (NRU) assay.ResultsSmoke dilution correlates with particulate deposition (R2 = 0.97) and CO concentration (R2 = 0.98). Regional deposition analysis within the linear exposure chamber showed no statistical difference in deposited mass across the chamber at any airflows tested. Biological analysis showed consistent responses and positive correlations with deposited mass for both the Ames (R2 = 0.76) and NRU (R2 = 0.84) assays.ConclusionsWe conclude that in our study, under the experimental conditions tested, the VC 10 can produce stable tobacco smoke dilutions, as demonstrated by particulate deposition, measured vapour phase smoke marker delivery and biological responses from two independent in vitro test systems.

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“…A quartz crystal microbalance (QCM) (Vitrocell ® Systems, Waldkirch, Germany) was included in situ of exposure at position 4 within the Ames module, distal to the smoke inlet, as previously demonstrated [38][39][40]. QCMs provide a gravimetric measure of deposited mass, providing a valuable quality control (QC) measure of smoke exposure within and between runs, and allows data to be presented as a function of deposited mass (g/cm 2 ).…”

Section: Measuring Dosementioning

confidence: 99%

The mutagenic assessment of mainstream cigarette smoke using the Ames assay: A multi-strain approach

Thorne

Kilford

Hollings

et al. 2015

Mutation Research/Genetic Toxicology and Environmental Mutagene

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Salmonella typhimurium strains TA1535, TA1537, TA97, TA102 and TA104 were assessed for their suitability and use in conjunction with a Vitrocell(®) VC 10 Smoking Robot and 3R4F reference mainstream cigarette smoke. Little information exists on TA97, TA104, TA1535, TA1537 and TA102 using an aerosol 35mm spread-plate format. In this study, TA1535 and TA1537 were considered sub-optimal for use with a scaled-down format, due to low spontaneous revertant numbers (0-5 revertants/plate). In the context of a regulatory environment, TA97 is deemed an acceptable alternative for TA1537 and was therefore selected for whole smoke exposure in this study. However, there is no acceptable alternative for TA1535, therefore this strain was included for whole smoke exposure. TA1535, TA97, TA102 and TA104 were assessed for mutagenic responses following exposure to cigarette smoke at varying concentrations (using diluting airflow rates of 1.0, 4.0, 8.0 and 12.0L/min), and exposure times of 24 and 64min. A positive mutagenic response to cigarette smoke was observed in strain TA104 at both the 24 and 64min time points, in the presence of S-9, at the highest smoke concentration tested (1.0L/min diluting airflow). The three remaining strains were found to be unresponsive to cigarette smoke at all concentrations tested, in the presence and absence of metabolic activation. Cigarette smoke particulate deposition was quantified in situ of exposure using quartz crystal microbalance technology, enabling data to be presented against an associated gravimetric mass (μg/cm(2)). Finally, data obtained in this study were combined with previously published Ames data for TA98, TA100, YG1024, YG1042 and Escherichia coli (WP2 uvrA pKM101), generated using the same 35mm methodology. The combined data-set was used to propose an aerosol testing strategy, based on strain compatibility with the whole smoke aerosol, whilst maintaining the essence of the regulatory guidelines for the standard Ames assay.

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Assessment of cigarette smoke particle deposition within the Vitrocell® exposure module using quartz crystal microbalances

Cited by 41 publications

References 35 publications

Development of a BALB/c 3T3 neutral red uptake cytotoxicity test using a mainstream cigarette smoke exposure system

Development of a BALB/c 3T3 neutral red uptake cytotoxicity test using a mainstream cigarette smoke exposure system

Characterisation of a Vitrocell® VC 10 in vitrosmoke exposure system using dose tools and biological analysis

The mutagenic assessment of mainstream cigarette smoke using the Ames assay: A multi-strain approach

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