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

BMC Research Notes

Deborah Dillon¹  Clive Meredith¹  Annette Dalrymple¹  Linsey Haswell¹  Rebecca Payne²  Joanne Kilford²  David Thorne¹ 

[1] British American Tobacco, Group R&D, Southampton, Hampshire SO15 8TL, UK;Covance Laboratories Ltd, Otley Road, Harrogate, North Yorkshire HG3 1PY, UK

关键词: BALB/c3T3;    Neutral red;    Smoke exposure system;    Gas vapour phase;    Whole smoke;    Tobacco smoke;   

Others  :  1132475
DOI  :  10.1186/1756-0500-7-367

 received in 2013-11-28, accepted in 2014-06-12,  发布年份 2014

【 摘 要 】

Background

Tobacco 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.

Findings

The 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.

Conclusions

We 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.

【 授权许可】

   
2014 Thorne et al.; licensee BioMed Central Ltd.

【 图 表 】

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