Advanced in vitro exposure systems.

Development, qualification, validation and application of the Ames test using a VITROCELL® VC10® smoke exposure system

12. Apr. 2018

https://doi.org/10.1016/j.toxrep.2018.04.003

Kathy Fowlera, Wanda Fieldsa, Victoria Hargreavesb, Lesley Reeveb, Betsy Bombicka,
a RAI Services Company, Scientific & Regulatory Affairs, 401 North Main Street, Winston-Salem, NC 27101, USA2
b Covance Laboratories Ltd., North Yorkshire, UK

The article shows the successfull qualification of the Vitrocell VC10 smoking robot. The robot was used to  expose bacterial cells to cigarette whole smoke generated from 3R4F reference cigarettes and Eclipse cigarettes. A comparison was made between whole smoke from a tobacco heating and a combustible cigarette.

 

ABSTRACT
The Ames test has established use in the assessment of potential mutagenicity of tobacco products but has generally been performed using partitioned exposures (e.g. total particulate matter [TPM], gas vapor phase [GVP]) rather than whole smoke (WS). The VITROCELL®VC10® smoke exposure system offers multiple platforms for air liquid interface (ALI), or air agar interface (AAI) in the case of the Ames test exposure to mimic in vivo-like conditions for assessing the toxicological impact of fresh WS in in vitro assays. The goals of this study were to 
1) qualify the VITROCELL®VC10® to demonstrate functionality of the system,
2) develop and validate the Ames test following WS exposure with the VITROCELL®VC10® and 
3) assess the ability of the Ames test to differentiate between a reference combustible product (3R4F Kentucky reference cigarette) and a primarily tobacco heating product (Eclipse). Based on critical function assessments, the VITROCELL ®VC10® was demonstrated to be fit for the purpose of consistent generation of WS. Assay validation was conducted for 5 bacterial strains (TA97, TA98, TA100, TA1535 and TA102) and reproducible exposure–related changes in revertants were observed for TA98 and TA100 in the presence of rat liver S-9 following exposure to 3R4F WS. In the comparative studies, exposure-related changes in in vitro mutagenicity following exposure of TA98 and TA100 in the presence of S9 to both 3R4F and Eclipse WS were observed, with the response for Eclipse being significantly less than that for 3R4F (p < 0.001) which is consistent with the fewer chemical constituents liberated by primarily-heating the product.

 

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