Fowler Kathy, Fields Wanda, Hargreaves Victoria, Reeve Lesley, Bombick Betsy
RAI Services Company, Scientific & Regulatory Affairs, 401 North Main Street, Winston-Salem, NC 27101, USA.
Covance Laboratories Ltd., North Yorkshire, UK.
Toxicol Rep. 2018 Apr 12;5:542-551. doi: 10.1016/j.toxrep.2018.04.003. eCollection 2018.
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 VITROCELLVC10 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 -like conditions for assessing the toxicological impact of fresh WS in assays. The goals of this study were to 1) qualify the VITROCELLVC10 to demonstrate functionality of the system, 2) develop and validate the Ames test following WS exposure with the VITROCELLVC10 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 VITROCELLVC10 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 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.
艾姆斯试验已被用于评估烟草产品的潜在致突变性，但通常是使用分离暴露（如总颗粒物[TPM]、气相[GVP]）而非全烟（WS）进行。VITROCELL VC10烟雾暴露系统提供了多个平台，用于气液界面（ALI），在艾姆斯试验暴露的情况下则是气-琼脂界面（AAI），以模拟评估新鲜全烟在试验中的毒理学影响的类似条件。本研究的目标是：1）鉴定VITROCELL VC10以证明系统的功能；2）开发并验证使用VITROCELL VC10进行全烟暴露后的艾姆斯试验；3）评估艾姆斯试验区分参考可燃产品（3R4F肯塔基参考卷烟）和主要烟草加热产品（Eclipse）的能力。基于关键功能评估，证明VITROCELL VC10适合持续产生全烟的目的。对5种细菌菌株（TA97、TA98、TA100、TA1535和TA102）进行了试验验证，在暴露于3R4F全烟后，在存在大鼠肝脏S-9的情况下，观察到TA98和TA100中回复突变体有与暴露相关的可重复变化。在比较研究中，观察到在存在S9的情况下，TA98和TA100暴露于3R4F和Eclipse全烟后，致突变性有与暴露相关的变化，Eclipse的反应明显小于3R4F（p＜0.001），这与主要加热该产品释放的化学成分较少一致。
