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使用VITROCELL® VC10®烟雾暴露系统在中国仓鼠卵巢（CHO）细胞中进行中性红摄取试验的开发、鉴定、验证及应用。

Development, qualification, validation and application of the neutral red uptake assay in Chinese Hamster Ovary (CHO) cells using a VITROCELL® VC10® smoke exposure system.

作者信息

Fields Wanda, Fowler Kathy, Hargreaves Victoria, Reeve Lesley, Bombick Betsy

机构信息

RAI Services Company, Scientific & Regulatory Affairs, 401 North Main Street, Winston-Salem, NC 27101, USA.

RAI Services Company, Scientific & Regulatory Affairs, 401 North Main Street, Winston-Salem, NC 27101, USA.

出版信息

Toxicol In Vitro. 2017 Apr;40:144-152. doi: 10.1016/j.tiv.2017.01.001. Epub 2017 Jan 3.

DOI:10.1016/j.tiv.2017.01.001
PMID:28062357
Abstract

UNLABELLED

Cytotoxicity assessment of combustible tobacco products by neutral red uptake (NRU) has historically used total particulate matter (TPM) or solvent captured gas vapor phase (GVP), rather than fresh whole smoke. Here, the development, validation and application of the NRU assay in Chinese Hamster Ovary (CHO) cells, following exposure to fresh whole smoke generated with the VITROCELL® VC10® system is described. Whole smoke exposure is particularly important as both particulate and vapor phases of tobacco smoke show cytotoxicity in vitro. The VITROCELL® VC10® system provides exposure at the air liquid interface (ALI) to mimic in vivo conditions for assessing the toxicological impact of smoke in vitro. Instrument and assay validations are crucial for comparative analyses.

GOALS OF THIS STUDY

  1. demonstrate functionality of the VITROCELL® VC10® system by installation, operational and performance qualification, 2) develop and validate a cellular system for assessing cytotoxicity following whole smoke exposure and 3) assess the whole smoke NRU assay sensitivity for statistical differentiation between a reference combustible cigarette (3R4F) and a primarily "heat-not-burn" cigarette (Eclipse).

RESULTS

The VITROCELL® VC10® provided consistent generation and delivery of whole smoke; exposure-related changes in in vitro cytotoxicity were observed with reproducible IC values; comparative analysis showed that the heat-not-burn cigarette was significantly (P<0.001) less cytotoxic than the 3R4F combustible cigarette, consistent with the lower levels of chemical constituents liberated by primarily-heating the cigarette versus burning.

摘要

未标注

通过中性红摄取法（NRU）对可燃烟草制品进行细胞毒性评估，历来使用的是总颗粒物（TPM）或溶剂捕获的气相（GVP），而非新鲜的全烟。在此，描述了在使用VITROCELL® VC10®系统产生的新鲜全烟暴露后，NRU检测法在中国仓鼠卵巢（CHO）细胞中的开发、验证及应用。全烟暴露尤为重要，因为烟草烟雾的颗粒相和气相在体外均显示出细胞毒性。VITROCELL® VC10®系统在气液界面（ALI）提供暴露，以模拟体内条件，用于体外评估烟雾的毒理学影响。仪器和检测验证对于比较分析至关重要。

本研究的目标

1）通过安装、操作和性能鉴定证明VITROCELL® VC10®系统的功能，2）开发并验证一种用于评估全烟暴露后细胞毒性的细胞系统，以及3）评估全烟NRU检测法对参考可燃卷烟（3R4F）和主要“加热不燃烧”卷烟（Eclipse）之间进行统计区分的灵敏度。

结果

VITROCELL® VC10®系统能持续产生并输送全烟；观察到与暴露相关的体外细胞毒性变化，且IC值具有可重复性；比较分析表明，加热不燃烧卷烟的细胞毒性显著低于3R4F可燃卷烟（P<0.001），这与主要加热卷烟而非燃烧所释放的化学成分含量较低一致。

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