In vitro evaluation of mutagenic, cytotoxic, genotoxic and oral irritation potential of nicotine pouch products

Jacqueline Miller-Holt; Irene Baskerville-Abraham; Masanori Sakimura; Toshiro Fukushima; Andrea Puglisi; Jeremie Gafner

doi:10.1016/j.toxrep.2022.06.008

In vitro evaluation of mutagenic, cytotoxic, genotoxic and oral irritation potential of nicotine pouch products

Toxicol Rep. 2022 Jun 15:9:1316-1324. doi: 10.1016/j.toxrep.2022.06.008. eCollection 2022.

Authors

Jacqueline Miller-Holt¹, Irene Baskerville-Abraham¹, Masanori Sakimura¹, Toshiro Fukushima², Andrea Puglisi¹, Jeremie Gafner¹

Affiliations

¹ Scientific and Regulatory Affairs, JT International SA, Geneva, Switzerland.
² Scientific Product Assessment Center, R&D Group, Japan Tobacco Inc., Kanagawa, Japan.

Abstract

Non-clinical in vitro studies were conducted to investigate the characteristics of extracts from tobacco free nicotine pouches alongside a reference snus product and/or 1R6F reference cigarette. In vitro investigations were conducted in the Neutral Red Uptake (NRU) cytotoxicity assay, Bacterial Reverse Mutation (Ames) assay, and in vitro Mammalian Cell Micronucleus (ivMN) assay. These products were also investigated for their oral irritation potential in the EpiGingival™ 3D tissue model. Results from the Ames, in vitro Micronucleus and NRU assays indicated that the tested products were non-mutagenic, non-genotoxic and non-cytotoxic in contrast to results obtained for the 1R6F reference cigarette. Results from Complete Artificial Saliva (CAS) extracts from these products also failed to be classified as irritants (as measured using the MTT assay), in the EpiGingival™ 3D tissue model.

Keywords: 3D tissue; Ames; Cytotoxicity; EpiGingival™; In vitro; Micronucleus; Nicotine pouch; Smokeless.