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

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 [Display omitted] •Non-clinical in vitro studies on tobacco free nicotine pouch extracts.•Extracts were non-mutagenic, non-genotoxic and non-cytotoxic in contrast to results from the 1R6F reference cigarette.•Extracts were not classified as irritant (as measured using the MTT assay), in human 3D gingival epithelial cell cultures.•To our knowledge, this is the first study to assess the impact of nicotine pouch extracts on gingival organotypic cultures.