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

Introduction:
This text provides an analysis of a study investigating the in vitro characteristics of extracts from tobacco-free nicotine pouch products, a reference snus product, and a 1R6F reference cigarette. The study employed various assays to evaluate the mutagenic, cytotoxic, genotoxic, and oral irritation potential of these products.

Key Points:

* The study included three in vitro assays: the Neutral Red Uptake (NRU) cytotoxicity assay, the Bacterial Reverse Mutation (ames) assay, and the in vitro Mammalian Cell Micronucleus (ivMN) assay.
* Complete artificial Saliva (CaS) extracts from the products were also tested for oral irritation potential using 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 the 1R6F reference cigarette.
* CaS extracts from these products also failed to be classified as irritants in the EpiGingival ™ 3D tissue model as measured by the MTT assay.
* The study used an independent contract research organization in Canada for the ames, NRU, and ivMN assays and Charles River Laboratories in Skokie, Illinois, USa for the EpiGingival ™ 3D tissue model testing.
* The nicotine pouches were commercially available at the time of the study, and the reference products were provided by the testing laboratories.
* The condensate samples for the ames, NRU, and ivMN assays were collected according to ISO 3308, while the condensate samples for the 3D tissue testing were collected according to the health Canada Intense hCI regime defined by ISO 20778.

Main Message:
The main message of the text is that tobacco-free nicotine pouches are non-mutagenic, non-genotoxic, and non-cytotoxic, and their CaS extracts do not irritate the oral mucosa. These findings suggest that tobacco-free nicotine pouches may offer reduced harm potential compared to traditional tobacco products. however, further research is needed to confirm these results and establish the long-term health effects of using these products. Overall, the study supports the use of in vitro assays and the EpiGingival ™ 3D tissue model as valuable tools for evaluating the safety and potential harm of novel tobacco products.

Miller-holt, J., Baskerville-abraham, I., Sakimura, M., Fukushima, T., Puglisi, a., & Gafner, J. (2022). In vitro evaluation of mutagenic, cytotoxic, genotoxic and oral irritation potential of nicotine pouch products. Toxicology Reports, 9, 1316–1324. https://doi.org/10.1016/j.toxrep.2022.06.008