In vitro permeation of nicotine and tobacco specific nitrosamines from smokeless tobacco product extracts in a 3D buccal tissue model

Tobacco product use is a risk factor in the development of oral cancer, although epidemiology studies show this risk is far less with smokeless tobacco product use than cigarette smoking. While smokeless tobacco contains harmful and potentially harmful constituents (HPHCs), the oral permeation of HPHCs in oral tobacco products is not completely understood. To improve the understanding, three different extract concentrations of the CORESTA reference products (CRP) for snus (CRP1.1) and moist snuff (CRP2.1) were applied to cellular tissue derived from two donors of EpiOral™ model, a 3D human buccal model, and permeation of nicotine and tobacco-specific nitrosamines (TSNAs) were measured over two hours. Permeation of 0.15% caffeine in complete artificial saliva and cell viability were also measured. Results showed that a consistent and concentration dependent cumulative permeation of nicotine and TSNAs was observed with high percent recovery in all conditions. A high degree of sensitivity was seen for all analytes, with minimal cytotoxicity for both CRPs. The data presented here show the EpiOral™ model is fit-for-purpose to evaluate the permeation of nicotine and TSNAs in nicotine-containing snus and moist snuff oral tobacco.

EpiOral (ORL-200, ORL-200-D2), TEER, MTT, permeation, percent recovery, nicotine, tobacco specific nitrosamines, TSNA, smokeless tobacco, CORESTA reference products, snus, CRP1.1, moist snuff, CRP2.1, caffeine, complete artificial saliva, harmful constituents, 4-(methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK), benzo[a] pyrene (B[a]P), nitroso-nornicotine (NNN), nitrosoanatabine (NAT), nitroso-anabasine (NAB)

CORESTA reference products, snus, CRP1.1, moist snuff, CRP2.1