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The Effect of Iqmik‐derived Metals on ROS Production in Oral Epithelial Cells

Smokeless tobacco (SLT) users have 4 ‐ 6 times higher risk for developing oral cancer than non‐users. The percentage of Alaska Natives (AN) who use SLT is 4 times than that of other Alaskans. Out of all adult AN who use smokeless SLT, 35% use only iqmik, a unique combination of tobacco leaves mixed with punk fungus ash. Tobacco has a large number of carcinogens including toxic metals. The ash component of iqmik contains higher levels of pro‐inflammatory toxic metals, such as cadmium (Cd), cobalt (Co) and nickel (Ni), than tobacco. These metals trigger the generation of reactive oxygen species (ROS). The purpose of this research was to determine the intracellular redox response of primary human gingival epithelial cells (HGEP) to artificial saliva extracts of iqmik, punk ash and tobacco. Levels of Cd, Co and Ni were analyzed by ICP‐MS in iqmik, tobacco and punk ash samples and in artificial saliva extracts of iqmik, tobacco and punk ash samples following microwave‐assisted acid digestion. Intracellular redox status was quantified using H2DCFDA in primary HGEP after exposure to iqmik, tobacco and punk ash artificial saliva extracts, CdCl2, CoCl2 and NiCl2. Significantly higher concentrations of Cd, Co and Ni were isolated in punk ash and iqmik samples than in tobacco samples. Artificial saliva extracts of iqmik produced a greater than additive ROS response compared to extracts of tobacco and punk ash. Treatment with Coproduced significantly greater ROS than Cd or Ni treatment, indicating that Co may account for more of the overall ROS.