Evaluation of waterpipe smoke toxicity in C57BL/6 mice model

Waterpipe smoking is a popular pastime worldwide with statistics pointing to an alarming increase in consumption. In the current paper, the evaluation of sub-chronic waterpipe smoke exposure was undertaken using C57BL/6 female mice using a dynamic exposure setting to emulate smoke exposure. Mice were daily subjected to either one (single exposure, SE) or two sessions (double exposure, DE) of waterpipe-generated smoke (two-apple flavor) for a period of two months. Although lungs histopathological examination pointed to a minor inflammation in smoke-exposed mice compared to control air-exposed (CON) group, the lung weights of the waterpipe-exposed mice were significantly higher (+72% in SE and +39% in DE) (p < 0.01) when compared to CON group. Moreover, changes in the protein expression of several proteins such as iNOS and JNK were noted in the lungs of smoke-exposed mice. However, no changes in p38 and EGFR protein levels were noted between the three groups of mice. Our results mainly showed a significant increase in urea serum levels (+28%) in SE mice along with renal pathological damage in both SE and DE mice compared to CON. Additionally, severe significant DNA damages (p < 0.05) were reported in the lungs, kidneys, bone marrow and liver of waterpipe-exposed animals, using MTS and COMET assays. These findings highlighted the significant risks posed by sub-chronic waterpipe smoke exposure in the selected animal model and the pressing need for future better management of waterpipe indoor consumption.