Electronic cigarettes, also known as ecig-s, have been widely used in recent years, especially among younger populations. Cytotoxic and carcinogenic substances can be present in their composition, but little is known about the risks associated with their use. The purpose of this study was to evaluate the proteome profile of saliva from users of electronic cigarettes. Participants were divided into two groups: the Electronic Cigarette Group (EG), consisting of 25 regular and exclusive e-cig users, and the Control Group (CG), comprising 25 non-smokers and non-e-cig users, matched by sex and age to the EG. All participants underwent clinical examination and unstimulated saliva collection for evaluation of the salivary proteome. Shotgun proteomics resulted in the identification of 168 proteins, including 48 exclusive to EG, 28 exclusive to CG, and 92 shared by both groups. Statistical comparison (t-test) between EG and CG revealed four differentially expressed proteins: Fatty acid binding protein 5, Calgranulin-B, and Cornifin-A in the CG, and cytostatin-D in the EG. Additionally, a higher abundance of proteins was observed, including glutathione S-transferase P, Immunoglobulin J chain and Acetyl-CoA dehydrogenase, in the EG group. These findings suggest an organismal response to increased oxidative stress, the presence of inflammatory components, and altered salivary microbiome, which can predispose individuals to the development of future pathological conditions.