Differential Expression of Salivary Biomarkers in Smokeless Tobacco Users: A Cross‐Sectional Study

OBJECTIVE: The use of smokeless tobacco (SLT) is associated with significant harm to oral health and salivary biomarkers have recently emerged as potential indicators of such harm. This study evaluated levels of key salivary proteins, including inflammatory cytokines and vascular dysfunction biomarkers, among SLT users and examined the influence of usage characteristics on these levels. METHODS: A total of 62 male participants, aged 24-55 years, were equally allocated to either SLT users or controls (non-users) groups. Expression of salivary interleukin 1β (IL-1β), tumor necrosis factor alpha (TNF-α), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), endothelin-1 (ET1), and transferrin (TF) was evaluated using ELISA. Multiple linear regression assessed effects of mixed SLT use, areca-nut content, duration and frequency of use on biomarker expression. RESULTS: SLT users exhibited significantly elevated levels of VEGF, EGF, and ET1 (p < 0.0001), TF, and IL-1β (p < 0.05) compared to controls. Additionally, using more than one type of SLT was significantly associated with higher expression of VEGF and TF. CONCLUSIONS: SLT consumption is associated with significant upregulation of pro-inflammatory cytokines and biomarkers of endothelial dysfunction, while mixed use of SLT products may further potentiate these effects. These findings highlight the potential pathogenic impact of SLT on oral health.