Objective: The use of electronic cigarettes (ECIGs) has recently increased as an alternative to conventional smoking tobacco products. The literature is scarce on the effects of ECIGs on dental and oral structures. Therefore, the objective of this study was to observe whether ECIG aerosol could alter the color of dental enamel. Materials and methods: Sixty-three bovine enamel specimens were randomly separated into groups (n = 7) and treated with aerosols with different e-liquid flavors (neutral, menthol, and tobacco) and nicotine content (0, 12, and 18 mg). The initial color assessment was performed using a spectrophotometer (Easy Shade—Vita). Dental enamel was exposed to 20 cycles of ECIG aerosol in a smoking machine, and the final color was measured. The color change was evaluated using ΔEab, ΔE00, and Whiteness Index (WI D ) formulae. Differences in L* a* b* coordinates were calculated, and data were analyzed (two-way ANOVA, uncorrected Fisher's LSD post hoc test, α = 0.05). Results: Luminosity (ΔL) was reduced by aerosols with almost all levels of nicotine content and neutral and menthol flavors. Flavored e-liquids caused more color change (P <.05) according to ΔEab and ΔE00. ΔE values from both formulae exceeded the thresholds for perceptible visual alterations of color. WI D increased after ECIGs exposure for menthol and tobacco and decreased for neutral flavors. Conclusions: ECIG aerosol from e-liquids with different nicotine contents and flavors altered enamel color. Menthol and tobacco e-liquids may alter the enamel color decreasing the yellowness of the enamel compared to neutral e-liquid. Clinical significance: Electronic cigarettes can cause perceptible changes in tooth color, altering dental esthetics.