Background and Aim: Toothbrushing can potentially abrade the teeth and direct and indirect dental restorations, and may also affect their color. This study aimed to assess the color change (Δ, E) of Ceramill Zolid FX following abrasion with/without toothpaste in two groups of simultaneous and separate staining and glazing. Materials and Methods: This in vitro, experimental study was conducted on 40 cylindrical specimens measuring 2 x 10 mm fabricated from Ceramill Zolid FX by the computer-aided design/computer-aided manufacturing technology. The samples were randomly divided into four groups (simultaneous and separate staining and glazing) and subjected to simulation of 3, 6 and 9 years of toothbrushing in a cross-brushing machine with and without toothpaste. The Δ, E of specimens was measured before and after the intervention by using Shade Pilot spectrophotometer, and compared using the Mann-Whitney and Friedman tests (alpha=0. 05). Results: Significant differences were noted in Δ, E of simultaneous staining and glazing control (without toothpaste) and intervention (with toothpaste) groups over time (both P=0. 000). Both the control and intervention subgroups in separate staining and glazing group also showed significant differences in Δ, E over time (both P=0. 000). Conclusion: The results showed greater color change over time in simultaneous staining and glazing group due to the presence of one layer of stain and glaze, compared with separate staining and glazing. The effect of toothpaste on color change was only significant at 9 years in both simultaneous and separate staining and glazing groups.