Purpose: This study aims to evaluate the effect of erosive, abrasive, and erosive/ abrasive challenges on the glaze layer of ceramic materials. Methods: Ninety-five samples of monolithic zirconia (MZ) (LuxaCam Zircon HT-Plus) and lithium disilicate (LD) (IPS e.max CAD) were divided according to the response variables: Surface roughness and surface loss (n = 10), evaluated with optical profilometry; surface topography, with scanning electron microscopy SEM (n = 3); and biofilm deposition, with microbiological assay (n = 5). The evaluations were performed in three different time evaluations: (a) Sintered, (b) Glaze, and (c) Challenge (Erosion, Abrasion, and Erosion/Abrasion). Erosion consisted in immersing specimens in HCl solution, abrasion was performed with brushing machine, and erosion/abrasion consisted of a combination of the two previous protocols. Data were analyzed with parametric tests (P < 0.05). Results: MZ glaze layer presented significantly higher surface roughness (P = 0.00), surface loss (P = 0.03), and biofilm deposition (P = 0.00) than LD. Abrasion and erosion/abrasion showed similar outcomes, generating significantly higher surface roughness (P = 0.00), surface loss (P = 0.00), and biofilm deposition (P = 0.01) than erosion. Conclusions: Glaze layer properties were altered by the challenges, with abrasion and erosion/abrasion generating higher surface roughness, surface loss, and biofilm deposition than erosion. A significant correlation was found between the surface roughness and biofilm deposition. Clinical significance: The glaze layer is susceptible to challenges, especially to abrasion and erosion/abrasion, which generated greater surface roughness and surface loss than erosion. The greater surface roughness lead to a greater biofilm deposition on the glaze layer.