Objectives The aim was to investigate color, gloss, or roughness of splint materials after storage in liquids and toothbrush simulation. Materials and methods A total of 58 x 8 (n =10 per material and group) specimens were fabricated (hand-cast, thermoforming, CAD/CAM-milled, 3D-printed materials); stored in air, water, coffee, red wine, and cleaning tablets; and investigated after fabrication, 24 h, two-, and four-week storage or toothbrushing. Color values (L*, a*, b*; ISO 11664-4:2008; CM3500d, Konica-Minolta), gloss (ISO 2813:2014), and roughness values were determined (3D laser-scanning-microscope, KJ 3D, Keyence) before and after simulation or storage. Statistics: Levene-test, one-way ANOVA, Bonferroni post hoc test, between-subjects effects, Pearson correlation (alpha = 0.05). Results Color, gloss, and roughness altered due to contact with staining solutions/toothbrush simulation. Highest impact on color, gloss, and roughness presented the material followed by storage time (Delta E material (eta 2 = 0.239/p < 0.001), storage time (eta 2 = 0.179/p < 0.001); gloss (eta 2 = 0.751/p < 0.001) (eta 2 = 0.401/p < 0.001); Ra/Rz (eta 2 >= 0.801/p < 0.001) (eta 2 >= 0.416/p <0.001)). Correlations were found between Rz and Ra (Pearson 0.887/p <= 0.001) or Rz and Delta E (0.517/p <= 0.001) or Ra and Delta E (0.460/p <= 0.001). Conclusions Storage and toothbrushing were accompanied by a change in color, gloss, and roughness. Almost all materials showed visible discoloration after 4 weeks of storage. Gloss values decreased as storage time increased. The initial roughness and polishability were better with harder materials.