Aim: To investigate the effects of selected alcohol-free mouthwashes with different formulations (zinc-hydroxyapatite, hydrogen peroxide, and sodium fluoride) on the surface microhardness of a single-shade universal resin composite. Materials and Methods: Forty disc-shaped specimens (8 × 2 mm) from the universal resin composite (Omnichroma®), and a nano-hybrid composite (Tetric® N-Ceram) were prepared. After polymerization, baseline surface microhardness values were recorded using Vickers microhardness tester. The samples from each material were randomly assigned to 4 groups ( n = 10) and immersed in 20 ml of the mouthwashes: Biorepair®, Listerine®, Colgate® Optic White, and distilled water (control). The samples were kept in the immersion solutions for 24 hours, and post-immersion microhardness values were recorded. Data were analyzed with one-way ANOVA and paired sample t-tests at p < .05. Results: Significant reduction in microhardness was observed in all resin composite groups after immersion in the mouthwashes compared to baseline values ( p < .0001). The highest microhardness reduction in Omnichroma® group was observed after immersion in Colgate® Optic White; and Tetric® N-Ceram group after immersion in Listerine® mouthwash. For both materials, the least reduction in microhardness was observed after immersion in Biorepair®. Microhardness values for Omnichroma were significantly higher than Tetric® N-Ceram ( p < .0001). However, Omnichroma exhibited a significantly greater reduction in microhardness after immersion in the tested mouthwashes. Conclusion: In vitro simulated use of the investigated mouthwashes negatively affected the surface microhardness of both tested resin composites. The observed effects were both mouthwash and material dependent.