Introduction: Erosion is an important cause of tooth mineral loss. The combined use of lasers and fluoride has been introduced as a novel modality for the prevention of enamel demineralization. This study aimed to assess the effect of Er:YAG laser combined with fluoride application on primary and permanent enamel resistance to erosion. Methods: Eighty enamel specimens of permanent (n=40) and primary (n=40) molars were prepared and randomly assigned to 4 groups: C-control (no pretreatment), F-acidulated phosphate fluoride (APF) gel, FL-APF gel application followed by Er:YAG laser irradiation, and LF-Er:YAG laser irradiation followed by the application of APF gel . The specimens were then submitted to pH cycling using Coca-Cola (pH = 2.4). Enamel micro-hardness was measured using the Vickers microhardness tester before pretreatment and after the erosive process. The collected data were analyzed using the Kolmogorov-Smirnov test, two-way ANOVA and repeated measures ANOVA. Results: The micro-hardness of both permanent and primary enamel specimens significantly decreased after the erosive process (P < 0.05). In the permanent enamel specimens, the greatest reduction in micro-hardness was noted in groups C and F, while the least reduction was noted in group FL. However, these differences were not statistically significant (P > 0.05). In the primary enamel specimens, the greatest reduction in micro-hardness was noted in groups C and LF, while the least reduction was noted in group F. These differences were not statistically significant (P > 0.05). Conclusion: Within the limitations of this study, Er:YAG laser irradiation combined with fluoride application could not prevent erosion in permanent and primary enamel during the erosive process.