The purpose of this in vitro study was to evaluate the potential remineralization of enamel and dentine erosion lesions after the application of five different toothpastes. A total of 104 enamel and dentine samples were prepared from maxillary third molars. Each group was divided according to the toothpaste application mode (topical = 56; brushing = 48) and the toothpaste used seven topical groups and six brushing groups (n = 8). The groups included negative control (NC), positive control (PC), Sensodyne Pronamel (SP), Regenerate (R), Regenerate with boosting serum (R+), Colgate Duraphat 5000 (CD), and tooth mousse (TM). The statistical analysis showed significant surface microhardness (SMH) change. All enamel groups showed a significant decrease in SMH compared to NCfor both application modes. However, no significance was recorded between test groups. Similar results were observed between dentine groups and their relevant controls for both application modes, except brushed Rand R+groups, which were insignificant to their NC. For topical groups, TMshowed a significant increase in SMH. While Rand R+showed lower loss than SPand CD. All tested agents offered a degree of remineralization in both enamel and dentine with no significant difference between agents in enamel groups while R, R+, and TMoffered better results in dentine groups. For dentine groups, similar findings were observed with superior tooth surface protection with the application of TM over other agents. Tooth surface remineralization was achieved when agents were either applied topically or brushed over the surface.