Aim: To investigate in situ/ex vivo the effect of remineralizing agents in the prevention of dental erosion in permanent and primary teeth. Study design: A randomized, controlled, double-blind study with crossover design with three treatment phases: Control, ProNamel (R) and Tooth Mousse (TM). Twenty adults and children wore removable palatal appliances containing two insets of permanent and primary human enamel and used the corresponding assigned toothpaste twice daily for 10 days. The enamel samples were then removed, mounted on acrylic bases and acid-challenged in demineralizing solution. Enamel surface microhardness (S mu H) was measured pre and post acid challenge. Data were analyzed using two-way ANOVA and Tukey's post hoc test (P < 0.05). Results: The mean S mu H values (Vicker's unit) prior to acid challenge were: Permanent teeth (Control 366.16 perpendicular to 12.28, ProNamel (R) 372.18 perpendicular to 14.75, Tooth Mousse (TM) 370.19 perpendicular to 11.88) and Primary teeth (Control 325.31 perpendicular to 11.90, ProNamel (R) 327.34 perpendicular to 9.90, Tooth Mousse (TM) 331.63 perpendicular to 10.55). Following the acid challenge, the mean %.Delta S mu H (perpendicular to SD) were: Permanent (79.72 perpendicular to 1.59, 66.52 perpendicular to 2.45, 60.13 perpendicular to 4.98) and Primary (81.09 perpendicular to 2.90, 76.50 perpendicular to 3.13, 69.02 perpendicular to 4.23). Conclusion: The application of remineralizing agents reduced the significantly softening by acidic attack of enamel especially in the permanent dentition.