PURPOSE: To evaluate whether patients should be advised to perform toothbrushing before or after an acidic challenge to minimize enamel and dentin wear by brushing abrasion. METHODS: The study was a two-period crossover design (A and B, each 14 days) in which three enamel and dentin specimens were fixed in intraoral appliances of 10 volunteers. The following regimens were performed three times a day with at least 4 hours in between: A: 20-second brushing treatment in an automatic brushing machine, 5 minutes intraoral exposure of the specimens, extraoral erosion of enamel and dentin specimens for 40 seconds or B: Extraoral erosion for 40 seconds, 5 minutes intraoral exposure of the specimens, 20-second brushing treatment in an automatic brushing machine. Enamel and dentin loss at the end of each 14-day regimen was assessed by profilometry and statistically analyzed by t-test. RESULTS: For all volunteers, mean enamel and dentin wear was significantly lower when brushing treatment was performed before erosion (A: enamel: 2.3 +/- 1.0 microm, dentin: 4.1 +/- 1.6 microm) than when brushing was applied after erosion (B: enamel: 6.4 +/- 3.0 microm, dentin: 15.3 +/- 6.8 microm). It was concluded that patients awaiting an erosive attack should perform toothbrushing prior to rather than after an acidic challenge to minimize enamel and dentin wear.