It has been suggested that fluoride products with a protective mechanical component are advantageous in the prevention of erosive wear. The aim of this study was to evaluate in situ the effect of fluoride varnish (FV) in the prevention of wear due to erosion and combined erosion and toothbrush abrasion. Eleven volunteers wore for 3 weeks, during working hours, appliances containing 2 control and 2 FV-treated human enamel samples. Erosion took place extraorally 3 times a day (5 min) in the soft drink Sprite. At the end of each experimental day one control and one FV sample (C-er+abr and FV-er+abr) were brushed (5 s) with fluoridated dentrifice. The remaining control and FV sample (C-er and FV-er) were left unbrushed. Enamel volume loss was quantified by optical profilometry at day 5, 10 and 15. A statistically significant progression in enamel loss was found for the C-er, C-er+abr and FV-er+abr groups (p < 0.001, p < 0.001 and p = 0.001, respectively) but not for the FV-er group (p = 0.053). The values of cumulative normalized volume loss (×108 μm) at day 15 were: C-er 5.53 ± 2.14, C-er+abr 5.70 ± 2.07, FV-er 0.79 ± 0.67 and FV-er+abr 2.76 ± 1.35. The FV-er and FV-er+abr groups showed significant lower volume loss than the C-er group (p < 0.001 and p = 0.005, respectively) and the C-er+abr group (p < 0.001 and p = 0.002, respectively). The results indicate that fluoride varnish is effective in the reduction of erosive wear.