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Aim: To evaluate in vitro compressive strength of fluoride-containing resin-based sealant (F) before and after fluoride release in water. Materials and Methods: We used 40 specimens with 6 mm of height and 20 mm of diameter using fluoride-containing resin-based sealant (FluroShield,Brasil, Dentsply) and non-fluoride-containing resin-based sealant (Concise (TM) Light Cured White Sealant, USA, 3M ESPE). The specimens were divided into four groups, two fluoride-containing resin-based sealants (F1 and F2) and two like control groups (C1 and C2). A group of specimens of fluoride-containing resin-based sealant and a control group (F1/C1) remained without exposure to water, while other groups (F2/C2) were immersed in distilled water for 30 days. The release of fluoride from the F2 was measured through selective electrode method on days 1, 2, 3 and 30. Subsequently, the compressive strength was measured using a universal testing machine (Lloyd, LR 100, UK) with a crosshead speed of 1 mm/min. Comparisons between groups F1 vs F2 and C1 vs C2 were analyzed with t-Student. The significance level was set at 0.05. Results: The compressive strength (Mpa) before and after immersion in water for fluoride-containing resin-based sealant was 337.2 and 337.4, while the control sealant was 203.8 and 213.4. For both groups the observed differences were not significant. The initial fluoride release was 1.9 ppm during the first 24 hours and then declined to 0.0 ppm during the third day of release. Conclusion: In this in vitro study, the release of fluoride from a fluoride-containing resin-based sealant does not affect compressive strength.