Background: The long-term capability of resin sealants and glass ionomer cements to release fluoride is associated to a reduction in pit and fissure caries. The regular use of fluoride varnishes/toothpastes can result in the absorption of fluoride into the sealant. The objective of the present study was to assess the fluoride release/uptake capacities of different fissure sealants. Material and Methods: Three different fissure sealants (Fuji Triage/GC, Fissurit FX/Voco and Grandio Seal/Voco) were examined. Ten discs of each material were prepared. Each disc was incubated with distilled water and then the solution analyzed for diluted for fluoride concentration, using a combination of fluoride electrode (OrionGP 1 S/N 13824, Orion Research Inc, Boston, MA, USA) connected to an expandable ion analyzer (Orion 720A, Orion Research Inc, Boston, MA, USA). Standard curves between 1 and 100 ppm F- were used to calibrate the electrode. Cumulative fluoride release was measured on days 1, 2, 3, 5, 7, 21, 35 and 49, then two different fluoride varnishes/ pastes (Profluorid Varnish/Voco, MI Paste Plus/GC), were applied to the sealants tested, and fluoride release (after reuptake) was measured on days 56, 70 and 84. Results: Kruskal Wallis test confirmed significant differences in fluoride release between Fuji Triage/GC and Fissurit FX/Voco and Grandio Seal/Voco from day 1 ( P < 0.001). The application of fluoride varnish Profluorid Varnish enhanced the fluoride release for all sealants ( P < 0.05). MI Paste Plus enhanced the fluoride release for all sealants except for Fuji Triage/GC ( P > 0.05). Conclusions: The GIC-based sealant (Fuji Triage/GC) released significantly more fluoride than the resin sealants tested. The exposure to the fluoridated varnish (Profluorid Varnish) significantly recharged the sealants tested more than the CPP-ACPF toothpaste (MI Paste Plus).