Background <p>The clinical longevity of pit and fissure sealants is closely related to their physical and chemical properties. To address the moisture sensitivity of hydrophobic pit and fissure sealants, hydrophilic types were introduced; however, their performance is not yet fully understood.</p> Objectives <p>The present study was carried out to compare and evaluate microleakage, penetration depth, and fluoride release of three different pit and fissure sealants.</p> Methods <p>Thirty extracted permanent upper first premolars were randomly distributed to three groups: Embrace WetBond, Denuseal, and Helioseal F Plus, each with or without bonding application (<i>n</i> = 5). Microleakage and penetration depth were evaluated using dye penetration, stereomicroscope examination, and image analysis software. Fluoride release was measured over four weeks using the SPANDS colorimetric method. SPSS version 27.0 was used for statistical analysis, and significance was set at <i>P</i> &lt; 0.05.</p> Results <p>Bonding application did not significantly reduce microleakage or improve penetration depth for any of the tested materials (<i>p</i> &gt; 0.05). Helioseal F Plus demonstrated significantly lower microleakage and greater penetration under both bonded and non-bonded conditions compared with Denuseal (<i>p</i> &lt; 0.05). Embrace WetBond showed higher microleakage and lower penetration depth than Helioseal F Plus. All tested pit and fissure sealants exhibited a pronounced initial fluoride release followed by a significant decline over time (<i>p</i> &lt; 0.001). Embrace WetBond consistently gave the highest fluoride values among the three materials from Week 1 onward (<i>p</i> &lt; 0.05).</p> Conclusion <p>Helioseal F Plus demonstrated superior sealing ability with minimal microleakage and satisfactory penetration. Embrace WetBond exhibited enhanced fluoride release patterns.</p>