Background: Fissure sealants prevent occlusal caries in permanent molars. Enamel preparation methods are used before fissure sealing. Aims: To investigate effects of bioglass air-abrasion pre-treatment with and without an adhesive, on fissure enamel of permanent teeth, with respect to etchability, microleakage and microtensile bond strength. Design: Half of the occlusal surfaces of 50 extracted premolars underwent bioglass air-abrasion. Dye was applied to the entire occlusal surface. Photographs were taken to score etched surface by dye uptake. Adhesive was applied to 25 of the bioglass-treated areas and all teeth were fissure sealed, sectioned, and evaluated using confocal microscopy. Buccal and lingual surfaces of a further eight premolars were acid-etched and randomly received: air-abrasion, adhesive, both, or none before sealant application for microtensile bond strength measurement in half of the samples immediately and half following 6 months of water immersion. Results: Linear mixed models and multinomial logistic regression were used (P = 0.05). Bioglass air-abrasion significantly improved enamel etchability and reduced microleakage. The addition of an adhesive made no difference to either microleakage or microtensile bond strength. The combination of bioglass abrasion and adhesive led to more cohesive, rather than adhesive, failure. Conclusions: Bioglass air-abrasion improved enamel etchability and reduced microleakage irrespective of the adhesive use but neither pre-treatment affected the microtensile bond strength.