Bonding interface and dentin enzymatic activity of two universal adhesives applied following different etching approaches

Objectives: Etching approaches [37% phosphoric acid, self-etching, 10–3 solution (3% ferric chloride dissolved in 10% citric acid), or 1.4% nitric acid] were evaluated regarding enamel shear bond strength (24 h), dentin microtensile bond strength (24 h and 2 years), failure mode, enzymatic activity of the hybrid layer, and nanoleakage (24 h and 2 years) of Prime&Bond Universal (PBU, Dentsply-Sirona) and Gluma Bond Universal (GBU, Kulzer). Methods: Adhesives were applied on blot-dried (wet-bonding, positive control) or air-dried (remaining groups) dentin after acid-etching (15 s) or in self-etch mode. Enamel and dentin bond strength tests used 160 human teeth (n = 10). Failure mode of tested samples and nanoleakage within the dentin-adhesive interface (n = 5) were analyzed by scanning electron microscopy. Dentin enzymatic activity was investigated by in situ zymography (n = 3). Results: Enamel bond strengths did not differ statistically among groups. Wet-bonding with 37% phosphoric acid showed similar dentin bond strength compared to 10–3 solution or 1.4% nitric acid at 24 h for both adhesives. None of the etchants inhibited enzymatic activity, and all groups showed dentin bond strength reduction after 2-year storage. GBU showed higher nanoleakage. Experimental etchants did not affect enamel bond strength. Dentin bond strength was not stable after 2 years, despite promising 24-hour results. Significance: This study suggests multiple etching approaches to optimize and achieve stable dentin bonding, while also offering in-depth information about the performance of recently released universal adhesive systems.