BackgroundGreater degradation of the hybrid layer is expected when a universal adhesive system is used, especially in the conventional application strategy. Therefore, it would important to evaluate the effect of the ethanol (ETH) and a potential matrix protease inhibitor (caffeic acid phenethyl ester/ CAPE) to maximize the ability to achieve stable dentin bond strength. The aim of this study was to evaluated the effect of ETH on a wet-bonding technique, and dentin pretreatments with different concentrations of CAPE in ethanolic solution, followed by application of a universal adhesive system (Single Bond Universal) to inhibit proteolytic activity.Material and MethodsDentin blocks were allocated to eight experimental groups according to the strategy (total-etch our self-etch) and treatments: ETH, or dentin pretreatment with CAPE (at 0.5%, 2.5%; and 5.0%). Half of each block (each hemiblock) served as the control (without dentin pretreatments) for the same group. The bonding strategy was performed (adhesive system/ restoration with composite resin). Two slices were obtained from each hemiblock and evaluated using in situ zymography. The proteolytic activity was analyzed by quantifying the green photons of the images obtained under a fluorescence microscope in three dentin locations close to the dentin-resin interface: hybrid layer (HL), underlying dentin (UD) and deep dentin (DD).ResultsWilcoxon tests (for comparison between experimental and control groups) and Friedman and Nemenyi tests (for comparisons between interface locations) showed that there was no difference between the groups with different CAPE concentrations and the respective control groups (p>0.05). ETH reduced the proteolytic activity at the HL and UD (p<0.05).ConclusionsThe wet-bonding technique with ETH proved effective in reducing the proteolytic activity. The use of CAPE in different concentrations solubilized in ethanol did not have a favorable effect on proteolytic inhibition.Key words:Adhesives, Hybrid layer, Dentin, Metalloproteinases.