Loguercio AD, Stanislawczuk R, Polli LG, Costa JA, Michel MD, Reis A. . ; 08 2009 Eur J Oral Sci Although it is known that chlorhexidine application may preserve resin–dentin bonds from degradation, the lowest optimal concentration and application time have yet to be established. This study evaluated the effects of different concentrations of chlorhexidine digluconate and different application times on the preservation of resin–dentin bonds formed using two etch-and-rinse adhesives. In experiment 1, after acid etching, the occlusal demineralized dentin was rewetted either with water or with 0.002, 0.02, 0.2, 2, or 4% chlorhexidine for 6065s. In experiment 2, the surfaces were rewetted with water, or with 0.002% or 2% chlorhexidine for 15 or 6065s. After this, both adhesives and composite resin were applied and light-cured. Bonded sticks (0.865mm) were tested under tension (0.565mm65min) immediately or after 665months of storage in water. Two bonded sticks from each tooth were immersed in silver nitrate and analyzed quantitatively using scanning electron microscopy. Reductions in microtensile bond strengths and higher silver nitrate uptake were observed for both adhesives when the rewetting procedure was performed with water. Stable bonds were maintained for up to 665months under all chlorhexidine conditions tested, irrespective of the chlorhexidine concentration and application time. The use of 0.002% chlorhexidine for 1565s seems to be sufficient to preserve resin–dentin interfaces over a 6-month period.