This in vitro study evaluated the effects of Nd:YAG and Er:YAG lasers on reducing dentin permeability by sealing opened tubules. According to hydrodynamic theory, dentine hypersensitivity occurs when dentin is exposed with tubules opened. Consequently, a painful sensation occurs due to an intensification of the dentinal permeability. Treatment, therefore, should be based on a decrease of this permeability, achieved by the obliteration of dentinal tubules. The Nd:YAG laser is known for its capacity to seal dentinal tubules; however, few studies concerning treatment with Er:YAG laser are available. The Nd:YAG laser was used, based on two parameters: (A) 1.0 W, 10 Hz, and (B) 1.5 W, 15 Hz. The Er:YAG laser was used at 60 mJ, 2 Hz, four applications of 20 sec each, at 6 mm from the surface. After irradiation, all samples were immersed in 1% Rodamine B dye solution, in order to evaluate the penetration of the dye solution and observe the decrease/increase of dentinal permeability after the laser treatment. The laser conditions used in the present study decreased the permeability as follows: (a) when using the Er:YAG laser at 60 mJ, 2 Hz in 26.05%, and (b) when using the Nd:YAG laser at 1.5 W, 15 Hz in 19.03%--with no statistical difference between them. Additionally, the Nd:YAG laser at 1.0 W, 10 Hz, decreased permeability in 4.59%, with a smaller effect. According to the results of this in vitro study, the Er:YAG laser at 60 mJ, 2 Hz, and the Nd:YAG laser at 1.5 W, 15 Hz are useful for decreasing dentin permeability.