The effect of a CO2 laser on the structure and permeability of smear layer-covered human dentin was evaluated in vitro. Three different energy levels were used (11, 113, and 566 J/cm2). The lowest exposure to the laser energy increased dentin permeability, measured as a hydraulic conductance, due to partial measured as a hydraulic conductance, due to partial loss of the superficial smear layer and smear plugs. The intermediate energy level also increased dentin permeability by crater formation, making the dentin thinner. The lack of uniform glazing of the surface of the crater, leaving its surface porous and in communication with the underlying dentinal tubules also contributed to the increase in dentin permeability seen with the intermediate laser energy. The highest laser energy produced complete glazing of the crater surfaces and sealed the dentinal tubules beneath the crater. However, it also completely removed the smear layer in a halo zone about 100-microns wide around each crater which increased the permeability of the pericrater dentin at the same time it decreased the permeability of the dentin within the crater. The combined use of scanning electron microscopy and permeability measurements provides important complementary information that is essential in evaluating the effects of lasers on dentin.