Dentine hypersensitivity still represent a major clinical problem only partially solved. The presence of exposed opened dentinal tubules has been demonstrated increase the dentine permeability and it is the responsible for pain and sensitivity. Stannous fluoride solution has been recently proposed to reduce dentine hypersensitivity. The aim of the present study was to evaluate the dentinal permeability of a new stannous fluoride gel proposed for the therapy of dentine hypersensitivity. Human extracted teeth have been used and mounted and connected with a hydraulic pressure apparatus working at 1 psi (70 cm of water pressure). It was calculated the permeability of untreated smear layer, treated smear layer (after gel application) and after acidic treatment. It was observed that gel treatment was able to reduce dentine permeability. Scanning electron microscopy demonstrated the presence of a homogeneous smear layer after treatment with gel. This in vitro study confirms that stannous fluoride treatment has the capacity to modify dentine permeability of sensitive dentine.