Improved understanding of the effects of hydration and drying in mineralized tissues is highly desirable, particularly for physiologically hydrated biological materials such as dentin. We investigated the influence of hydration on the nanomechanical properties of healthy dentin and hypothesized that drying leads to an increase in indentation induced energy expenditure and hardness. Hydrated and dry dentin were tested with a UMIS set up with a Berkovich indenter at a maximum load of 50 mN. Values representative of the energy expenditure behavior were presented as dissipated energy, U_d, recovered energy, U_e, normalized energy expenditure index, ψ, and hardness, H. Energy expenditure index results, which normalize the energy expenditure for each test and describe the relative energy dissipation–recovery behavior of a material, suggested that, for the relatively severe contact strains about a sharp Berkovich indenter, dissipation dominates the mechanical response of both the hydrated and dry dentin. In support of our initial hypothesis, dry dentin presented a significantly higher energy expenditure index than hydrated dentin (p<0.0001). These results were primarily associated with a lower U_e that was found upon drying. Hydration also decreased H significantly (p<0.0001). In summary, this study presents the first direct measurements of the energy expenditure behavior of hydrated and dry dentin using instrumented nanoindentation.