Most non-traumatic fractures occur in teeth that have been treated, for example restored or endodontically repaired. It is therefore essential to evaluate the structure and mechanical properties of altered forms of dentin. One such altered dentin is transparent (sclerotic) dentin, which forms gradually with aging. Accordingly, in the present study, we seek to study differences in the structure, i.e., dentinal mineral concentration, mineral crystallite size, and the mechanical properties, i.e., elastic moduli, fracture toughness and fatigue behavior, of normal and transparent root dentin. The mineral concentration, measured by x-ray computed tomography, was found to be significantly higher in transparent dentin, with the majority of the increase being due to the closure of the tubule lumens. Crystallite size, as measured by small angle x-ray scattering, appeared to be slightly reduced in transparent dentin, although the difference was not statistically significant. The elastic properties remained unchanged, although transparent dentin showed almost no yield/post-yield behavior. The fracture toughness was lowered by roughly 20%, while the fatigue resistance was deleteriously affected at high stress levels. These results are discussed in terms of the altered microstructure of transparent dentin.