Objective To investigate the variations of elastic modulus and hardness of human dentin with aging and location. Methods The non-carious third molars were collected and divided into youth group, middle-aged group and elderly group. Nanoidentation test was conducted on the dentin specimens from multiple regions to test their mechanical properties. Results The elastic modulus and hardness in outer and middle dentin were greater than those in inner dentin; the elastic modulus and hardness increased with the aging in each region of the dentin. Conclusions The dentin shows a gradient mechanical property. The stiffness and hardness in middle and outer dentin are greater than those in inner dentin. The deformation resistance in middle and outer dentin is stronger than that in inner dentin. Meanwhile, elastic modulus and hardness of the dentin increase with the aging.