The AFM combined nanoindentation was performed to observe the ultrastructure of enamel rod from various section plans and positions while probing their mechanical and tribological properties of the area. The nanohardness and the elastic modulus of the head region of the enamel rods are significantly higher than that of the tail region and the axial-sectional plane. Both nanohardness and elastic modulus gradually decrease from enamel surface toward dentino-enamel junction. Such a variation correlates well with the decreasing trend of calcium composition from our element analysis. The friction coefficient and nanowear of the enamel showed an inversed trend to the hardness with respect to their relative topological position in the long axis of enamel rod toward DEJ. The relationship between the nanowear depth and the distance from the outer enamel surface to DEJ presented exponential function. The results presented clarify the basic nanomechanical and nanotribological properties of human enamel rods and provide a useful reference for the future development of dental restorative materials.

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