In this chapter, the current progress on nanoindentation measurement of dental hard tissues is summarized. With the help of nanoindentation techniques, it is possible to measure the mechanical properties of small volumes of dental hard tissues at the micro- and nano-level with respect to their unique microstructures, namely the rod unit of the enamel and tubule structure of dentin. The results indicate that the microstructure and organic components regulate the mechanical behaviour of these natural biocomposites to a great extent. Furthermore, application of nanoindentation to the investigation of diseased or defective states of tooth tissues, dental treatments and dental materials is also introduced and discussed.