Hydroxyapatite (HA) is a calcium phosphate-based inorganic compound that is similar to the mineral composition of bones and teeth. The large surface area due to the nano-sized HA and its structure containing positive and negative charges on the sides can facilitate absorption for biomolecules. HA has good characteristics such as biocompatibility, stability, and bioactivity properties, which leads to wide development for internal support materials in electrochemical biosensors construction. The HA in the electrochemical biosensor also can be used as a signal probe besides as an immobilization matrix. This review describes the several applications of hydroxyapatite in nucleic acids and aptamers, antibodies, and enzyme-based biosensors followed by electrochemical methods measured in the last decade.