The common method to regulate the bonding strength of hydroxyapatite (HAp) coating and metal substrate by adding low-expansion additives usually sacrifices the biological properties of the coating. To avoid the adverse effects of external additives, HAp coating modified by in situ co-substitution with strontium (Sr) and fluoride (F) ions was prepared on the surface of titanium substrates by electrochemical deposition method. The effects of Sr substitution and F substitution on the coating's morphology, hydrophilicity, electrochemical behavior, in vitro bioactivity, biocompatibility, and bonding strength were investigated. Increased Sr substitution promoted bioactivity, while F substitution induced more regular structure and improved electrochemical behavior. All coating samples were hydrophilic and biocompatible. Sr-F co-substituted HAp coating could improve the bonding strength between the coating and substrates and enhance comprehensive biological properties.