A dual-targeting melanin nanoparticle (MNP-TPP-HA) was constructed by grafting the carboxyl groups of hyaluronic acid (HA) and (4-carboxybutyl) triphenylphosphonium bromide (TPP) onto the surface of polyethylene glycol-amino (PEG-NH₂) modified MNP based on 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxy succinimide (NHS) reaction, endowing melanin with the ability of dual active targeting. The fluorescence imaging of in vitro three-dimensional (3D) tumor models and in vivo photoacoustic imaging (PAI) all reveal the excellent target penetration ability of MNP-TPP-HA.