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This paper describes a promising electrophoretic deposition (EPD) approach for developing composite coatings based on chitosan with the additional components of hydroxyapatite, graphene, and gentamicin on titanium substrate. Bioactive properties were investigated in vitro by immersing the coatings in simulated body fluid (SBF) at 37 °C. The newly formed biomimetic layer on the top of the deposited chitosan-based coatings on Ti was confirmed by X-ray diffraction, Fourier transform infrared spectroscopy, field emission scanning electron microscopy and electrochemical measurements, while coatings' bioactivity was proved by alkaline phosphatase activity assay in MRC-5 and L929 tested cell lines. The biocompatibility towards MRC-5 and L929 cell lines was investigated by dye exclusion test (DET) implying the non-cytotoxic effect of coatings. Gentamicin release studies were monitored during 21-day by high-performance liquid chromatography coupled with mass spectrometry, indicating rapid release of gentamicin (≈40%) in the first 48 h and more than 60% after 14 days.