A polysaccharide gel containing covalently bound amikacin, a broad-spectrum antibiotic, was produced by using epichlorohydrin-activated hydroxyethyl starch (HES). The structure of the polymers was analyzed by 13C and 1H nuclear magnetic resonance (13C NMR and 1H NMR) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The sites of covalent attachment of amikacin to the epoxypropyl substituent and the HES backbone were determined. The antibacterial activity of the polymer was evaluated in vitro using the agar well diffusion method with the Staphylococcus aureus P209 strain. It was demonstrated that the polymer retained activity in the presence of bacterial amylase, which is released upon bacterial attack. The gel was applied for coating pores and surfaces of a biocomposite material based on a xenogenic bovine bone matrix. In vivo experiments showed the effectiveness of utilizing amikacin-containing biocomposite bone-substitute materials in the treatment of experimental osteomyelitis in rats using objective histological control and X-ray tomography.