Titanium alloys are widely used for biomedical implants, but their performance is limited by wear, corrosion, and susceptibility to bacterial colonisation. To overcome these drawbacks, multilayer Ti–Cu oxide coatings were deposited on Ti6Al4V substrates using direct current magnetron sputtering. Two multilayer architectures (6 × 2 and 12 × 2 TiO₂/CuO bilayers) were fabricated and evaluated for their structural, mechanical, electrochemical, and biological properties. SEM/EDS and XRD confirmed well-adhered crystalline coatings consisting of rutile/anatase TiO₂ and monoclinic CuO with uniform elemental distribution. The coatings increased surface roughness, improved adhesion, and enhanced hardness by up to ~180% compared to uncoated Ti6Al4V alloy. Compared to the bare substrate, electrochemical testing in simulated body fluid showed higher corrosion resistance of both coated samples, but particularly for the 12 × 2 multilayers. Both architectures provided sustained Cu²⁺ release over seven days without a burst effect. In vitro biological testing showed that both multilayer coatings achieved over 96% inhibition of Gram-positive bacteria such as Staphylococcus aureus and Bacillus subtilis, while exhibiting moderate antibacterial effects against Gram-negative strains (Escherichia coli, Pseudomonas aeruginosa). Despite the presence of copper, MG-63 osteoblast-like cells demonstrated sustained viability and successful extracellular matrix mineralisation, indicating excellent cytocompatibility of the coatings with bone-forming cells. These results demonstrate that multilayer Ti–Cu oxide coatings can effectively balance antibacterial performance, corrosion resistance, mechanical strength, and support bone cell integration, making them a promising strategy for the surface modification of titanium-based biomedical implants.