To ensure clinical success, the implant and the surrounding bone tissue must not only be integrated, but also must not be suspected of infection. In this work, an antibacterial and bioactive nanostructured calcium silicate (CaSi) layer on titanium substrate by an electrospray deposition method was prepared, followed by annealing at 700, 750 and 800 °C to improve the bonding strength of the CaSi coating. The phase composition, microstructure and bonding strength of the CaSi coatings were examined. Human mesenchymal stem cells (hMSCs), Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus) species were used to analyze the osteogenic and antibacterial activity of the coatings, respectively. Experimental results showed that the as-prepared CaSi coating was mainly composted of β-dicalcium silicate phase with a particle size of about 300 nm. After annealing, the thickness of the oxidation reaction layer increased obviously from 0.3 μm to 1 μm with increase in temperature, which was confirmed by the cross-sectional morphology and element depth profile. The bonding strength of the coating annealed at 750 °C (19.0 MPa) was significantly higher (p < 0.05) than that of the as-prepared coating (4.4 MPa) and the ISO 13,779 standard (15 MPa). The results of antibacterial efficacy and stem cell osteogenesis consistently elaborated that the 750 °C-annealed coating had higher activity than the as-prepared coating and the Ti control. It is concluded that after annealing at 750 °C, the CaSi nanoparticle-coated Ti implant had good bond strength, osteogenic and antibacterial activity.Copyright © 2021 Elsevier B.V. All rights reserved.