The development of nanomaterials based antibacterial coatings has received considerable attention towards the control and prevention of healthcare associated infections (HAIs). In this article, we are reporting the preparation of titanium oxide (TiO2) nanopaint by embedding the TiO2 nanoparticles in alkyd resin matrix (with other suitable non-toxic additives) using ball milling process. The Fourier transform-infra red spectroscopy and scanning electron microscope analysis revealed the incorporation of TiO2 nanoparticles in the alkyd resin matrix of the paint. The TiO2 nanopaint coatings are examined for their antibacterial properties against different types of bacterial strains viz. Pseudomonas aeruginosa, Staphylococcus aureus, and methicillin resistant Staphylococcus aureus (MRSA). The experimental results showed that the TiO2 nanopaint coated surfaces inhibited about 85%, 90% and 76% of microbial population against Pseudomonas aeruginosa, Staphylococcus aureus and MRSA, respectively. We believe that our efforts ensure the potential applications of the TiO2 nanopaint in the prevention of HAIs.