In this study, TiO2 films were deposited via the doctor blade technique on fiber-cement surfaces. Two types of nanoparticles (TiO2-P25 from Degussa and TiO2-PC105 from Tronox) were used to produce films. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) images revealed films with homogeneous and nanoparticulated morphology. The TiO2 PC105 film presented a lower roughness parameter (RMS) in relation to that of the TiO2 P25-based film. Both films exhibited high hydrophilicity when exposed to UV-A radiation (contact angle θ < 6°). The photocatalytic activity of the films was evaluated by standardized methylene blue dye degradation assays under UV-A irradiation (1.0 mW/cm2). The TiO2-PC105 film showed a photonic efficiency of ξ = 0.1%, while for the films obtained with TiO2-P25, ξ = 0.08%. The cement surface modified with the PC105 film was evaluated for antimicrobial activity through the use of multiple pathogens commonly found in hospitals. A considerably high efficiency was measured with visible light. Growth inhibition rates of 99.0% ± 0.2, 99.1% ± 0.2, 99.1% ± 0.2, 97.5% ± 0.5, 98.0% ± 0.5 and 98.0% ± 0.5 were found for Staphylococcus aureus, Klebsiella sp., Escherichia coli, Rhizobium sp., Fusarium sp. and Penicillium sp., respectively. The results show the self-cleaning ability and their potential use for protection, by preventing contamination of the fiber-cement surface and opening new possibilities for the use of this building material.