Thesis submitted in fulfilment of the requirements for the degree Master of Technology: Biomedical Technology In the Faculty of Health and Wellness Sciences At Cape Peninsula University of Technology 2013 / Newly developed and commercial dental resins which are commonly used nowadays have to be tested for their antimicrobial susceptibility. The purpose of this in vitro study was to investigate the antimicrobial activity of a titanium oxide (TiO2) nano-composite which was prepared with different antibacterial substances and used as restoratives in dentistry to combat certain selected bacteria that are considered the principle causes of some tooth diseases, for example, tooth decay and to prevent unsuccessful dental restoration. The TiO2 nano-composite was prepared and divided into four groups: The first group was an untreated TiO2 nano-composite. The second group was silane-treated TiO2 nano-composite. The third group was treated TiO2 nano-composite which was combined with chlorhexidine gluconate (CHxG). The fourth group was treated TiO2 nano-composite which was combined with benzalkonium chloride (BzCl). Five of the selected bacteria were grown overnight in Petri dishes. Four of them, namely, Escherichia coli (E. coli) ATCC 11775, Staphylococcus aureus (S. aureus) ATCC 12600, Enterococcus faecalis (E. faecalis) ATCC 29212, and Pseudomonas aeruginosa (P. aeruginosa) ATCC 10145, were grown on Müller-Hinton Agar (MHA). Streptococcus mutans (S. mutans) ATCC 25175 was grown on Brain Heart Infusion (BHI) agar. All these bacteria were tested against the TiO2 nano-composite, and incubated for 24 hours at 37°C, except S. mutans, which was incubated separately and exposed to CO2. It was placed into a CO2 water-jacketed incubator in an atmosphere of 5% CO2 for 24 hours at 37°C. The obtained results showed that neither of the groups of TiO2 nano-composites, (untreated TiO2 nano-composite and treated TiO2 nano-composite) exhibited antimicrobial activity against the pathogens. Only preparations of TiO2 nano-composites at a concentration of 3 %m/m of both CHxG and BzCl showed antimicrobial activity against S. aureus. Antimicrobial activity against S. mutans, E. coli, P. aeruginosa, E. faecalis and S. aureus, were only realized at a concentration of 10 %m/m for both CHxG and BzCl..