OBJECTIVES: To investigate if differences in titanium implant surface topography influence biofilm formation. MATERIALS AND METHODS: Titanium discs were prepared and characterized using a profilometer: Group A ( 0.15 µm, smooth), Group B ( 0.64 µm, minimally rough) and Group C ( 1.3 µm, moderately rough). Contact angle and surface free energy (SFE) were determined for each group. Non-preconditioned titanium discs were incubated with for 24 h. In additional experiments, the titanium discs were initially coated with human saliva, bovine serum albumin or phosphate-buffered saline for 2 h before incubation with for 24 h. The amount of fungal biofilm formation was quantified using a colorimetric assay. RESULTS: biofilm formation was significantly lower ( < 0.05) on the minimally rough titanium surface compared to smooth and moderately rough surfaces. The titanium surface displaying the lowest SFE (Group B) was associated with significantly lower ( < 0.05) biofilm formation than the other two groups. Salivary coating resulted in greater adherence of with increased surface roughness. CONCLUSIONS: The minimally rough titanium discs displayed lowest SFE compared to smooth and moderately rough surfaces and showed the least biofilm formation. This study demonstrated that biofilm formation increased in a SFE-dependent manner. These findings suggest that SFE might be a more explanatory factor for biofilm formation on titanium surfaces than roughness. The presence of a pellicle coating may negate the impact of SFE on biofilm formation on titanium surfaces.