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Abstract Bacterial colonization plays a key role on the pathogenesis of peri-implantitis and may be influenced by titanium surface topography. The aim of this in vitro study was to evaluate the influence of titanium topography after fluoride ions deposition in the bacterial colonization. Machined (M), double acid-etched (DE) and double acid-etched surface with fluoride ions deposition (Nano F-) were analyzed by scanning electron microscopy, contact angle and roughness (Ra). Streptococcus mutans viability was quantified by Live/Dead Baclight bacterial viability kit. The mean Ra/contact angle values were 0.20 μm/69.13°, 0.53 μm /92.82° and 0.56 μm/94.33° for M, DE and Nano F-, respectively. M surface presented significantly lower live bacterial counts when compared to the Nano F- surface (p=0.007). The dead bacteria count was higher on the Nano F- surface (p=0.001) than on the M and DE surfaces. Crystalline deposition of fluoride ions (Nano F-) promoted an increase in dead bacteria on the tested titanium surface.