Surface topography and physical-chemical properties usually play a key-role in both osseointegration improvement and bacterial colonization reduction over the surface of dental implants. The aim of this study is to compare the chemical and bacteriological behavior of two different acid passivation surface treatments on titanium c.p. grade 3 used for dental implant manufacturing. Surface roughness was evaluated using White Light Interferometry (WLI) in order to determine different roughness parameters such as average roughness (Sa), the spacing parameter (Sm) and the hybrid parameter of surface index area (SIA). Contact angle (CA) and surface free energy (SFE) were evaluated in order to establish the surface wettability of the different groups of samples. Titanium ion-release from the different samples was also been analyzed in Hank’s solution medium at 37 °C by using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) at different immersion times. Bacterial viability adhesion assays were done using S. sanguinis (CECT 480, Spain) as a bacterial strain model of primary colonizer in oral biofilm formation. The bacteria attachment and morphology on Ti surfaces were determined using a live/dead staining method after 4 h of incubation and further analyzed by scanning electron microscope (SEM). Acid passivation surface treatments produced a statistically-significant (p < 0.05) roughness increase in all the evaluated parameters (Sa, Sm, SAI). The treatment with citric acid decreased the static contact angle (CA) and caused an increase in surface free energy (SFE) with a high polarization and oxidizing character. These physical-chemical surface characteristics obtained by means of citric acid passivation caused the bactericidal behavior as it has been proved in bacterial studies.