Compare the effectiveness of selected dental lasers for decontamination of machined titanium surfaces in vitro. Seventy-two sterile machined surface titanium discs were individually inoculated with strains of Streptococcus mutans (Sm), Streptococcus oralis (So), Aggregatibacter actinomycetemcomitans (Aa), or all three bacteria together (MIX) at 34.0° C, 20.8% O2 and 5% CO2 for 12 h. After incubation, the discs were divided into six groups: 1) no treatment, 2) 0.12% chlorhexidine gluconate (CHX), and 3) 10,600 CO2, 4) 810 nm diode, 5) 2780 nm Er,Cr:YSGG, 6) 1064 nm Nd:YAG laser groups. After treatment, any remaining viable bacteria were liberated from the discs via sonication, transferred onto brain heart infusion (BHI) agar plates for culturing, and colony-forming units (CFUs) were recorded. Statistical analysis was performed. There were statistically significantly differences (SSD) (p < 0.01) in bacterial reduction of discs individually inoculated with Aa between the Er,Cr:YSGG and Nd:YAG lasers. There was also a SSD (p < 0.01) lower effect with the MIX with the Er,Cr:YSGG compared with all other modalities. Bacterial reduction with the CO2 was better (p < 0.001) than treatment with CHX or the Er,Cr:YSGG laser on killing of So. Although all modalities of treatment showed a mean of 98% or greater viable bacterial reduction, the most consistent bacterial reduction of all titanium discs was with the Nd:YAG laser (100%).