Objectives This study investigated biofilm formation on discs of metal alloys, zirconia and polyetherketoneketone in vivo. Material and Methods Sixteen healthy volunteers conducted two runs of 24 hr each wearing an intraoral splint with 15 discs representing five different materials (gold‐based [EL] and silver‐based [PA] noble metal alloys; zirconia [ZR]; polyetherketoneketone [PEKK]; titanium zirconium alloy [TiZr]). Safranin staining assays and colony‐forming unit (CFU) counts were conducted. Linear mixed‐effects models were used to compare materials, and geometric mean ratios with 95% confidence interval were calculated with the level of significance set at α = 0.05. Results Less biofilm mass and lower CFU counts were found on PA and EL, while ZR and PEKK developed similar levels as the reference material TiZr alloy. Compared with PA, biofilm mass was 1.5 times higher for EL (p = .004), 1.7 times higher for PEKK (p < .001), 2.2 times higher for TiZr (p < .001) and 2.4 times higher for ZR (p < .001). The culturing method confirmed these results for EL and PA with lower CFU compared to TiZr. The biomass staining technique and cell culturing correlated for EL and PA. Conclusion Silver‐based noble alloy and gold‐based high noble alloy demonstrated the least biofilm formation indicating a potential clinical use as material for implant components in the transmucosal compartment. Zirconia and Polyetherketoneketone revealed similar results as the reference material titanium zirconium alloy used in commercially available titanium dental implant.