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OBJECTIVES This in situ study aimed to analyze the influence of different resin cement removal techniques on bacterial adhesion and biodegradation at the marginal interface of ceramic laminates. METHODS AND MATERIALS Eighty feldspathic ceramic (F) blocks were prepared and cemented onto bovine enamel slabs (7×2.5×2 mm). Excess cement was removed using a microbrush (MBR), a scalpel blade (SCP), or a Teflon spatula (TSP). For the biodegradation analysis, 40 disc-shaped resin cement specimens were prepared (7×1.5 mm) using a Teflon mold. The specimens were randomly allocated into two groups: (1) No finishing procedure (only Mylar strip), and (2) with finishing and polishing procedures using the Jiffy system (Ultradent, South Jordan, UT, USA) (n=20). The in situ phase consisted of using an intraoral palatal device by 20 volunteers for 7 days. Each device contained five cylindrical wells (8×3 mm), where three dental blocks and two cement specimens were included in the wells. Surface roughness (Ra) was measured using a contact profilometer. A micromorphological analysis was performed under a stereomicroscope and a scanning electron microscope. Bacterial adhesion was quantitated based on the number of colony-forming units (CFU/mL) and their biofilm development potential. RESULTS The cement removal techniques directly affected surface roughness at the marginal interface (p<0.001), and the SCP technique produced higher mean roughness, regardless of the surface area analyzed. Surface polishing protected cement specimens from further biodegradation (p=0.148). There were no differences in CFU counts between the groups after the in situ phase (p=0.96). All specimens showed CFU with a strong ability to develop a biofilm. CONCLUSIONS The techniques used for cement removal increased the surface roughness of ceramic laminates, particularly SCP, but they did not affect bacterial adhesion at the marginal interface. Surface polishing of the resin cement is recommended to mitigate biodegradation.