Abstract Background Human gingival fibroblast (HGF-1) cells in the connective tissue provide an effective barrier between the alveolar bone and the oral environment. Cement margins of restorations with intrasulcular preparation or cemented implant restorations are in contact with HGF cells. However, it is unknown to what extend the cement surface finish affects the behavior of HGF cells. The purpose of this study was to compare the behavior of HGF-1 cells in contact with two different resin composite cements with three different surface treatments after light-curing and autopolymerization, respectively. Methods Disks of one adhesive (Multilink Automix, Ivoclar Vivadent [MLA]) and one self-adhesive (RelyX Unicem 2 Automix, 3 M [RUN]) resin composite cement were either light-cured or autopolymerized. Specimen surfaces were prepared with the oxygen inhibition layer intact, polished with P2500-grit silicon carbide paper or treated with a scaler. Cells were cultivated on the specimens for 24 h. Viability assay was performed, and cell morphology was examined with scanning electron microscopy. Additionally, roughness parameters of the specimen were analyzed with a 3D laser scanning microscope. Three-way ANOVA was applied to determine the effect of cement material, curing mode and surface treatment (a = 0.05). Results Overall, cement material (p = 0.031), curing mode (p = 0.001), and surface treatment (p