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The aim of this in vitro study was to investigate the effect of hydrogen peroxide (H₂O₂) on the surface properties of various zirconia-based dental implant materials and the response of human alveolar bone osteoblasts. For this purpose, discs of two zirconia-based materials with smooth and roughened surfaces were immersed in 20% H₂O₂ for two hours. Scanning electron and atomic force microscopy showed no topographic changes after H₂O₂-treatment. Contact angle measurements (1), X-ray photoelectron spectroscopy (2) and X-ray diffraction (3) indicated that H₂O₂-treated surfaces (1) increased in hydrophilicity (<i>p</i> < 0.05) and (2) on three surfaces the carbon content decreased (33–60%), while (3) the monoclinic phase increased on all surfaces. Immunofluorescence analysis of the cell area and DNA-quantification and alkaline phosphatase activity revealed no effect of H₂O₂-treatment on cell behavior. Proliferation activity was significantly higher on three of the four untreated surfaces, especially on the smooth surfaces (<i>p</i> < 0.05). Within the limitations of this study, it can be concluded that exposure of zirconia surfaces to 20% H₂O₂ for 2 h increases the wettability of the surfaces, but also seems to increase the monoclinic phase, especially on roughened surfaces, which can be considered detrimental to material stability. Moreover, the H₂O₂-treatment has no influence on osteoblast behavior.