Purpose: The purpose of the present study was to compare the Vickers hardness numbers (VHNs), roughness numbers (RNs) and biological compatibility of glass carbomer (Glass Fill), resin-modified glass ionomer (Fuji II LC) and self-adhering flowable composite (Vertise Flow) materials. Materials and methods: Disc-shaped specimens of test materials (n = 15/group) were prepared, and VHNs and RNs were determined after 24 h. A direct contact test was used for cytotoxicity evaluation. Cell viability was measured for 24 h post-exposition with a photometric test (MTT assay; n = 16). Data were analysed using one-way analysis of variance with a post hoc Tukeys test, the Kruskal-Wallis test and the Mann-Whitney U-test (p < 0.05). Results: Fuji II LC had the highest VHN. The VHN of Fuji II LC differed significantly from those of the other materials (p < 0.05). The RNs of Fuji II and Glass Fill were higher than that of Vertise Flow (p < 0.05). The self-adhering materials were not significantly cytotoxic compared with the control group (p > 0.05). Conclusions: The materials tested in this study showed a similar lack of cytotoxicity. The VHN of Fuji II LC was the highest, and the RN of Vertise Flow was the lowest.