This study determined the effect of thiourethane-functionalized fillers (TU) on the antimicrobial properties, cytotoxicity, degree of conversion (DC), water sorption (Wsp) and solubility (Wsl) of experimental composites. TU-modified fillers were added at different ratios in experimental composites: 0 (Control-TU0), 25% (TU25), 50% (TU50), 75% (TU75) and 100wt% (TU100). The antimicrobial properties were detected through the exhaustion test and counting of Streptococus mutans colonies for biofilm formation. Cytotoxicity to human gingival fibroblasts was evaluated in three different parameters: XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide), NRU (Neutral Red Uptake assay) and CVDE (Crystal Violet Dye Exclusion test)) at the same cells. ELISA was used to measure the IL-6 and b-FGF biomarkers. DC was determined by Fourier-transformed infrared spectroscopy, while Wsp and Wsl by mass variations. Inhibitory capacity of biofilm formation was not observed for any material. All groups presented at least 70% of cell survival within the observed periods (24h and 7 days). Positive control (toxic) had high IL-6 values and low b-FGF values. No significant variations in DC, Wsp, and Wsl were observed among the experimental groups. The use of thiourethane did not present antimicrobial and cytotoxic activity and the tested materials presented equivalent properties to those conventionally used in dentistry.