This in vitro study evaluated the effect of myristyltrimethylammonium bromide (MYTAB) on the physical, chemical, and biological properties of an experimental dental resin. The resin was formulated with dental dimetacrylate monomers and a photoinitiator/co-initiator system. MYTAB was added at 0.5 (G 0.5% ), 1 (G 1% ), and 2 (G 2% ) wt %, and one group remained without MYTAB and was used as the control (G Ctrl ). The resins were analyzed for the polymerization kinetics, degree of conversion, ultimate tensile strength (UTS), antibacterial activity against Streptococcus mutans, and cytotoxicity against human keratinocytes. Changes in the polymerization kinetics profiling were observed, and the degree of conversion ranged from 57.36% (±2.50%) for G 2% to 61.88% (±1.91%) for G 0.5% , without a statistically significant difference among groups (p > 0.05). The UTS values ranged from 32.85 (±6.08) MPa for G 0.5% to 35.12 (±5.74) MPa for G Ctrl (p > 0.05). MYTAB groups showed antibacterial activity against biofilm formation from 0.5 wt % (p < 0.05) and against planktonic bacteria from 1 wt % (p < 0.05). The higher the MYTAB concentration, the higher the cytotoxic effect, without differences between G Ctrl e G 0.5% (p > 0.05). In conclusion, the addition of 0.5 wt % of MYTAB did not alter the physical and chemical properties of the dental resin and provided antibacterial activity without cytotoxic effect.