A significant problem related to the functioning of resin-based composites for dental fillings is secondary or recurrent caries, which is the reason for the need for repeated treatment. The cross-linked quaternary ammonium polyethylenimine nanoparticles (QA-PEI-NPs) have been shown to be a promising antibacterial agent against different bacteria, including cariogenic ones. However, little is known about the properties of dental dimethacrylate polymer-based composites enriched with QA-PEI-NPs. This research was carried out on experimental composites based on bis-GMA/UDMA/TEGDMA matrix enriched with 0.5, 1, 1.5, 2 and 3 (wt%) QA-PEI-NPs and reinforced with two glass fillers. The cured composites were tested for their adherence of Streptococcus Mutans bacteria, cell viability (MTT assay) with 48 h and 10-days extracts , degree of conversion (DC), water sorption (WSO), and solubility (WSL), water contact angle (CA), flexural modulus (E), flexural strength (FS), compressive strength (CS), and Vickers microhardness (HV). The investigated materials have shown a complete reduction in bacteria adherence and satisfactory biocompatibility. The QA-PEI-NPs additive has no effect on the DC, VH, and E values. QA-PEI-NPs increased the CA (a favorable change), the WSO and WSL (unfavorable changes) and decreased flexural strength, and compressive strength (unfavorable changes). The changes mentioned were insignificant and acceptable for most composites, excluding the highest antibacterial filler content. Probably the reason for the deterioration of some properties was low compatibility between filler particles and the matrix; therefore, it is worth extending the research by surface modification of QA-PEI-NPs to achieve the optimum performance characteristics.