This study aimed to assess the mechanical and biocompatibility properties of dental resin reinforced with different nanoparticle additives. Temporary crown specimens were 3D-printed and grouped based on nanoparticle type and amount, including zirconia and glass silica. Flexural strength testing evaluated the material’s ability to withstand mechanical stress using a three-point bending test. Biocompatibility was tested using MTT and dead/live cell assays to assess effects on cell viability and tissue integration. Fractured specimens were analysed using scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) for fracture surface examination and elemental composition determination. Results show that adding 5% glass fillers and 10–20% zirconia nanoparticles significantly improves the flexural strength and biocompatibility of the resin material. Specifically, the addition of 10%, 20% zirconia, and 5% glass silica by weight significantly increases the flexural strength of the 3D-printed resins. Biocompatibility testing reveals cell viabilities greater than 80% in all tested groups. Reinforced 3D-printed resin holds clinical potential for restorative dentistry, as zirconia and glass fillers have been shown to enhance mechanical and biocompatibility properties of dental resin, making it a promising option for dental restorations. The findings of this study may contribute to the development of more effective and durable dental materials.