Background: Clinically, three-dimensional (3D) printing technology is becoming a popular and efficient dental processing technology. Recently, there has been an increasing demand for dental materials that exhibit bioactive properties. The present study aimed to evaluate the mechanical properties, cytotoxicity, and fluoride ion release capacity of 3D-printed dental resins modified with bioactive glass. Materials and methods: The resin FotoDent splint used in the production of removable orthodontic splints, was modified by the addition of two types of bioactive glasses that are capable of releasing fluoride ions. The novel materials used for the production of dental splints were examined for their mechanical, physical, and biological properties (fracture resistance, sorption, solubility, elution of nonpolymeric substances, and release of fluoride ions over time) and cytotoxic effects on cell cultures. Results: Initially, the fracture toughness of the 3D-printed resin was found to be 55 MPa, but after modification with glass, the resistance was reduced to about 50 MPa. Sorption and solubility values of the materials (19.01 ÷ 21.23 µg/mm3 and 0.42 ÷ 1.12 µg/mm3, respectively) complied with the safety limits imposed by ISO standard. Modified resins were capable of releasing fluoride ions, and the maximum releasing effect was observed after 14 days of incubation. Both the modified resins, after four days of contact with human gingival fibroblasts, exhibited moderate cytotoxic properties. Conclusions: The experimental results showed that modification of methacrylate resin, used in 3D printing technology, with bioactive glasses produces novel dental materials that possess desirable bioactive properties. The findings of this study indicate the potential ability of modified polymethacrylate resins to release fluoride ions in the oral cavity environment. The modified materials are characterized with a moderate decrease in physical properties and mild cytotoxicity on direct contact with human fibroblasts.