Background: Enamel demineralization and white spot lesions around orthodontic brackets is a common problem associated with fixed orthodontic treatment. Glass ionomer cements (GICs) are commonly used for bracket bonding and recently, bioactive glass (BAG) was added to GICs to enhance deposition of calcium phosphate in adjacent dental structure. This study sought to assess the effect of addition of BAG particles to resin modified GIC (RMGIC) on enamel demineralization under orthodontic brackets. Material and Methods: In this in vitro, experimental study, 60 sound human premolars were immersed in distilled water at 6°C. The teeth were randomly divided into three groups. Using a sticker, a window measuring 7x7 mm was created on the buccal surface. The remaining surfaces were coated with nail varnish twice with a 3-hour interval. After 24 hours, the stickers were removed. Brackets were bonded to tooth surfaces using Transbond XT (control), Fuji II LC RMGIC and Fuji II LC containing 30% BAG particles. To induce acid attacks, the teeth were immersed in demineralizing solution for 6 hours. For pH cycling, the teeth were then immersed in remineralizing solution for 18 hours. This process was repeated for 21 days. Next, the teeth were longitudinally sectioned into two halves and the depth of demineralization was measured from the deepest point to the surface under a polarized light microscope. Data were analyzed using one-way ANOVA, and pairwise comparisons were carried out using Tukey?s test. Results: The mean depth of demineralization was 73.8±22.29, 118.08±29.42 and 182.98±20.69 µm in the BAG, RMGIC and Transbond XT groups, respectively. One-way ANOVA showed significant differences in terms of depth of demineralization among the three groups (P=0.0001). Tukey?s test revealed significant differences in depth of demineralization. Conclusions: Addition of BAG to RMGIC can significantly decrease the depth of enamel demineralization under orthodontic brackets in vitro.