Background and Objectives: In endodontics, various intracanal medications have been advocated to eliminate bacteria after root canal instrumentation. A recent study has revealed that addition of powdered dentin to bioactive glass (BAG) led to increased glass dissolution, and an increased antibacterial efficacy. Therefore, the present study was undertaken to compare the effect of enamel and dentin powder on the antibacterial efficacy of a commercially available BAG. Materials and Methods: Dentin blocks (dbs) were prepared from single rooted human teeth. These dbs were infected with Enterococcus faecalis for two weeks in Tryptic Soy Broth (TSB), while negative controls were kept in sterile TSB. In group I, the infected dbs were filled with BAG, in group II with BAG + Enamel powder and group III with BAG + Dentin powder. Dentin samples were harvested from the dbs and cultured. Statistical Analysis: Kruskal-Wallis ANOVA was used for multiple group comparison followed by Scheffe′s post hoc test for pair-wise comparisons. Results: All the combinations of BAG evaluated significantly reduced the bacterial counts compared to the control group. However, at the end of 24 hours, three days, and five days BAG + Dentin powder showed significant reduction ( P < 0.01) in bacterial counts compared to the other experimental groups. Conclusion: Among the various materials evaluated, it appeared that though BAG exhibits antimicrobial efficacy, the addition of powdered enamel and dentin in aqueous suspension definitely enhanced this property. However, the addition of enamel powder BAG did not significantly alter its antimicrobial efficacy compared to BAG + dentin powder.