The objective of this work was to determine the antimicrobial and antibiofilm properties of mineral trioxide aggregate (MTA) supplemented with bismuth lipophilic nanoparticles (BisBAL NPs). The antimicrobial activity of the composite MTA-BisBAL NPs was determined by the disk diffusion assay, while antibiofilm activity was analyzed by fluorescence microscopy. The cytotoxicity of MTA-BisBAL NPs was determined on human gingival fibroblasts by optical microscopy and crystal violet staining. MTA-BisBAL NPs inhibited the growth of Enterococcus faecalis, Escherichia coli, and Candida albicans and also detached the biofilm of fluorescent E. faecalis after 24 h of treatment. The addition of BisBAL nanoparticles did not significantly modify the physical properties of MTA, and cytotoxicity was not observed when MTA-BisBAL NPs was added on human gingival fibroblasts. Altogether these results suggest that BisBAL nanoparticles provide antimicrobial and antibiofilm activities to MTA while it retained their biophysical properties without cause side effects on human gingival fibroblasts.