Objective: To compare the efficacy of ProFile File cleaner and a nanostructured foam in removing debris from two rotary nickel-titanium instruments during and after instrumentation. Method and Materials: Twenty mandibular premolars were decoronated and instrumented up to either #40.04 RaCe (#40.04) or K3 (#40.04) instruments (n = 10). Each group of files was randomly divided into two groups: ProFile File cleaner and Nano Foam cleaner. The instruments were examined under a scanning electron microscope (SEM) before and after placement into the canals. Subsequently, instruments were introduced into each cleaner and examined again under SEM. The amount of debris removal from the instruments was determined by ImageJ software. Data were subjected to Kruskal-Wallis and Mann-Whitney tests at a .05 level of significance. Results: The amount of debris removal was significantly different between the groups (P = .002). The amount of debris removed was significantly higher in K3 with Nano Foam cleaner than ProFile File cleaner (P = .009). Conclusion: Nanostructured foam can remove debris from endodontic files, especially from those with complex cross-section. This method is recommended to help optimize debris removal with the further possibility of instrument sterilization and reduced risk of cross-contamination. Keywords: debris removal, nanostructured foam, ProFile File cleaner, rotary nickel-titanium instrument, scanning electron microscope