© 2021, The Society of The Nippon Dental University.The aim of this study was to evaluate the antibacterial efficacy of copper added chitosan nanoparticles (CU-CNPs) as an irrigation solution with different irrigants in terms of eliminating Enterococcus Faecalis (E. faecalis) from the root canals. Fifty mandibular premolar teeth were prepared and infected with E. faecalis for 21 days. After the incubation period, samples were randomly divided into a control group irrigated with distilled water and 4 experimental groups (n = 10) irrigated with as follows, %6 NaOCl, %6 NaOCl + %9 editronate (HEBP), Chitosan nanoparticles (CNPs), and CU-CNPs. To calculate the proportion of dead E. faecalis cell volume, stained using LIVE/DEAD BacLight Bacterial Viability Kit and were scanned using confocal laser scanning microscope (CLSM). All the irrigation solutions significantly (P <.05) killed the bacteria in the canal, except for the control group. CU-CNPs solution killed the highest (P <.05) number of bacteria compared with the other experimental groups. No significant difference was found between CNPs, NaOCl + HEBP, and NaOCl in terms of antibacterial activity. CU-CNPs solution was exhibited higher antibacterial efficacy against E. faecalis.