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Introduction: The successful outcome of endodontic retreatment heavily relies on the thorough removal of root canal filling materials. This study aimed to quantitatively and qualitatively assess the efficacy of various retreatment file systems using nano-computed tomography (nano-CT) imaging. Materials and Methods: Thirty recently extracted human mandibular molars with mesial roots having curvatures <20° were chosen and sectioned to 16 mm in length. The root canals were prepared with the ProTaper Gold system up to size F2 and they were filled with gutta percha and AH Plus sealer. After obturation, all the samples were randomly assigned to two groups (n = 15 each) for retreatment: Group 1 utilized Solite RE Black retreatment files, while Group 2 employed Coltene HyFlex Remover files. Nano-CT imaging was employed to measure pre- and postoperative volumes of remaining filling material and remaining dentin thickness and the time taken for retreatment was measured in seconds. The obtained values were analyzed statistically (P < 0.05). Results: Nano-CT analysis showed a significant difference (P < 0.05) in the volume of remaining filling material between the Solite RE Black and Coltene HyFlex Remover groups. However, no significant difference was observed in the time required for gutta-percha removal. The Solite RE Black system showed comparatively less dentin removal than the Coltene HyFlex Remover system (P < 0.05). Conclusion: The Solite RE Black retreatment file system demonstrated promising efficacy in removing filling material while preserving dentin integrity compared to the Coltene HyFlex Remover system. These findings support its potential as an effective tool for endodontic retreatment, emphasizing the need for further clinical trials to validate its performance across various clinical scenarios. [ABSTRACT FROM AUTHOR]