Aim: The present study was conducted for comparing the efficacy of three root canal sealers: AH Plus, MTA Fillapex, and Bio-C® Sealer. Materials and methods: Root canal sealers are essential components in endodontic therapy, significantly affecting both sealing efficacy and biocompatibility. This study involved thirty freshly extracted mandibular lateral incisors characterized by straight, single root canals. The prepared specimens were subsequently categorized into three groups, each comprising 10 teeth, for obturation with different sealers: Bio C Sealer, AH Plus, and MTA Fillapex. All sealers were mixed in accordance with the manufacturer's instructions. The specimens were then sectioned, and the sealing ability was assessed. Data analysis was performed using SPSS software, with the ANOVA test employed to determine the significance level. Results: Conclusion: Root canal sealers are essential in endodontic therapy, as they significantly affect both the sealing capacity and biocompatibility of the treatment. Notable variations were identified in the sealing effectiveness among AH Plus, MTA Fillapex, and Bio-C Sealers.