Background: The aim of this in vitro study was to evaluate the effect of different endodontic irrigants on the micro-tensile bond strength (µTBS) of a one-step self-etch adhesive system to dentin surfaces. Material and methods: Freshly extracted human incisor teeth were used as the study sample, with a total of 45 teeth included. The selection criteria excluded hypo plastic and hypo mineralized teeth, teeth with restorations, and teeth with crown fractures. A total of 45 intact human molars were selected and divided into three groups based on the irrigants used. A self-etch universal adhesive system was utilized for composite restoration. In the control group (Group 1), the dentine surface was irrigated with sodium hypochlorite (NaOCl) followed by EDTA. Group 2 received irrigation with sodium hypochlorite followed by 18% HEDP, while in Group 3, sodium hypochlorite was followed by grape seed extract as the irrigant. Resin composites were built up using the Tetric N-Bond Universal adhesive system. The micro tensile bond strength was evaluated using a Universal Testing Machine. Data analysis was done using SSPS software. Results: Group 1 exhibited a mean bond strength of 31.2 ± 2.10 MPa, with a 95% confidence interval (CI) ranging from 22.5 to 24.8. Group 2 had a mean bond strength of 28.6 ± 2.70 MPa, with a 95% CI of 29.5 to 32.4. Group 3 demonstrated the highest bond strength, with a mean value of 33.2 ± 3.13 MPa and a 95% CI of 31.7 to 33.5. A statistically significant difference was observed (p < 0.002), indicating that the bond strength varied significantly among the groups. The confidence interval values provide an estimate of the range within which the true mean bond strength is expected to lie. A significant difference was observed between Group 1 and Group 2 (mean difference: 6.5, p < 0.002) and between Group 1 and Group 3 (mean difference: 7.9, p < 0.003), indicating a statistically significant variation in bond strength. However, the comparison between Group 2 and Group 3 (mean difference: 3.7, p = 0.121) did not show a statistically significant difference. These findings suggest that the bond strength differs notably between certain groups, with a statistically significant threshold set at p < 0.05. Conclusion: The study revealed significant variations in bond strength among the groups, with Group 3 showing the highest values. Statistically significant differences were observed between Group 1 and the other groups, while Group 2 and Group 3 showed no significant difference.