The objective of this study was to evaluate the effect of different concentrations of NaOCl with and without passive ultrasonic irrigation (PUI) on mechanical properties of human dentin for applications in regenerative endodontics (RE). Sixty single‐rooted teeth were sectioned into 2 halves (n = 120). Dentin bars were produced from one half for flexural strength and the other half was used for microhardness. Specimens were randomly assigned into 10 groups: G1 and G2 (control): distilled water for 30 and 60 min, respectively; G3: 1.5%NaOCl for 30 min; G4:1.5%NaOCl for 60 min; G5: 1.5%NaOCl + PUI for 30 min; G6: 1.5%NaOCl + PUI for 60 min; G7: 5.25%NaOCl for 30 min; G8: 5.25%NaOCl for 60 min; G9: 5.25%NaOCl + PUI for 30 min; G10: 5.25%NaOCl + PUI for 60 min. An increase in NaOCl concentration showed highly significant reduction in mechanical properties. There was no significant difference between 1.5% NaOCl and control group except for specimens treated with PUI for 60 min. NaOCl in concentrations recommended for RE did not have a significant effect on mechanical properties of dentin. However, PUI with increased irrigation time might have an effect even with low NaOCl concentration.