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This research is to investigate the effect of 810 nm diode laser irradiation root dentine mineral contents and smear layer removal when used along with 2.5% (NaOCl) and 17% (EDTA) irrigation. Eighteen (18) freshly extracted permanent premolar teeth were collected; the sample was chemo-mechanically prepared using ProTaper rotary nickel-titanium ((Ni-Ti) instruments and 2.5% (NaOCl). The Sample was divided into, Group A: (8) teeth treated with 5 ml of 17% EDTA only for 1 minute, Group B: (8) teeth were treated with 5 ml of 17% EDTA A for 1 minute then irradiation with 2 W power continuous wave of 810 nm diode laser. A control group of (2) teeth were irrigated only with 5 ml saline. Teeth were then sectioned, (1000x and 2000x) photomicrographs of the root canal walls were taken at 2 mm, 7 mm, and 12 mm depth from the apex the examined under SEM. Regarding smear layer removal no statistically significant difference was detected between the two groups (P>0.05) in term of cracks and conjugation less changes were observed in teeth treated with EDTA a lone compared with Diode +EDTA group (12.5% & 21% respectively). 810 nm diode laser irradiation of root canal dentin irrigation has no significant effect on removal of smear layer. It can cause more dentine erosion with slightly more additional loss of mineral content when used along with NaOCl and EDTA. [ABSTRACT FROM AUTHOR]