The purpose of the current study was to evaluate the effect of Diode LLLT 650 nm, TEMPO oxidized Nano-fibrillated cellulose mixed with Nano-Amorphous calcium phosphate, and their combination on bone healing in rabbit tibia using H&E staining and computed tomography. Eighteen adult male New Zealand rabbits were selected, two circular bone defects were created in each tibia, resulting in four bony defects in each rabbit, representing the four tested groups; group A (negative control), group B (filled with mineralized nano-cellulose), group C (combination), group D (laser). Animals were euthanized after two weeks and one month, defects were assessed by CT for bone density, then histological samples were examined by H&E stain. In both evaluation periods, group D recorded the greatest mean area percent of new bone formation and bone density, followed by group A, while group C recorded the lowest value. Groups A and D showed full closure of the defects, while groups B and C showed partial defect closure with retained bone graft material. H&E and CT showed that Laser group had the best results of defects healing, bone density and new bone formation, followed by the negative control group. Diode laser 650nm photobiomodulation significantly improved bone defects healing. Mineralized nano-cellulose experimental bone substitute material showed a delayed effect in bone healing and graft material resorption. The combination of LLLT with the graft material had no positive outcome on bone defect healing.