Tibial defect, or fracture is very routine musculoskeletal case which brings fully uncomfortable and painful situations to patient. Moreover, it has long hospitalization period because of its risk of non-union. There are many studies using ultrasound, vibration, and laser for bone regeneration to figure out fast bone healing. Among them, Low Level Laser Therapy (LLLT) is already known that it is very easy to treat and may have positive effect for bone regeneration. However, LLLT has uncertain energy dose because of scattering and absorption of laser in tissue. In this study, we used interstitial LLLT to treat tibial defect in animal study. The Interstitial LLLT can overcome some limitations caused by laser scattering or absorption in tissue medium. The results were evaluated using u-CT which can calculate X-ray attenuation coefficient and bone volume of bone defect area. These results showed that interstitial LLLT may affect fast bone healing process in early phase.