The biostimulatory effects of laser irradiation focus not only in the field of soft tissue but also bone formation. Studies have shown that the light of a nanosecond pulsed laser which has a high peak power can produce stress waves in tissue. We have hypothesized that nanosecond pulsed laser irradiation stimulates bone formation. Our aim was to clarify the mechanism of increased bone volume by nanosecond pulsed laser irradiation. Rat femur was irradiated with a Q-switched Nd:YAG laser, which has a wavelength of 1064 nm. The quantification of trabecular architecture using three-dimensional morphometric analysis and measurement of bone mineral density (BMD) using pQCT was performed on day 1, day 3, day 5, and day 7 after laser irradiation. The laser effects on bone cells were also investigated using histological and immunohistochemical analysis. On day 1 after laser irradiation, bone volume (BV/TV), trabecular thickness (Tb.Th), and other parameters of the irradiated group did not significantly differ from the non-irradiation group (control). However, the mean BV/TV, Tb.Th, mineral apposition rate, and BMD of the laser group on day 7 after laser irradiation were significantly greater than those of the control. On histological analysis, the number of TRAP-positive osteoclasts was lower on day 3 after laser irradiation. Osteoblasts with activated clearance were seen in the laser irradiated group on day 1 and day 3. These data reveal that the increased bone volume by nanosecond pulsed laser irradiation causes an increase in osteoblast activity and a decrease in osteoclast number.