This study was performed to explore the tissue-level changes in mineralization caused by low-intensity ultrasound stimulation after distraction osteogenesis. Unilateral femoral lengthenings (7 mm) were performed on 34 male Sprague-Dawley rats. Half of the animals received daily ultrasound stimulation for 5 weeks; the remaining animals received sham treatments. Healing was assessed with serial radiographs, quantitative micro-computed tomography, and biomechanical testing. Twenty-one animals were evaluated at the conclusion of the study (9 experimental, 12 control). Radiographically, healing of the ultrasound-treated bones preceded that of the sham-treated bones by approximately 1 week. Bone volume fraction and trabecular bone pattern factor were significantly higher in the ultrasound-treated animals, but there were no significant differences in bone mineral content or bone mineral density. The ultrasound-treated femurs were 20% stiffer and 33% stronger than the control femurs, but the differences were not statistically significant. These findings suggest that pulsed, low-intensity ultrasound matures the regenerate by altering the microarchitecture of the newly formed bone.