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The present work aims at studying the effect of cortical bone mineralization on ultrasound axial transmission (UAT) measurements, using experiments and numerical simulations. To this end, a reverse fracture healing approach is proposed, using an in vitro bovine femur sample. A demineralization protocol was applied, and the time-of-flight of the wave propagating in axial transmission measured. Numerical simulations of the experimental situation were also implemented. Mineralization degree may influence ultrasound measurements, and it may be a promising tool for fracture healing assessment.