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The purpose of this study was to investigate short-term biomechanical changes during treadmill running and whether footwear influenced this adaptation. Forty-five adults with experience in treadmill running over the last year performed an 8.5-min trial at a self-selected speed with three different midsole hardness. Kinetics, kinematics, soft-tissue vibrations and electromyographic activity were recorded at minutes 0, 2, 4, 6 and 8. The fastest adjustments were an increase of active peak and foot inversion. Step frequency and lower limb stiffness decreased then plateaued at minute 6. Duty factor, contact and flight times didn’t reach a plateau. No time effect was found for passive peak, loading rate, leg muscle activity and soft-tissue vibrations. We recommend a minimum habituation period of 8-minutes to ensure that a maximum of biomechanical parameters reach a steady state.