Peripheral neuropathy is the most common long-term complication in diabetes and is involved in changes in diabetic gait and posture. The regression of nerve function leads to various deficits in the sensory and motor systems, impairing afferent and efferent pathways in the lower extremities. This study aimed to examine how reduced plantar-afferent feedback impacts the gait pattern. Cutaneous sensation in the soles of both feet was experimentally reduced by means of intradermal injections of an anaesthetic solution, without affecting foot proprioception or muscles. Ten subjects performed level walking at a controlled velocity before and after plantar anaesthesia. Muscle activity of five leg-muscles, co-contraction ratios for the knee and ankle joint, ground reaction forces (GRF), spatiotemporal characteristics, joint angles and moments of the hip, knee and ankle were analysed. The intervention significantly lowered plantar sensation, reducing it to the level of sensory neuropathy. Spatiotemporal gait characteristics remained unchanged. The ankle joint was more dorsiflexed which coincided with increased tibialis anterior and decreased gastrocnemius medialis muscle activity during foot flat to mid-stance. In addition, the knee joint was more flexed accompanied by increased biceps femoris activity and higher internal knee-extension moment. With regard to gait dynamics, a delay of the first peak of the vertical GRF was observed. Increased soleus and tibialis anterior muscle activity were found during the end of stance. Short-term loss of plantar sensation affects lower-limb kinematics and gait dynamics, particularly during the first half of stance, and contributes to modified muscle-activation patterns during locomotion.