This proof-of-concept study demonstrates that repetitive loading to the pain threshold can safely recreate overloading-induced soft tissue damage and that localised tissue stiffening can be used as a marker for injury. This concept was demonstrated here for the soft tissue of the sole of the foot where it was found that repeated loading to the pain threshold led to long-lasting statistically significant stiffening in the areas where pressure was most intense. Loading at lower magnitudes did not have the same effect. This method can shed new light on the aetiology of overloading injury in the foot to improve the management of conditions such as diabetic foot ulceration and heel pain syndrome. At the same time, the presented concept can also enable the direct assessment of subject-specific thresholds for overloading in other soft tissues which are sensitive to pain, accessible for imaging and can be loaded in a clinically relevant manner.