Understanding how loading and damage on common running injury locations changes across speeds, surface gradients, and step frequencies may inform training programs and help guide progression/rehabilitation after injuries. However, research investigating tissue loading and damage in running is limited and fragmented across different studies, thereby impairing comparison between conditions and injury locations. This study examined per‐step peak load and impulse, cumulative impulse, and cumulative weighted impulse (hereafter referred to as cumulative damage) on three common injury locations (patellofemoral joint, tibia, and Achilles tendon) across different speeds, surface gradients, and cadences. We also explored how cumulative damage in the different tissues changed across conditions relative to each other. Nineteen runners ran at five speeds (2.78, 3.0, 3.33, 4.0, 5.0 m s−1), and four gradients (−6, −3, +3, +6°), and three cadences (preferred, ±10 steps min−1) each at one speed. Patellofemoral, tibial, and Achilles tendon loading and damage were estimated from kinematic and kinetic data and compared between conditions using a linear mixed model. Increases in running speed increased patellofemoral cumulative damage, with nonsignificant increases for the tibia and Achilles tendon. Increases in cadence reduced damage to all tissues. Uphill running increased tibial and Achilles tendon, but decreased patellofemoral damage, while downhill running showed the reverse pattern. Per‐step and cumulative loading, and cumulative loading and cumulative damage indices diverged across conditions. Moreover, changes in running speed, surface gradient, and step frequency lead to disproportional changes in relative cumulative damage on different structures. Methodological and practical implications for researchers and practitioners are discussed.