Isolated injury to the long head of biceps femoris is the most common type of acute hamstring strain injury (HSI). However, the precise hamstring injury mechanism (i.e., sprint-type) is still not well understood, and research is inconclusive as to which phase in the running cycle HSI risk is the greatest. Since detailed information relating to hamstring muscle function during sprint running cannot be obtained in vivo in humans, the findings of studies investigating HSI mechanisms are based on modeling that requires assumptions to be made based on extrapolations from anatomical and biomechanical investigations. As it is extremely difficult to account for all aspects of muscle-tendon tissues that influence function during high-intensity running actions, much of this complexity is not included in these models. Furthermore, the majority of analyses do not consider the influence of prior activity or muscular fatigue on kinematics, kinetics and muscle activation during sprinting. Yet, it has been shown that fatigue can lead to alterations in neuromuscular coordination patterns that could potentially increase injury risk. The present critical review will evaluate the current evidence on hamstring injury mechanism(s) during high-intensity running and discuss the interactions between fatigue and hamstring muscle activation and function.