This study aimed to assess acute and residual changes in sprint-related hamstring injury (HSI) risk factors after a football (soccer) match, focusing on recovery within the commonly observed 72-h timeframe between elite football matches. We used a multifactorial approach within a football context, incorporating optical and ultrastructural microscopic analysis of BFlh (biceps femoris long head) muscle fibres, along with an examination of BFlh fibre composition. Changes in sprint performance-related factors and HSI modifiable risk factors were examined until 3 days after the match (MD ) in 20 football players. BFlh biopsy specimens were obtained before and at MD in 10 players. The findings indicated that at MD , sprint-related performance and HSI risk factors had not fully recovered, with notable increases in localized BFlh fibre disruptions. Interestingly, match load (both external and internal) did not correlate with changes in sprint performance or HSI risk factors nor with BFlh fibre disruption. Furthermore, our study revealed a balanced distribution of ATPase-based fibre types in BFlh, with type-II fibres associated with sprint performance. Overall, the results suggest that a 72-h recovery period may not be adequate for hamstring muscles in terms of both HSI risk factors and BFlh fibre structure following a football match.