Introduction: Recently, foam rolling has become an increasingly popular technique for the relaxation of muscle tissue in professional football players who participate in demanding training schedules. The mechanism behind foam rolling is not fully understood. Purpose: The purpose of the study was to determine the effect of foam rolling on the contractile properties of the knee muscles in football players, after a sports-specific load. Methods: In a controlled study, we included 20 male professional football players and measured the contractile properties of biceps femoris, rectus femoris, vastus medialis and vastus lateralis muscles on the dominant lower limb with tensiomyography before and after Yo-Yo interval test and after 5 × 45 s of hamstring foam rolling. We also measured active and passive range of motion of knee extension before and after the intervention. A one-way analysis of variance (ANOVA) was performed for normally distributed data, and non-parametric median comparison tests were used for abnormally distributed data. A mixed linear model was performed to determine the differences between the mean values of the groups. The level of statistical significance was determined with p<0.05. Results: There were no statistically significant differences between the experimental and the control group for any of the measured muscles in Tc, Td, and Dm after the Yo-Yo interval test or after the SPV intervention (p>0.05). There were also no statistically significant differences in the active and passive range of motion of extension in the knee joint between the groups. Discussion and conclusion: Five repetitions of 45 seconds of hamstring foam rolling had no statistically significant effect. That is; for the delay time, contraction time, maximum muscle amplitude of the biceps femoris or on the range of motion in professional football players. Many effects associated with foam rolling are probably not affecting the mechanical properties of the muscle itself.