Running is an exercise that can be performed in different environments that imposes distinct foot-floor interactions. For instance, running on grass may help reducing instantaneous vertical impact loading, while compromising natural speed. Inter-muscular coordination during running is an important factor to understand motor performance, but little is known regarding the impact of running surface hardness on inter-muscular coordination. Therefore, we investigated whether inter-muscular coordination during running is influenced by running surface. Surface electromyography (EMG) from 12 lower limb muscles were recorded from young male individuals (n = 9) while running on grass, concrete, and on a treadmill. Motor modules consisting of weighting coefficients and activation signals were extracted from the multi-muscle EMG datasets representing 50 consecutive running cycles using non-negative matrix factorization. We found that four motor modules were sufficient to represent the EMG from all running surfaces. The inter-subject similarity across muscle weightings was the lowest for running on grass (r = 0.76 ± 0.11) compared to concrete (r = 0.81 ± 0.07) and treadmill (r = 0.78 ± 0.05), but no differences in weighting coefficients were found when analyzing the number of significantly active muscles and residual muscle weightings (p > 0.05). Statistical parametric mapping showed no temporal differences between activation signals across running surfaces (p > 0.05). However, the activation duration (% time above 15% peak activation) was significantly shorter for treadmill running compared to grass and concrete (p < 0.05). These results suggest predominantly similar neuromuscular strategies to control multiple muscles across different running surfaces. However, individual adjustments in inter-muscular coordination are required when coping with softer surfaces or the treadmill's moving belt.
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