| Summary: | <i>Background</i>. Muscle synergy analysis is an approach to understand the neurophysiological mechanisms behind the hypothesized ability of the Central Nervous System (CNS) to reduce the dimensionality of muscle control. The muscle synergy approach is also used to evaluate motor recovery and the evolution of the patients’ motor performance both in single-session and longitudinal studies. Synergy-based assessments are subject to various sources of variability: natural trial-by-trial variability of performed movements, intrinsic characteristics of subjects that change over time (e.g., recovery, adaptation, exercise, etc.), as well as experimental factors such as different electrode positioning. These sources of variability need to be quantified in order to resolve challenges for the application of muscle synergies in clinical environments. The objective of this study is to analyze the stability and similarity of extracted muscle synergies under the effect of factors that may induce variability, including inter- and intrasession variability within subjects and intersubject variability differentiation. The analysis was performed using the comprehensive, publicly available hand grasp NinaPro Database, featuring surface electromyography (EMG) measures from two EMG electrode bracelets. <i>Methods</i>. Intrasession, intersession, and intersubject synergy stability was analyzed using the following measures: variance accounted for (VAF) and number of synergies (NoS) as measures of reconstruction stability quality and cosine similarity for comparison of spatial composition of extracted synergies. Moreover, an approach based on virtual electrode repositioning was applied to shed light on the influence of electrode position on intersession synergy similarity. <i>Results</i>. Intersession synergy similarity was significantly lower with respect to intrasession similarity, both considering coefficient of variation of VAF (approximately 0.2–15% for inter vs. approximately 0.1% to 2.5% for intra, depending on NoS) and coefficient of variation of NoS (approximately 6.5–14.5% for inter vs. approximately 3–3.5% for intra, depending on VAF) as well as synergy similarity (approximately 74–77% for inter vs. approximately 88–94% for intra, depending on the selected VAF). Virtual electrode repositioning revealed that a slightly different electrode position can lower similarity of synergies from the same session and can increase similarity between sessions. Finally, the similarity of intersubject synergies has no significant difference from the similarity of intersession synergies (both on average approximately 84–90% depending on selected VAF). <i>Conclusion</i>. Synergy similarity was lower in intersession conditions with respect to intrasession. This finding should be considered when interpreting results from multi-session assessments. Lastly, electrode positioning might play an important role in the lower similarity of synergies over different sessions.
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