Reliability of assessing skeletal muscle architecture and tissue organization of the gastrocnemius medialis and vastus lateralis muscle using ultrasound and spatial frequency analysis

Introduction: The purpose of this study was to investigate inter- and intra-rater reliability as well as the inter-rater interpretation error of ultrasound measurements assessing skeletal muscle architecture and tissue organization of the gastrocnemius medialis (GM) and vastus lateralis (VL) muscle. Methods: The GM and VL of 13 healthy adults (22 +/- 3 years) were examined thrice with sagittal B-mode ultrasound: intraday test-retest examination by one investigator (intra-rater) and separate examinations by two investigators (inter-rater). Additionally, images from one investigator were analysed by two interpretators (interpretation error). Muscle architecture was assessed by muscle thickness [MT], fascicle length [FL], as well as superior and inferior pennation angle [PA]. Muscle tissue organization was determined by spatial frequency analysis (SFA: peak spatial frequency radius, peak -6 dB width, PSFR/P6, normalized peak value of amplitude spectrum [Amax], power within peak [PWP], peak power percent). Reliability of ultrasound examination and image interpretation are presented as intraclass correlation coefficient (ICC), test-retest variability, standard error of measurement as well as bias and limits of agreement. Results: GM and VL demonstrated excellent ICCs for inter- and intra-rater reliability, along with excellent ICCs for interpretation error of MT (0.91-0.99), showing minimal variability (<5%) and SEM% (<5%). Systematic bias for MT was less than 1 mm. For PA and FL poor to good ICCs for inter- and intra-rater reliability were revealed (0.41-0.90), with moderate variability (<12%), low SEM% (<10%) and systematic bias between 0.1-1.4(degrees). Tissue organization analysis indicated moderate to good ICCs for inter- and intra-rater reliability. Notably, Amax and PWP consistently held the highest ICC values (0.77-0.87) across all analyses but with higher variability (<24%) and SEM% (<18%), compared to lower variability (<9%) and SEM% (<8%) in other tissue organization parameters. Interpretation error of all muscle tissue organization parameters showed excellent ICCs (0.96-0.999) with very low variability (<= 1%) and SEM% (<2%), except Amax & PWP (TRV%: <6%; SEM%: <7%). Conclusion: Our findings demonstrated excellent inter- and intra-rater reliability for MT. However, agreement for PA, FL, and SFA parameters was not as strong. Additionally, MT and all SFA parameters exhibited excellent agreement for inter-rater interpretation error. Therefore, the SFA seems to offer the possibility of objectively and reliably evaluating ultrasound images.