Ex-vivo validation of spatial gain sonography for the quantification of echo intensity in fascicle-aligned ultrasound images in ten anatomical muscles in Bos taurus

This study aimed to validate the concept of spatial gain sonography for quantifying texture-related echo intensity in B-mode ultrasound of skeletal muscle. Fifty-one bovine muscles were scanned postmortem using B-mode ultrasonography at varying fascicle probe angles (FPA). The relationship between mean gray values (MGV) and FPA was fitted with a sinusoidal and a linear function, the slope of which was defined as tilt echo gain (TEG). Macroscopic muscle cross sections were optically analyzed for intramuscular connective tissue (IMCT) content which was plotted against MGV at 0° FPA (MGV_00). MGV peaked at FPA 0°. Sine fits were superior to linear fits (adjusted r 2 -values 0.647 vs. 0.613), especially for larger FPAs. In mixed models, the pennation angle was related to TEG ( P < 0.001) and MGV_00 ( P = 0.035). Age was relevant for MGV_00 ( P < 0.001), but not TEG ( P > 0.10). The correlation between the IMCT percentage and MGV_00 was significant but weak ( P = 0.026; adjusted r 2 = 0.103). The relationship between fascicle probe angle and echo intensity in B-mode ultrasound can be modeled more accurately with a sinusoidal but more practically for clinical use with a linear fit. The peak mean gray value MGV_00 can be used to compare echo intensity across muscles without the bias of pennation angle.