In vivo characterization of Achilles subtendon function and morphology within the tendon cross section and along the free tendon

Abstract

The Achilles tendon is composed of three distinct fascicle bundles, or "subtendons," each originating from the head of one of the three triceps surae muscles. In a healthy tendon, these subtendons slide relative to each other during muscle contractions. This subtendon sliding is reduced in older adults and younger adults who suffer an Achilles tendon injury. However, subtendon sliding is challenging to quantify in low-load scenarios that are critical for monitoring subtendon biomechanics in patients with mechanically compromised tendons, like following an Achilles tendon rupture and repair surgery. The purpose of this study was to develop a reliable method to characterize subtendon behavior in vivo using combined transverse plane ultrasound imaging and neuromuscular electrical stimulation of individual gastrocnemii. We used a Kanade-Lucas-Tomasi point tracking algorithm to quantify tendon displacement during isolated muscle stimulations. Next, we applied k-means clustering to characterize heterogeneous subtendon behavior within the tendon cross section. The tendon cross section displayed differential displacement patterns depending on the stimulated muscle (p<0.0001), and these displacements differed along the free tendon during lateral gastrocnemius stimulations (p=0.004). These results reflect possible differences in load-sharing between adjacent subtendons and differing muscle-tendon dynamics among the triceps surae muscles. Finally, this method confirmed bilaterally symmetric subtendon behavior and demonstrated high inter-session reliability (ICC>0.83). Overall, this study furthers our understanding of differential muscle-tendon dynamics of individual Achilles subtendons both within the tendon cross section and along the free tendon. Future work will apply this method to injured populations to develop biomarkers of altered subtendon function.

Keywords: Achilles Subtendons; muscular stimulation; ultrasound.

Figure 1.

Experimental setup. A) Placement of NMES stimulation electrodes and EMG recording electrodes. B) Ultrasound imaging setup with ankle in a neutral position and transverse ultrasound imaging at five locations along the free tendon. Vector fields overlaid on ultrasound images of the Achilles tendon cross section demonstrate heterogeneous responses of the tendon to NMES along the free tendon. C) Point tracking and k-means clustering workflow. D) Example mean trajectories of points within the identified clusters. Displacement values rapidly increase upon stimulation onset and rapidly decrease after the stimulation ends and the tissue relaxes.

Figure 2.

Mean direction of peak displacement of clusters during GM and GL stimulations. *p<0.05, ***p<0.001, ****p<0.0001.

Figure 3.

Mean cumulative displacement of points during A) GL stimulations and B) GM stimulations. **p<0.01, ***p<0.001.

Figure 4.

A) Medial-lateral position of the centroid of the most displaced cluster during GM and GL stimulations. B) Anterior-posterior position of the centroid of the most displaced cluster during GM and GL stimulations. C-D) Separate views of the cluster medial-lateral positions during GM and GL stimulations. E-F) Separate views of the cluster anterior-posterior positions during GM and GL stimulations. *p<0.05, **p<0.01, ***p<0.001, ***p<0.0001.

Figure 5.

Area of the cluster representing the most displaced region within the tendon cross section, represented as a fraction of the tendon cross sectional area, measured at five locations along the free tendon during both GM and GL stimulations. *p<0.05.

Figure 6.

Achilles tendon cross-sectional area, width, and thickness did not differ between right and left legs.

Figure 7.

Measurements of cluster centroid positions and area fractions did not differ between right and left legs. Panels A, B, and C display bilateral comparisons of cluster medial-lateral position, anterior-posterior position, and cluster area fraction, respectively, during GL stimulations. Panels D, E, and F display bilateral comparisons of cluster medial-lateral position, anterior-posterior position, and cluster area fraction, respectively, during GM stimulations.