OBJECTIVE: To evaluate the normal location of the peroneus longus tendon (PL) in the cuboid groove in various ankle-foot positions by ultrasonography in asymptomatic volunteers.
METHODS: Ultrasonographic assessment of the PL in the cuboid groove was performed in 20 feet of ten healthy volunteers. Each PL was examined in five ankle-foot positions (i.e., neutral, dorsiflexion, plantar-flexion, supination, and pronation). The PL location was qualitatively categorized as \"inside\" when the PL was entirely within the cuboid groove, as \"overlying\" when some part of the PL was perched on the cuboid tuberosity, and as \"outside\" when the PL was entirely on the cuboid tuberosity. For quantitative evaluation of the PL location, the distance between the PL and the cuboid groove was measured. The width of the cuboid groove was measured in the neutral position.
RESULTS: The PL location did not significantly change with changes in the ankle-foot position. Qualitatively, an \"overlying\" PL was the most common type, regardless of the ankle-foot position. \"Inside\" PLs were found in only 35, 20, 30, 25, and 35% of feet in neutral, dorsiflexion, plantar-flexion, supination, and pronation positions, respectively. The quantitative PL location was also not significantly different among all ankle-foot positions and it was significantly negatively correlated with the cuboid groove width.
CONCLUSIONS: In healthy volunteers, 65% or more of the PLs were partially or completely located outside of the cuboid groove, regardless of the ankle-foot position. The PL location relative to the cuboid groove was related to the cuboid groove width.

The peroneocuboid joint, between the peroneus longus tendon and the cuboid bone, has not been anatomically well-defined and no embryological study has been published. Furthermore, the ossification of the os peroneum (a sesamoid inside the peroneus longus tendon) and its associated pathology has been considered to be generated by orthostatic and/or mechanical loads. A light microscopy analysis of serially sectioned human embryonic and fetal feet, the analysis of human adult feet by means of standard macroscopic dissection, X-ray and histological techniques have been carried out. The peroneus longus tendon was fully visible until its insertion in the 1st metatarsal bone already at embryonic stage 23 (56-57 days). The peroneocuboid joint cavity appeared at the transition of the embryonic to the fetal period (8-9th week of gestation) and was independent of the proximal synovial sheath. The joint cavity extended from the level of the calcaneocuboid joint all the way to the insertion of the peroneus longus tendon in the 1st metatarsal bone. The frenular ligaments, fixing the peroneus longus tendon to the 5th metatarsal bone or the long calcaneocuboid ligament, developed in the embryonic period. The peroneus longus tendon presented a thickening in the area surrounding the cuboid bone as early as the fetal period. This thickening may be considered the precursor of the os peroneum and was similar in shape and in size relation to the tendon, to the os peroneum observed in adults. To the best of our knowledge, this is the first study to show that the os peroneum, articular facets of the peroneus longus tendon and cuboid bone, the peroneocuboid joint and the frenular ligaments appear during the embryonic/fetal development period and therefore they can not be generated exclusively by orthostatic and mechanical forces or pathological processes.