AbstractMost studies on articular cartilage properties have been conducted after detachment of the cartilage from the bone. In the present work we investigated the effect of detachment on collagen fiber architecture. We used one‐dimensional 2 H double quantum filtered MRI on cartilage bone plugs equilibrated in deuterated saline. The quadrupolar splittings observed in the different zones were related to the degree of order and the density of the collagen fibers. The method is non‐destructive, allowing for measurements on the same plug without the need for fixation, dehydration, sectioning and decalcification. Detachment of the radial from the calcified zone resulted in swelling of the cartilage plug in physiological saline and a concomitant decrease in the quadrupolar splitting. The effect of mechanical pressure on the 2 H quadrupolar splittings for the detached cartilage and for the calcified zone‐bone plugs were compared with those of the same zones in the intact cartilage–bone plug. The splitting in the radial zone of the detached cartilage collapsed at much smaller loads compared to the intact cartilage–bone plug. The effect of the load on the size of the cartilage was also greater for the detached plug. These results indicate that anchoring of the cartilage to the bone through the calcified zone plays an important role in retaining the order of the collagen fibers. The water 2 H quadrupolar splitting in intact and proteoglycan‐depleted cartilage was the same, indicating that the proteoglycans do not contribute to the ordering of the collagen fibers. © 2004 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.