Water content alters viscoelastic behaviour of the normal adolescent rabbit medial collateral ligament

Testing environment is an important factor in the outcome of mechanical tests on connective tissue. The purpose of this investigation was to determine the effect of ligament water content on ligament mechanical behaviour by altering the test environment. Water content of medial collateral ligament (MCLs) from 19 three-month-old New Zealand White rabbits was varied in subsets of ligaments pairs by means of immersion in 2, 10 or 25% sucrose or 0.9% phosphate-buffered saline (PBS) solutions for 1 h. One knee joint was cycled 50 times in the designated solution (experimental), while the contralateral knee (uncycled control) was simultaneously soaked in the same tank. Following cycling, the water contents of both test and control ligaments were determined. Water contents of 22 normal MCLs were determined immediately post-sacrifice and served as 'normal water content' controls. Normalized peak cyclic load changes were used as a measure of the viscoelastic behaviour of each MCL. Results demonstrated that only ligaments soaked (but not cycled) in a 10% sucrose solution had water contents (60.5 +/- 2.5%) which were statistically similar to the 22 fresh normal MCLs (63.9 +/- 6.0%). Ligaments soaked in PBS (74.0 +/- 1.3%) or 2% sucrose (69.2 +/- 2.3%) had significantly higher water contents compared to fresh normal MCLs. Ligaments with higher water contents (e.g. soaked in PBS or 2% sucrose) demonstrated greater cyclic load relaxation compared to ligaments with lower contents (e.g. soaked in 25 or 10% sucrose). Different fluid test environments can significantly alter ligament water content and, in turn, significantly affect ligament viscoelastic behaviour.