In the past years, there have been several experimental studies that aimed at quantifying the material properties of articular ligaments such as tangent modulus, tensile strength, and ultimate strain. Little has been done to describe their response to mechanical stimuli that lead to damage. The purpose of this experimental study was to characterize strain-induced damage in medial collateral ligaments (MCLs). Displacement-controlled tensile tests were performed on 30 MCLs harvested from Sprague Dawley rats. Each ligament was monotonically pulled to several increasing levels of displacement until complete failure occurred. The stress–strain data collected from the mechanical tests were analyzed to determine the onset of damage and its evolution. Unrecoverable changes such as increase in ligament's elongation at preload and decrease in the tangent modulus of the linear region of the stress–strain curves indicated the occurrence of damage. Interestingly, these changes were found to appear at two significantly different threshold strains (). The mean threshold strain that determined the increase in ligament's elongation at preload was found to be 2.84% (standard deviation (SD) = 1.29%) and the mean threshold strain that caused the decrease in the tangent modulus of the linear region was computed to be 5.51% (SD = 2.10%), respectively. The findings of this study suggest that the damage mechanisms associated with the increase in ligament's elongation at preload and decrease in the tangent modulus of the linear region in the stress–strain curves in MCLs are likely different.