The short-term effect of cryopreservation on specific mechanical behaviors of bovine articular cartilage has been investigated. A flat-ended nonporous indentor was used in a nondestructive, repetitive, axisymmetric unconstrained testing system. Cyclical indentation from a fixed position to a fixed load was applied until a steady-state load-deformation relationship (limit cycle) was achieved. Indentation behaviors measured from the limit cycles of each articular cartilage specimen before and after treatment were compared. Testing was done in vitro using fresh, mature bovine radiocarpal joints. Twenty pairs of cartilage-subchondral bone cores from anatomically similar sites on contralateral joints were separated into three groups; thickness controls, dimethylsulfoxide (DMSO) controls, and cryopreserved experimental samples. Thickness controls and DMSO controls were used to examine the isolated effects of the thickness measurement and DMSO incubation techniques on articular cartilage indentation characteristics. Experimental samples were cryopreserved using DMSO, their thicknesses similarly measured and indentation behaviors examined. Following testing, histological and histochemical assessment of the specimens confirmed the nondestructive nature of the tests. Intra- and intergroup comparisons of controls and experimentals revealed no statistical differences in the mechanical behaviors measured from the limit cycle or in cartilage thickness. These results indicate that the cryopreservation protocol used did not have an effect that we could measure on these specific mechanical behaviors of articular cartilage.