The current study reports the sensitivity of acoustical parameters estimated at high frequency to the osteoarthritic morphological and structural changes in patellar cartilage in rat knees. Osteoarthritis (OA) was induced by a single intra-articular injection of mono-iodo-acetic acid in right knees. OA patellas and their contralateral controls were excised at regular intervals after injection and were examined in vitro with a scanning acoustical microscope operating with a poly(vinylidene di-fluoride) (PVDF) 80-MHz focused transducer. Cartilage thickness was estimated using B-scan images. The quantitative analysis of the radiofrequency signal backscattered by the cartilage was performed using integrated reflection coefficient (IRC) and apparent integrated backscatter (AIB), which were estimated in the 20-60-MHz frequency range. One week after injection, a cartilage thickness decrease was detected (-6%, on average) that preceded the significant hypertrophy (20.1%) that occurred 2 weeks after injection and could be due to tissue repair. From 1 week to 3 weeks after injection, the IRC of OA patellas was significantly lower than that of control patellas. The IRC difference increased with time from -3.3 +/- 2.4 dB at 1 week to -8.4 +/- 1.7 dB at 3 weeks. An AIB decrease was observed with time for both OA and control patellas (-2.9 to -4.2 dB per week). An AIB difference between OA and control patellas was detected from 1 week to 3 weeks after injection. This difference decreased with time. IRC variation reflects a change in acoustical impedance of the superficial layer of the cartilage and could be linked to a change in constituent content and/or to a disruption of fibers of the collagen network that led to the fibrillation of the cartilage surface. AIB variation reflects a change in shape, size and/or density of the scatterers and could be related to changes in the constituent content and in the organization of the matrix in the internal layer of the cartilage. IRC and AIB could provide information about the structural modifications of the cartilage due to osteoarthritis or to cartilage maturation.