Macroscopically healthy articular cartilage with fibrillar-scale early tissue degeneration subject to impact loading results in greater extent of cell-death

From previous investigations it has been shown that there exists healthy-appearing articular cartilage that contains collagen fibril network destructuring. It is hypothesised that such sub-micron scale destructuring not only presents an increased vulnerability to tissue scale damage following impact loading, but an increase in cell death as well. Cartilage-on-bone blocks from 12 patellae, six healthy (G0) and the other six with sub-micron fibrillar destructuring (G1), were obtained and subject to 2.3 J impact loading. Two sets of sub-samples were obtained for each block tested. One set was used to examine for the live/dead cell response using calcein-AM and propidium iodide staining, imaged with confocal microscopy. The tissue microstructural matrix was imaged from the other matched set, unstained and in its fully hydrated state, using differential interference contrast optical light mi-croscopy. High speed imaging of the impact was used to calculate the velocity changes or coefficient of restitution (COR) and used as a proxy of energy that the tissue absorbed. A previously defined tissue matrix damage score was used to quantify the extent of fracturing and cracking in the matrix. The cell death (PCD) was counted and presented as a percentage against all cells live plus dead. The energy absorbed was 36.5% higher in G1 than in G0 (p = 0.034). However, the damage score and PCD of samples in the G1 group was much larger than the G0 group, similar to 300% and 161% respectively. Microscopy showed that cell death is associated to both matrix compaction and further fibrillar destructuring from the ECM to the territorial matrix regions of the chondron. Following impact loading, cartilage tissue that appears normal but contains sub-micron fibrillar matrix destructuring responds with significantly increased cell death.