The mechanical significance of the zonally differentiated collagen network of articular cartilage in relation to tissue swelling

Background: We hypothesise that the Benninghoff arcade fibril structure motif of cartilage is able to predict the swelling response of cartilage. Methods: A total of ten healthy adult bovine patellae were used for this study, yielding 20 paired full depth cartilage samples (half with surface layer intact and half with surface layer removed). Following excision from the bone, samples were allowed to equilibrate first in physiological saline for 2 h, and then in distilled water for another 2 h to maximise the swelling response. Images were captured using a stereomicroscope to measure strain and the fully-swollen samples were fixed in 10% formalin to retain shape for microscopic and ultrastructural imaging. Findings: We expected all swelling samples with an intact 'strain-limiting' surface layer to curl upwards, instead only 70% of them did. For samples without a surface layer, we expected the swelling to be evenly distributed and to remain relatively uncurled; but in 40% of the samples there was a downward curvature (i.e. opposite to that of the previous group). Micro-to-ultrastructural imaging, to determine fibrillar structure and organisation, revealed the deep zone cartilage was an additional counter layer limiting swelling strain, and was the likely cause of the unexpected swelling responses. Interpretation: Our expectations that the surface layer alone will influence the swelling response, was based on the assumptions of the Benninghoff arcade model. This study highlights the additional importance of sub-micron scale fibrillar interconnectivity and the role of the deep zone.