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. 2020 Sep 15;10:15106. doi: 10.1038/s41598-020-72208-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Details of image analysis, post processing, and histological referencing. (a) Mid-sagittal Proton Density-weighted images of a representative cartilage sample subject to increasing loading intensity, i.e. at the unloaded configuration (δ₀) and under loading to 15.1 N (δ₁), and 28.6 N (δ₂). (b) Corresponding T2 maps overlaid onto the respective morphological images (TE 41.9 ms). Unit of scale on the right is [ms]. (c) Regional and zonal ROIs were defined as the upper and lower sample halves as well as the sub-pistonal (SPA) and peri-pistonal area (PPA). Consequently, seven ROIs were evaluated: (1) SPA_upper, (2) SPA_lower, (3) PPA_upper, (4) PPA_lower, (5) SPA full thickness (1 + 2), (6) PPA full thickness (3 + 4), (7) entire cartilage sample (ECS = 1 + 2 + 3 + 4). (d) Corresponding histological sections, stained with hematoxylin/eosin (d₁,d₃) and Safranin O (d₂,d₄), of the cartilage sample following incubation without enzyme exposure and pre- and post-exposure measurements (d₁,d₂) and of the adjacent cartilage tissue at the time of sample preparation (d₃,d₄). Same cartilage sample as in Fig. 4 (control).