Cartilage-on-cartilage contact: effect of compressive loading on tissue deformations and structural integrity of bovine articular cartilage  L. Zevenbergen,

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Cartilage-on-cartilage contact: effect of compressive loading on tissue deformations and structural integrity of bovine articular cartilage  L. Zevenbergen, W. Gsell, L. Cai, D.D. Chan, N. Famaey, J. Vander Sloten, U. Himmelreich, C.P. Neu, I. Jonkers  Osteoarthritis and Cartilage  Volume 26, Issue 12, Pages 1699-1709 (December 2018) DOI: 10.1016/j.joca.2018.08.009 Copyright © 2018 The Author(s) Terms and Conditions

Fig. 1 A) Schematic diagram of the loading apparatus and imaging space. B) The loading profile was gated with the protocol for the displacement encoding with stimulated echoes (DENSE) imaging to ensure precise timing; the encoding gradient was applied prior to loading at 0.15 s. After the mixing period (0.15–0.65 s), the displacement images were acquired during the loaded phase (0.65–2 s). C) Quantitative imaging protocol; anatomical scan (RARE), relaxation measures (T1, T2, T1ρ) and displacement encoding with stimulated echoes (DENSE) during compressive loading: ≈500 cycles of pre-loading were applied followed by ≈1200 cycles during DENSE imaging. Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

Fig. 2 Locations of the cartilage sub-regions used for statistical analysis of displacement and strain distributions in both explants (medial – top, lateral – bottom of the image). Comparisons were made at three tissue depths; deep zone (DZ), middle zone (MZ) and superficial zone (SZ) and at different transverse locations; two zones without contact (NC1, NC2) and five zones that describe the contacting area (CA1-CA5). Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

Fig. 3 Average cartilage thickness throughout the contact region before and immediately after loading. Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

Fig. 4 Two-dimensional displacement (A), strain (B) and T1, T2 and T1ρ relaxometry data before (C) and after loading (D) within the articular cartilage layers of a representative sample. The displacements and strains were determined by DENSE during cyclic compressive loading along the Y-axis. The top cartilage layer represents the medial osteochondral explant (static) and the opposing layer the lateral explant (indenting). The T1, T2 and T1ρ relaxometry maps were acquired before and immediately after loading using variable repetition time (TR), echo time (TE) and spin-lock durations (TSL), respectively. Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

Fig. 5 Transverse (dX) and axial displacements (dY) at selected regions of interest in the medial (A&D) and lateral cartilage layers (B&E). Displacements were calculated at different tissue depths (DZ = deep zone, MZ = middle zone, SZ = superficial zone) for different transverse locations (NC1, NC2 = non-contacting areas, CA1 – CA5 = contact zone) (Fig. 3). Data are expressed as mean ± standard deviations, n = 8. Pane C and F represent the linear mixed effect model estimates describing the spatial distribution of the transverse and axial displacements. The horizontal lines represent the 95% confidence intervals for the main parameter estimates, the corresponding P-values are given on the right vertical axis. Each model included tissue depth (DZ, MZ, SZ), transverse location (NC1, CA1-CA5, NC2) and explant (med-lat) as fixed effects and subjects as a random effect. The medial explant, central contact region (CA3) and DZ served as reference categories. A negative estimate value indicates that the compared parameter is lower than the reference parameter. The interaction effects and intercept for the described models are presented in supplementary material figure S1. Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

Fig. 6 Green–Lagrange strains; transverse (Exx), axial (Eyy) and shear (Exy) at selected regions of interest in the medial (A, D&G) and lateral cartilage layers (B, E&H). Strains were calculated at different tissue depths (DZ = deep zone, MZ = middle zone, SZ = superficial zone) for different transverse locations (NC1, NC2 = non-contacting areas, CA1 – CA5 = contact zone) (Fig. 3). Data are expressed as mean ± standard deviations, n = 8. Pane C, F and I represent the linear mixed effect model estimates describing the spatial distribution of the transverse, axial and shear strains. The horizontal lines represent the 95% confidence intervals for the main parameter estimates, the corresponding P-values are given on the right vertical axis. Each model included tissue depth (DZ, MZ, SZ), transverse location (NC1, CA1-CA5, NC2) and explant (med-lat) as fixed effects and subjects as a random effect. The medial explant, central contact region (CA3) and DZ served as reference categories. A negative estimate value indicates that the compared parameter is lower than the reference parameter. The interaction effects and intercept for the described models are presented in supplementary material figure S2. Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

Fig. 7 Difference in cartilage T1, T2 and T1ρ relaxation times (immediately after – before loading) at selected regions of interest in the medial (A, D&G) and lateral explant (B, E&H). Relaxation parameters were calculated at different tissue depths (DZ = deep zone, MZ = middle zone, SZ = superficial zone) for different transverse locations (NC1, NC2 = non-contacting areas, CA1 – CA5 = contact zone) (Fig. 3). Data are expressed as mean ± standard deviations, n = 8. Pane C, F and I represent linear mixed effect model estimates describing the spatial distribution the relaxation time difference (ΔT1, ΔT2 & ΔT1ρ). The horizontal lines represent the 95% confidence intervals for the main parameter estimates, the corresponding P-values are given on the right vertical axis. Each model included tissue depth (DZ, MZ, SZ), transverse location (NC1, CA1-CA5, NC2) and explant (med-lat) as fixed effects and subjects as a random effect. The medial explant, central contact region (CA3) and DZ served as reference categories. A negative estimate value indicates that the compared parameter is lower than the reference parameter. The interaction effects and intercept for the described models are presented in supplementary material fig. S4. Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

figs1 Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

figs2 Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

figs3 Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions

figs4 Osteoarthritis and Cartilage 2018 26, 1699-1709DOI: (10.1016/j.joca.2018.08.009) Copyright © 2018 The Author(s) Terms and Conditions