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• W3019253188 abstract "Purpose: Biomechanical behavior of the human meniscus depends on tissue internal structure and composition, which are known to change in osteoarthritis (OA). Specifically, OA increases proteoglycan biosynthesis that, in turn, increases both osmotic pressure and swelling causing changes in the organization of collagenous extracellular matrix. The aims of this study were: 1) to analyze microstructural organization of human meniscus ex vivo samples in 3D using micro-computed tomography (μCT), and 2) to compare local microstructural organization between menisci from end-stage OA patients and healthy donors. We hypothesize that the collagen network disorganization increases in osteoarthritic meniscus and that main orientation shifts towards the degrading surface. Methods: Human meniscus samples from the Lund musculoskeletal MENIX biobank were retrieved for this study. Both the medial and lateral posterior horns of human menisci from 10 (5 male, 5 female) end-stage medial compartment knee OA patients undergoing total knee replacement (TKR) were used to represent osteoarthritic tissue. As a reference, 10 medial and lateral menisci from deceased adult donors (5 male, 5 female) without a known history of knee OA and without apparent OA change were used. A few millimeter thick tissue section was cut from the posterior horn for μCT analyses. Samples were dehydrated with an ascending ethanol series and treated with hexamethyldisilazane (HMDS) before air-drying them at room temperature. For μCT, we used a desktop device (SkyScan 1272, Bruker microCT) with the following settings: source voltage = 40kV, current = 250μA, pixel size = 2.0 μm, exposure = 1815ms, resolution = 4032x2688, and no filter. Acquired projections were reconstructed using NRecon software (Bruker microCT, V 1.1.7). A size of 1800μm x 1800μm x 1800μm volume of interest (VOI) was selected from the middle of the posterior horn piece (Figure 1). Local orientation analysis in 3D was done by calculating the greyscale gradients for each voxel by comparing greyscale values within a 9 voxel radius. Smallest gradient change was chosen as the orientation direction (angle) of the voxel. Mean, median, quartiles, skewness and kurtosis were calculated as descriptive statistics from the local orientation angle histograms. Group-wise comparison of descriptive parameters were conducted with a Linear Mixed Model in SPSS (IBM SPSS Statistics V.25) with group as factor and a random intercept to account for depence of two menisci from one knee. Results: In HMDS-treated meniscal samples, collagen fiber bundles could be clearly depicted in 3D using μCT. In both medial and lateral horns of donor meniscus, bundles were aligned towards the tip of meniscus. However, in the medial menisci from OA patients, organized bundle structure was lost and the main orientation was shifted towards the degraded surface on the femoral side. Figure 2 shows histograms of orientation angles for each group, and Figure 3 shows calculated parameters (mean values ± standard deviation). Statistical comparison between the groups showed lower orientation angles in medial OA group compared to all other groups (20.8 to 22.7 degrees; Figure 3). The orientation in the lateral OA group was similar to healthy lateral group (mean difference = 0.38 degrees [95% CI = -11.1-11.1]). Conclusions: HDMS-based μCT-imaging method allows visualization and quantitative measurement of the collagen fiber bundles in 3D in the human meniscus ex vivo. In OA, the medial meniscus compartment showed an increase in collagen disorganization with an overall lower orientation angle of remaining bundles being towards the degenerated surface. This could be expected since enrolled patients had unicompartemental end-stage medial OA. To conclude, quantitative analysis of organization of microstructure of human meniscus ex vivo in 3D is feasible with our μCT-based approach.View Large Image Figure ViewerDownload Hi-res image Download (PPT)View Large Image Figure ViewerDownload Hi-res image Download (PPT)" @default.
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• W3019253188 date "2020-04-01" @default.
• W3019253188 modified "2023-10-14" @default.
• W3019253188 title "Ex vivo analysis of local orientation of collagen fiber bundles in 3D in posterior horn human meniscus using micro-computed tomography" @default.
• W3019253188 doi "https://doi.org/10.1016/j.joca.2020.02.431" @default.
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