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      In situ characterization of nanoscale strains in loaded whole joints via synchrotron X-ray tomography

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          Abstract

          Imaging techniques for quantifying how the hierarchical structure of deforming joints changes are constrained by destructive sample treatments, sample-size restrictions and lengthy scan times. Here, we report the use of fast, low-dose pink-beam synchrotron X-ray tomography combined with mechanical loading at nanometric precision for the in situ imaging, at resolutions lower than 100 nm, of mechanical strain in intact untreated joints under physiologically realistic conditions. We show that, in young, aged, and osteoarthritic mice, hierarchical changes in tissue structure and mechanical behaviour can be simultaneously visualized, and that tissue structure at the cellular level correlates with whole-joint mechanical performance. We also used the tomographic approach to study the co-localization of tissue strains to specific chondrocyte lacunar organizations within intact loaded joints, and for the exploration of the role of calcified-cartilage stiffness on the biomechanics of healthy and pathological joints.

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          Most cited references66

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          Articular cartilage and subchondral bone in the pathogenesis of osteoarthritis.

          The articular surface plays an essential role in load transfer across the joint, and conditions that produce increased load transfer or altered patterns of load distribution accelerate the development of osteoarthritis (OA). Current knowledge segregates the risk factors into two fundamental mechanisms related to the adverse effects of "abnormal" loading on normal cartilage or "normal" loading on abnormal cartilage. Although chondrocytes can modulate their functional state in response to loading, their capacity to repair and modify the surrounding extracellular matrix is limited in comparison to skeletal cells in bone. This differential adaptive capacity underlies the more rapid appearance of detectable skeletal changes, especially after acute injuries that alter joint mechanics. The imbalance in the adaptation of the cartilage and bone disrupts the physiological relationship between these tissues and further contributes to OA pathology. This review focuses on the specific articular cartilage and skeletal features of OA and the putative mechanisms involved in their pathogenesis.
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            Ptychographic X-ray computed tomography at the nanoscale.

            X-ray tomography is an invaluable tool in biomedical imaging. It can deliver the three-dimensional internal structure of entire organisms as well as that of single cells, and even gives access to quantitative information, crucially important both for medical applications and for basic research. Most frequently such information is based on X-ray attenuation. Phase contrast is sometimes used for improved visibility but remains significantly harder to quantify. Here we describe an X-ray computed tomography technique that generates quantitative high-contrast three-dimensional electron density maps from phase contrast information without reverting to assumptions of a weak phase object or negligible absorption. This method uses a ptychographic coherent imaging approach to record tomographic data sets, exploiting both the high penetration power of hard X-rays and the high sensitivity of lensless imaging. As an example, we present images of a bone sample in which structures on the 100 nm length scale such as the osteocyte lacunae and the interconnective canalicular network are clearly resolved. The recovered electron density map provides a contrast high enough to estimate nanoscale bone density variations of less than one per cent. We expect this high-resolution tomography technique to provide invaluable information for both the life and materials sciences.
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              The bone-cartilage unit in osteoarthritis.

              Osteoarthritis (OA) refers to a group of mechanically-induced joint disorders to which both genetic and acquired factors contribute. Current pathophysiological concepts focus on OA as a disease of the whole joint. Within these models, the functional unit formed by the articular cartilage and the subchondral bone seems to be of particular interest. Cartilage and bone receive and dissipate the stress associated with movement and loading, and are therefore continuously challenged biomechanically. Recent data support the view that cartilage and bone can communicate over the calcified tissue barrier; vessels reach out from bone into the cartilage zone, patches of uncalcified cartilage are in contact with bone, and microcracks and fissures further facilitate transfer of molecules. Several molecular signaling pathways such as bone morphogenetic proteins and Wnts are hypothesized to have a role in OA and can activate cellular and molecular processes in both cartilage and bone cells. In addition, intracellular activation of different kinase cascades seems to be involved in the molecular crosstalk between cartilage and bone cells. Further research is required to integrate these different elements into a comprehensive approach that will increase our understanding of the disease processes in OA, and that could lead to the development of specific therapeutics or treatment strategies.
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                Author and article information

                Journal
                101696896
                Nat Biomed Eng
                Nat Biomed Eng
                Nature biomedical engineering
                2157-846X
                12 October 2019
                25 November 2019
                March 2020
                25 May 2020
                : 4
                : 3
                : 343-354
                Affiliations
                [1 ]Research Complex at Harwell, RAL, Didcot, OX11 0FA, UK
                [2 ]3Dmagination Ltd, Fermi Avenue, Harwell Oxford, Didcot, OX11 0QR, UK
                [3 ]School of Applied Sciences, Edinburgh Napier University, Edinburgh, EH11 4BN, UK
                [4 ]College of Engineering, Oregon State University, Corvallis, OR, 97331, USA
                [5 ]Comparative Biomedical Sciences, Royal Veterinary College, Royal College Street, London, NW10TU, UK
                [6 ]Department of Materials, The University of Manchester, Manchester, M13 9PL, UK
                [7 ]Diamond Light Source, Harwell Science & Innovation Campus, Oxfordshire, OX11 0DE, UK
                [8 ]Mechanical Engineering, University College London, London, WC1E 7JE, UK
                Author notes
                Article
                EMS84619
                10.1038/s41551-019-0477-1
                7101244
                31768001
                b5457bac-79db-4cde-8613-94b0ccc71757

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