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      Effects of long-term and high-dose administration of glucocorticoids on the cranial cruciate ligament in healthy beagle dogs

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          Abstract

          This study aimed to determine the effects of long-term and high-dose administration of glucocorticoids (GCs) on the histological and mechanical properties of the cranial cruciate ligament (CrCL) in healthy beagle dogs. A synthetic corticosteroid at 2 mg/kg every 12 h was administered for 84 days in nine dogs (18 CrCLs) (GC group). Twenty CrCLs from 12 healthy male beagles were used as the normal control (control group). CrCLs were histologically examined (n = 12 in the GC group and n = 14 in the control group) using hematoxylin-eosin, Alcian-Blue, Elastica-Eosin stains, and immunohistological staining of type 1 collagen and elastin. An additional 12 CrCLs were mechanically tested (n = 6 in the GC and n = 6 in the control groups) to determine failure pattern, maximum tensile strength, maximum stress, elastic modulus, and stress and strain at the transition point. The histological examination revealed a significant increase in interfascicular area and fibrillar disorientation at the tibial attachment in both groups. The ratios of mucopolysaccharide-positive area and positive areas of elastic fibers were significantly higher in the control group than in the GC group. The biomechanical examination demonstrated significantly lower stress at the transition point in the GC group than in the control group. The present study results indicate that high-dose corticosteroids may affect metabolism, such as mucopolysaccharides and elastic fibers production, although the effect on type 1 collagen production is small. These changes of the extracellular matrix had a small effect on the strength of the ligament. This study suggested that the ligamentous changes associated with GC are different from the degeneration observed in spontaneous canine CrCL disease.

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          Most cited references35

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          Where tendons and ligaments meet bone: attachment sites ('entheses') in relation to exercise and/or mechanical load.

          Entheses (insertion sites, osteotendinous junctions, osteoligamentous junctions) are sites of stress concentration at the region where tendons and ligaments attach to bone. Consequently, they are commonly subject to overuse injuries (enthesopathies) that are well documented in a number of sports. In this review, we focus on the structure-function correlations of entheses on both the hard and the soft tissue sides of the junction. Particular attention is paid to mechanical factors that influence form and function and thus to exploring the relationship between entheses and exercise. The molecular parameters indicative of adaptation to mechanical stress are evaluated, and the basis on which entheses are classified is explained. The application of the 'enthesis organ' concept (a collection of tissues adjacent to the enthesis itself, which jointly serve the common function of stress dissipation) to understanding enthesopathies is considered and novel roles of adipose tissue at entheses are reviewed. A distinction is made between different locations of fat at entheses, and possible functions include space-filling and proprioception. The basic anchorage role of entheses is considered in detail and comparisons are explored between entheses and other biological 'anchorage' sites. The ability of entheses for self-repair is emphasized and a range of enthesopathies common in sport are reviewed (e.g. tennis elbow, golfer's elbow, jumper's knee, plantar fasciitis and Achilles insertional tendinopathies). Attention is drawn to the degenerative, rather than inflammatory, nature of most enthesopathies in sport. The biomechanical factors contributing to the development of enthesopathies are reviewed and the importance of considering the muscle-tendon-bone unit as a whole is recognized. Bony spur formation is assessed in relation to other changes at entheses which parallel those in osteoarthritic synovial joints.
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            A General Introduction to Glucocorticoid Biology

            Glucocorticoids (GCs) are steroid hormones widely used for the treatment of inflammation, autoimmune diseases, and cancer. To exert their broad physiological and therapeutic effects, GCs bind to the GC receptor (GR) which belongs to the nuclear receptor superfamily of transcription factors. Despite their success, GCs are hindered by the occurrence of side effects and glucocorticoid resistance (GCR). Increased knowledge on GC and GR biology together with a better understanding of the molecular mechanisms underlying the GC side effects and GCR are necessary for improved GC therapy development. We here provide a general overview on the current insights in GC biology with a focus on GC synthesis, regulation and physiology, role in inflammation inhibition, and on GR function and plasticity. Furthermore, novel and selective therapeutic strategies are proposed based on recently recognized distinct molecular mechanisms of the GR. We will explain the SEDIGRAM concept, which was launched based on our research results.
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              Ligament structure, physiology and function.

              Ligaments are specialized connective tissues with very interesting biomechanical properties. They have the ability to adapt to the complex functions that each are required to perform. While ligaments were once thought to be inert, they are in fact responsive to many local and systemic factors that influence their function within the organism. Injury to a ligament results in a drastic change in its structure and physiology and creates a situation where ligament function is restored by the formation of scar tissue that is biologically and biomechanically inferior to the tissue it replaces. This article will briefly review the basic structure, physiology and function of normal versus healing knee ligaments, referring specifically to what is known about two of the most extensively studied and clinically relevant knee ligaments, the anterior cruciate (ACL) and medial collateral (MCL) ligaments of the knee. Those readers wishing for more comprehensive sources of information on ligament biology and biomechanics are referred to many excellent reviews on these topics.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: InvestigationRole: Visualization
                Role: InvestigationRole: Methodology
                Role: Data curationRole: Formal analysisRole: InvestigationRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: ValidationRole: Writing – review & editing
                Role: SupervisionRole: ValidationRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: MethodologyRole: ResourcesRole: SupervisionRole: ValidationRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                21 January 2022
                2022
                : 17
                : 1
                : e0262206
                Affiliations
                [1 ] Division of Veterinary Surgery, Department of Veterinary Science, Faculty of Veterinary Medicine, Nippon Veterinary and Life Science University, Musashino-shi, Tokyo, Japan
                [2 ] Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America
                University of Lincoln, UNITED KINGDOM
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                https://orcid.org/0000-0002-9233-2496
                https://orcid.org/0000-0002-1207-8644
                Article
                PONE-D-21-25951
                10.1371/journal.pone.0262206
                8782538
                35061786
                b91b7da1-c092-4cfd-a26a-a5e368d368ed
                © 2022 Shimada et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 10 August 2021
                : 20 December 2021
                Page count
                Figures: 10, Tables: 1, Pages: 15
                Funding
                Funded by: the Japan Society for the Promotion of Science
                Award ID: 17K08114
                Award Recipient :
                The author (Y.H) was funded by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (Grant number: 17K08114, URL: https://www.jsps.go.jp/english/e-grants/). The creation of the model dogs in this study was supported by the Grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Biological Tissue
                Connective Tissue
                Ligaments
                Medicine and Health Sciences
                Anatomy
                Biological Tissue
                Connective Tissue
                Ligaments
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
                Mammals
                Dogs
                Biology and Life Sciences
                Zoology
                Animals
                Vertebrates
                Amniotes
                Mammals
                Dogs
                Biology and Life Sciences
                Anatomy
                Histology
                Medicine and Health Sciences
                Anatomy
                Histology
                Biology and Life Sciences
                Biomechanics
                Research and Analysis Methods
                Specimen Preparation and Treatment
                Staining
                Group-Specific Staining
                Hematoxylin Staining
                Biology and Life Sciences
                Biochemistry
                Proteins
                Collagens
                Biology and Life Sciences
                Biochemistry
                Proteins
                Elastin
                Physical Sciences
                Materials Science
                Material Properties
                Elasticity
                Custom metadata
                All relevant data are within the manuscript and its Supporting information files.

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