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      Study of the collagen structure in the superficial zone and physiological state of articular cartilage using a 3D confocal imaging technique

      research-article
      1 , , 1 , 2
      Journal of Orthopaedic Surgery and Research
      BioMed Central

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          Abstract

          Introduction

          The collagen structure in the superficial zone of articular cartilage is critical to the tissue's durability. Early osteoarthritis is often characterized with fissures on the articular surface. This is closely related to the disruption of the collagen network. However, the traditional histology can not offer visualization of the collagen structure in articular cartilage because it uses conventional optical microscopy that does not have insufficient imaging resolution to resolve collagen from proteoglycans in hyaline articular cartilage. This study examines the 3D collagen network of articular cartilage scored from 0 to 2 in the scoring system of International Cartilage Repair Society, and aims to develop a 3D histology for assessing early osteoarthritis.

          Methods

          Articular cartilage was visually classified into five physiological groups: normal cartilage, aged cartilage, cartilage with artificial and natural surface disruption, and fibrillated. The 3D collagen matrix of the cartilage was acquired using a 3D imaging technique developed previously. Traditional histology was followed to grade the physiological status of the cartilage in the scoring system of International Cartilage Repair Society.

          Results

          Normal articular cartilage contains interwoven collagen bundles near the articular surface, approximately within the lamina splendens. However, its collagen fibres in the superficial zone orient predominantly in a direction spatially oblique to the articular surface. With age and disruption of the articular surface, the interwoven collagen bundles are gradually disappeared, and obliquely oriented collagen fibres change to align predominantly in a direction spatially perpendicular to the articular surface. Disruption of the articular surface is well related to the disappearance of the interwoven collagen bundles.

          Conclusion

          A 3D histology has been developed to supplement the traditional histology and study the subtle changes in the collagen network in the superficial zone during early physiological alteration of articular cartilage. The fibre confocal imaging technology used in this study has allowed developing confocal arthroscopy for in vivo studying the chondrocytes in different depth of articular cartilage. Therefore, the current study has potential to develop an in vivo 3D histology for diagnosis of early osteoarthritis.

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          Most cited references47

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          Zinc coordination, function, and structure of zinc enzymes and other proteins.

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            Damage to type II collagen in aging and osteoarthritis starts at the articular surface, originates around chondrocytes, and extends into the cartilage with progressive degeneration.

            Enhanced denaturation of type II collagen fibrils in femoral condylar cartilage in osteoarthritis (OA) has recently been quantitated immunochemically (Hollander, A.P., T.F. Heathfield, C. Webber, Y. Iwata, R. Bourne, C. Rorabeck, and A.R. Poole. 1994. J. Clin. Invest. 93:1722-1732). Using the same antibody that only reacts with denatured type II collagen, we investigated with immunoperoxidase histochemistry (results were graded for analysis) the sites of the denaturation (loss of triple helix) of this molecule in human aging (at autopsy, n= 11) and progressively degenerate (by Mankin grade [MG]) OA (at arthroplasty, n= 51) knee condylar cartilages. Up to 41 yr, most aging cartilages (3 of 4) (MG 0-4) showed very little denaturation. In most older cartilages, (4 of 7) (MG 2-4), staining was observed in the superficial and mid zones. This pattern of collagen II denaturation was also seen in all OA specimens with increased staining extending to the deep zone with increasing MG. Collagen II staining correlated directly both with MG and collagen II denaturation measured by immunoassay. Cartilage fibrillation occurred in OA cartilages with increased penetration of the staining for collagen II denaturation into the mid and deep zones and where denaturation was more pronounced by immunoassay. Thus in both aging and OA the first damage to type II collagen occurs in the superficial and upper mid zone (low MG) extending to the lower mid and deep zones with increasing degeneration (increasing MG). Initial damage is always seen around chondrocytes implicating them in the denaturation of type II collagen.
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              Altered cartilage mechanics and histology in knee osteoarthritis: relation to clinical assessment (ICRS Grade).

              Substantial changes in articular cartilage composition and mechanical properties occur during the development of osteoarthritis (OA). While softening in the initial stage is reported and sometimes used as an indicator of early OA, there is a lack of data relating the macroscopic appearance of cartilage to its mechanical and histological properties in all stages of degeneration. Knowledge about the mechanical quality of the tissue is important for diagnostic reasons and the understanding of the development of OA. The cartilage areas of 21 osteoarthritic human cadaver tibia plateaus were classified using the International Cartilage Repair Society (ICRS) system. A material testing device determined the Young's modulus of the cartilage by unconfined compression. Histological analysis used haematoxylin and eosin staining and Safranin-O staining for the evaluation of the Mankin score. A correlation between increasing ICRS Grade and stiffness reduction was found (R2=0.69). Stiffness values were for ICRS Grades 1, 2 and 3: E1=0.50+/-0.14 MPa, E2=0.37+/-0.13 MPa and E3=0.28+/-0.12 MPa, respectively. The histological evaluation confirmed the ICRS classification (R2=0.74). A moderate correlation between Mankin score and cartilage stiffness was observed (R2=0.47). The results indicate a relation between structural, mechanical and histological changes in all stages of the degeneration. With increasing ICRS Grade the cartilage stiffness, which is primarily influenced by the integrity of the extracellular matrix, decreases. Therefore, methods of stiffness determination such as indentation may be used to characterize cartilage in all stages of OA. However, the data suggest that differentiating between healthy cartilage and ICRS Grade 1 may be difficult using mechanical testing alone.
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                Author and article information

                Journal
                J Orthop Surg
                Journal of Orthopaedic Surgery and Research
                BioMed Central
                1749-799X
                2008
                17 July 2008
                : 3
                : 29
                Affiliations
                [1 ]3D imaging laboratory, the School of Mechanical Engineering, The University of Western Australia, Perth, WA, Australia
                [2 ]Orthopaedic Surgery, School of Surgery and Pathology, The University of Western Australia, Perth, WA, Australia
                Article
                1749-799X-3-29
                10.1186/1749-799X-3-29
                2503973
                18637164
                4046cced-df60-47ce-9aef-d6c33e3b9a2c
                Copyright © 2008 Wu et al; licensee BioMed Central Ltd.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 24 August 2007
                : 17 July 2008
                Categories
                Research Article

                Surgery
                Surgery

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