Immunohistochemical analysis of structural changes in collagen for the assessment of osteoarthritis

Tian, Y., Peng, Z., Gorton, D., Xiao, Y., and Ketheesan, N. (2011) Immunohistochemical analysis of structural changes in collagen for the assessment of osteoarthritis. Proceedings of the Institution of Mechanical Engineers, Part H: Journal of Engineering in Medicine, 225 (7). pp. 680-687.

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Collagen fibrillation within articular cartilage (AC) plays a key role in joint osteoarthritis (OA) progression and, therefore, studying collagen synthesis changes could be an indicator for use in the assessment of OA. Various staining techniques have been developed and used to determine the collagen network transformation under microscopy. However, because collagen and proteoglycan coexist and have the same index of refraction, conventional methods for specific visualization of collagen tissue is difficult. This study aimed to develop an advanced staining technique to distinguish collagen from proteoglycan and to determine its evolution in relation to OA progression using optical and laser scanning confocal microscopy (LSCM). A number of AC samples were obtained from sheep joints, including both healthy and abnormal joints with OA grades 1 to 3. The samples were stained using two different trichrome methods and immunohistochemistry (IHC) to stain both colourimetrically and with fluorescence. Using optical microscopy and LSCM, the present authors demonstrated that the IHC technique stains collagens only, allowing the collagen network to be separated and directly investigated. Fluorescently-stained IHC samples were also subjected to LSCM to obtain three-dimensional images of the collagen fibres. Changes in the collagen fibres were then correlated with the grade of OA in tissue. This study is the first to successfully utilize the IHC staining technique in conjunction with laser scanning confocal microscopy. This is a valuable tool for assessing changes to articular cartilage in OA.

Item ID: 20834
Item Type: Article (Research - C1)
ISSN: 2041-3033
Keywords: collagen structure, trichrome staining, immunohistochemical staining technique, optical and laser scanning confocal microscopy, osteoarthritis, articular cartilage
Date Deposited: 29 Mar 2012 05:44
FoR Codes: 09 ENGINEERING > 0903 Biomedical Engineering > 090399 Biomedical Engineering not elsewhere classified @ 50%
11 MEDICAL AND HEALTH SCIENCES > 1107 Immunology > 110799 Immunology not elsewhere classified @ 50%
SEO Codes: 92 HEALTH > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920116 Skeletal System and Disorders (incl. Arthritis) @ 100%
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