Immunohistochemical analysis of an equine model of synovitis-induced arthritis

P. G. Todhunter, S. A. Kincaid, R. J. Todhunter, J. R. Kammermann, Brian Johnstone, A. N. Baird, R. R. Hanson, J. M. Wright, H. C. Lin, R. C. Purohit

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Abstract

Objectives - To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone. Animals - 2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2). Procedure - Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 μg/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articular cartilage sections and attached synovial membrane from the third carpal bones were examined by immunohistochemical distribution of interleukin 1β, tumor necrosis factor (TNF)-α, TNF receptors (P55, P75) and 3-B-3(-) epitopes, and by localization of proteoglycans (metachromatic staining). Proteoglycan extracts were assessed by metachromatic staining or western blotting and immunohistochemical staining, using anti-3-B-3 antibodies. Results - Enhanced immunoreactivity for the cytokines and receptors was found in inflamed synovial membrane and noncalcified cartilage (group 2a more than 2b). Metachromasia of the noncalcified cartilage was greater in group-1 than in group-2a and group-2b specimens. In group 2a, chondrocyte hypertrophy and enhanced immunoreactivity for 3-B-3(-) epitope in areas of increased cytokine immunoreactivity suggested possible phenotypic change of the chondrocytes in response to synovitis. Immunohistochemical analysis by western blotting of proteoglycan extracts indicated strong 3-B-3(-) epitope immunolocalization in group-2a, weaker staining in group-2b, and barely detectable stain in group-1 specimens, which correlated with in situ immunolocalization. Conclusions - Intra-articular administration of LPS may be used to induce a synovial environment conducive to increased immunoreactivity of interleukin 1β, TNF-α, and its receptors in equine synovial membrane and articular cartilage. These cytokines may be involved in the early phenotypic change of chondrocytes that is believed to occur in osteoarthritis and is characterized in this study by enhanced 3-B-3(-) epitope immunoreactivity and chondrocyte hypertrophy.

Original languageEnglish (US)
Pages (from-to)1080-1093
Number of pages14
JournalAmerican Journal of Veterinary Research
Volume57
Issue number7
StatePublished - Jul 1996
Externally publishedYes

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synovitis
Synovitis
arthritis
Chondrocytes
cartilage
Horses
chondrocytes
Arthritis
Synovial Membrane
Epitopes
Articular Cartilage
Proteoglycans
Joints
epitopes
Carpal Bones
proteoglycans
Staining and Labeling
tumor necrosis factors
Lipopolysaccharides
horses

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Todhunter, P. G., Kincaid, S. A., Todhunter, R. J., Kammermann, J. R., Johnstone, B., Baird, A. N., ... Purohit, R. C. (1996). Immunohistochemical analysis of an equine model of synovitis-induced arthritis. American Journal of Veterinary Research, 57(7), 1080-1093.

Immunohistochemical analysis of an equine model of synovitis-induced arthritis. / Todhunter, P. G.; Kincaid, S. A.; Todhunter, R. J.; Kammermann, J. R.; Johnstone, Brian; Baird, A. N.; Hanson, R. R.; Wright, J. M.; Lin, H. C.; Purohit, R. C.

In: American Journal of Veterinary Research, Vol. 57, No. 7, 07.1996, p. 1080-1093.

Research output: Contribution to journalArticle

Todhunter, PG, Kincaid, SA, Todhunter, RJ, Kammermann, JR, Johnstone, B, Baird, AN, Hanson, RR, Wright, JM, Lin, HC & Purohit, RC 1996, 'Immunohistochemical analysis of an equine model of synovitis-induced arthritis', American Journal of Veterinary Research, vol. 57, no. 7, pp. 1080-1093.
Todhunter PG, Kincaid SA, Todhunter RJ, Kammermann JR, Johnstone B, Baird AN et al. Immunohistochemical analysis of an equine model of synovitis-induced arthritis. American Journal of Veterinary Research. 1996 Jul;57(7):1080-1093.
Todhunter, P. G. ; Kincaid, S. A. ; Todhunter, R. J. ; Kammermann, J. R. ; Johnstone, Brian ; Baird, A. N. ; Hanson, R. R. ; Wright, J. M. ; Lin, H. C. ; Purohit, R. C. / Immunohistochemical analysis of an equine model of synovitis-induced arthritis. In: American Journal of Veterinary Research. 1996 ; Vol. 57, No. 7. pp. 1080-1093.
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title = "Immunohistochemical analysis of an equine model of synovitis-induced arthritis",
abstract = "Objectives - To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone. Animals - 2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2). Procedure - Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 μg/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articular cartilage sections and attached synovial membrane from the third carpal bones were examined by immunohistochemical distribution of interleukin 1β, tumor necrosis factor (TNF)-α, TNF receptors (P55, P75) and 3-B-3(-) epitopes, and by localization of proteoglycans (metachromatic staining). Proteoglycan extracts were assessed by metachromatic staining or western blotting and immunohistochemical staining, using anti-3-B-3 antibodies. Results - Enhanced immunoreactivity for the cytokines and receptors was found in inflamed synovial membrane and noncalcified cartilage (group 2a more than 2b). Metachromasia of the noncalcified cartilage was greater in group-1 than in group-2a and group-2b specimens. In group 2a, chondrocyte hypertrophy and enhanced immunoreactivity for 3-B-3(-) epitope in areas of increased cytokine immunoreactivity suggested possible phenotypic change of the chondrocytes in response to synovitis. Immunohistochemical analysis by western blotting of proteoglycan extracts indicated strong 3-B-3(-) epitope immunolocalization in group-2a, weaker staining in group-2b, and barely detectable stain in group-1 specimens, which correlated with in situ immunolocalization. Conclusions - Intra-articular administration of LPS may be used to induce a synovial environment conducive to increased immunoreactivity of interleukin 1β, TNF-α, and its receptors in equine synovial membrane and articular cartilage. These cytokines may be involved in the early phenotypic change of chondrocytes that is believed to occur in osteoarthritis and is characterized in this study by enhanced 3-B-3(-) epitope immunoreactivity and chondrocyte hypertrophy.",
author = "Todhunter, {P. G.} and Kincaid, {S. A.} and Todhunter, {R. J.} and Kammermann, {J. R.} and Brian Johnstone and Baird, {A. N.} and Hanson, {R. R.} and Wright, {J. M.} and Lin, {H. C.} and Purohit, {R. C.}",
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T1 - Immunohistochemical analysis of an equine model of synovitis-induced arthritis

AU - Todhunter, P. G.

AU - Kincaid, S. A.

AU - Todhunter, R. J.

AU - Kammermann, J. R.

AU - Johnstone, Brian

AU - Baird, A. N.

AU - Hanson, R. R.

AU - Wright, J. M.

AU - Lin, H. C.

AU - Purohit, R. C.

PY - 1996/7

Y1 - 1996/7

N2 - Objectives - To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone. Animals - 2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2). Procedure - Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 μg/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articular cartilage sections and attached synovial membrane from the third carpal bones were examined by immunohistochemical distribution of interleukin 1β, tumor necrosis factor (TNF)-α, TNF receptors (P55, P75) and 3-B-3(-) epitopes, and by localization of proteoglycans (metachromatic staining). Proteoglycan extracts were assessed by metachromatic staining or western blotting and immunohistochemical staining, using anti-3-B-3 antibodies. Results - Enhanced immunoreactivity for the cytokines and receptors was found in inflamed synovial membrane and noncalcified cartilage (group 2a more than 2b). Metachromasia of the noncalcified cartilage was greater in group-1 than in group-2a and group-2b specimens. In group 2a, chondrocyte hypertrophy and enhanced immunoreactivity for 3-B-3(-) epitope in areas of increased cytokine immunoreactivity suggested possible phenotypic change of the chondrocytes in response to synovitis. Immunohistochemical analysis by western blotting of proteoglycan extracts indicated strong 3-B-3(-) epitope immunolocalization in group-2a, weaker staining in group-2b, and barely detectable stain in group-1 specimens, which correlated with in situ immunolocalization. Conclusions - Intra-articular administration of LPS may be used to induce a synovial environment conducive to increased immunoreactivity of interleukin 1β, TNF-α, and its receptors in equine synovial membrane and articular cartilage. These cytokines may be involved in the early phenotypic change of chondrocytes that is believed to occur in osteoarthritis and is characterized in this study by enhanced 3-B-3(-) epitope immunoreactivity and chondrocyte hypertrophy.

AB - Objectives - To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone. Animals - 2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2). Procedure - Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 μg/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articular cartilage sections and attached synovial membrane from the third carpal bones were examined by immunohistochemical distribution of interleukin 1β, tumor necrosis factor (TNF)-α, TNF receptors (P55, P75) and 3-B-3(-) epitopes, and by localization of proteoglycans (metachromatic staining). Proteoglycan extracts were assessed by metachromatic staining or western blotting and immunohistochemical staining, using anti-3-B-3 antibodies. Results - Enhanced immunoreactivity for the cytokines and receptors was found in inflamed synovial membrane and noncalcified cartilage (group 2a more than 2b). Metachromasia of the noncalcified cartilage was greater in group-1 than in group-2a and group-2b specimens. In group 2a, chondrocyte hypertrophy and enhanced immunoreactivity for 3-B-3(-) epitope in areas of increased cytokine immunoreactivity suggested possible phenotypic change of the chondrocytes in response to synovitis. Immunohistochemical analysis by western blotting of proteoglycan extracts indicated strong 3-B-3(-) epitope immunolocalization in group-2a, weaker staining in group-2b, and barely detectable stain in group-1 specimens, which correlated with in situ immunolocalization. Conclusions - Intra-articular administration of LPS may be used to induce a synovial environment conducive to increased immunoreactivity of interleukin 1β, TNF-α, and its receptors in equine synovial membrane and articular cartilage. These cytokines may be involved in the early phenotypic change of chondrocytes that is believed to occur in osteoarthritis and is characterized in this study by enhanced 3-B-3(-) epitope immunoreactivity and chondrocyte hypertrophy.

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