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In vitro expression of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in cultured equine articular cells.
Abstract: To determine concentrations of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in equine chondrocytes and synoviocytes and to quantify changes in the OPG:RANKL ratio in response to exogenous factors. Methods: Samples of articular cartilage and synovium with grossly normal appearance obtained from metacarpophalangeal and metatarsophalangeal joints of 5 adult (1- to 8-year-old) horses. Methods: Cell cultures of chondrocytes and synoviocytes were incubated with human recombinant interleukin-1beta (hrIL-1beta; 10 ng/mL), lipopolysaccharide (LPS; 10 microg/mL), or dexamethasone (100nM) for 48 hours. Negative control cultures received no treatment. Cells and spent media were assayed for RANKL and OPG concentrations by use of western blot and immunocytochemical analyses. Spent media were also assayed for OPG concentration by use of an ELISA. Results: RANKL and OPG were expressed in equine chondrocytes and synoviocytes in vitro. Cell-associated RANKL and OPG concentrations were not impacted by exogenous factors. Soluble RANKL release into media was significantly increased by hrIL-1beta in chondrocyte but not in synoviocyte cultures. Soluble OPG release into media was significantly increased by hrIL-1beta and LPS in chondrocyte but not in synoviocyte cultures. The soluble OPG:RANKL ratio was significantly increased by LPS in chondrocyte cultures. Dexamethasone decreased OPG expression in synoviocytes. Conclusions: RANKL and OPG proteins were expressed in equine articular cells. Release of these proteins may affect osteoclastogenesis within adjacent subchondral bone. Thus, RANKL and OPG may have use as biomarkers and treatment targets in horses with joint disease.
Publication Date: 2010-06-02 PubMed ID: 20513175DOI: 10.2460/ajvr.71.6.615Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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This study explores how articular cells from horse joints produce RANKL and OPG proteins. It also investigates how external factors affect the ratio of OPG to RANKL, and considers the proteins’ potential role in diagnosing and treating joint disease in horses.
Research Context and Methodology
- The objective of the research was to determine the levels of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in horse joint cells (chondrocytes and synoviocytes) and to investigate how these levels change in response to external factors.
- Articular cartilage and synovium samples were obtained from the joints of 5 adult horses. The samples were chosen from joints that appeared normal.
- The cell cultures were then exposed to specific treatments for a period of 48 hours. The treatments were human recombinant interleukin-1beta (hrIL-1beta), lipopolysaccharide (LPS), and dexamethasone. Untreated cultures served as negative controls.
Research Technique and Analysis
- The cellular and media samples were evaluated for RANKL and OPG concentrations via western blot and immunocytochemical analyses. Moreover, the OPG concentration in spent media was also measured using an ELISA (Enzyme-Linked Immunosorbent Assay).
Data and Results
- The results demonstrated that RANKL and OPG were expressed in equine chondrocytes and synoviocytes in vitro.
- It was observed that external elements did not impact the concentrations of cell-associated RANKL and OPG. However, the release of soluble RANKL and OPG into the media showed significant changes in response to the treatments.
- For instance, hrIL-1beta significantly increased the release of soluble RANKL and OPG in chondrocyte cultures, but not in synoviocyte cultures. LPS increased the OPG:RANKL ratio in chondrocyte cultures.
- Dexamethasone was found to reduce OPG expression in synoviocytes.
Conclusions and Implications
- The research concludes that RANKL and OPG proteins are expressed in equine articular cells and their release might influence the formation of osteoclasts within the adjacent subchondral bone. This has implications for understanding and managing joint diseases in horses.
- Consequently, RANKL and OPG can potentially serve as valuable biomarkers and therapeutic targets in horses with joint disease.
Cite This Article
APA
Byron CR, Barger AM, Stewart AA, Pondenis HC, Fan TM.
(2010).
In vitro expression of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in cultured equine articular cells.
Am J Vet Res, 71(6), 615-622.
https://doi.org/10.2460/ajvr.71.6.615 Publication
Researcher Affiliations
- Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801, USA. cbyron@ruffianequine.com
MeSH Terms
- Animals
- Blotting, Western / veterinary
- Cartilage, Articular / cytology
- Cartilage, Articular / metabolism
- Cell Culture Techniques
- Chondrocytes
- Enzyme-Linked Immunosorbent Assay / veterinary
- Horses / metabolism
- Immunohistochemistry / veterinary
- Joints / cytology
- Joints / metabolism
- Osteoprotegerin / biosynthesis
- RANK Ligand / biosynthesis
- Synovial Membrane / cytology
- Synovial Membrane / metabolism
Citations
This article has been cited 6 times.- Sano T, Akeda K, Yamada J, Takegami N, Sudo T, Sudo A. Expression of the RANK/RANKL/OPG system in the human intervertebral disc: implication for the pathogenesis of intervertebral disc degeneration. BMC Musculoskelet Disord 2019 May 17;20(1):225.
- Byron CR, Trahan RA. Comparison of the Effects of Interleukin-1 on Equine Articular Cartilage Explants and Cocultures of Osteochondral and Synovial Explants. Front Vet Sci 2017;4:152.
- Staunton CA, Mobasheri A, Barrett-Jolley R. High hopes for cannabinoid agonists in the treatment of rheumatic diseases. BMC Musculoskelet Disord 2014 Dec 4;15:410.
- Bonnet CS, Williams AS, Gilbert SJ, Harvey AK, Evans BA, Mason DJ. AMPA/kainate glutamate receptors contribute to inflammation, degeneration and pain related behaviour in inflammatory stages of arthritis. Ann Rheum Dis 2015 Jan;74(1):242-51.
- Martínez-Calatrava MJ, Prieto-Potín I, Roman-Blas JA, Tardio L, Largo R, Herrero-Beaumont G. RANKL synthesized by articular chondrocytes contributes to juxta-articular bone loss in chronic arthritis. Arthritis Res Ther 2012 Jun 18;14(3):R149.
- Funck-Brentano T, Lin H, Hay E, Ah Kioon MD, Schiltz C, Hannouche D, Nizard R, Lioté F, Orcel P, de Vernejoul MC, Cohen-Solal ME. Targeting bone alleviates osteoarthritis in osteopenic mice and modulates cartilage catabolism. PLoS One 2012;7(3):e33543.
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