Effects of triamcinolone acetonide, sodium hyaluronate, amikacin sulfate, and mepivacaine hydrochloride, alone and in combination, on morphology and matrix composition of lipopolysaccharide-challenged and unchallenged equine articular cartilage explants.
Abstract: To evaluate the effects of triamcinolone acetonide (TA), sodium hyaluronate (HA), amikacin sulfate (AS), and mepivacaine hydrochloride (MC) on articular cartilage morphology and matrix composition in lipopolysaccharide (LPS)-challenged and unchallenged equine articular cartilage explants. Methods: 96 articular cartilage explants from 4 femoropatellar joints of 2 adult horses. Methods: Articular cartilage explants were challenged with LPS (100 ng/mL) or unchallenged for 48 hours, then treated with TA, HA, AS, and MC alone or in combination for 96 hours or left untreated. Cartilage extracts were analyzed for glycosaminoglycan (GAG) content by dimethyl-methylene blue assay (ng/mg of dry wt). Histomorphometric quantification of total lacunae, empty lacunae, and lacunae with pyknotic nuclei was recorded for superficial, middle, and deep cartilage zones. Results: LPS induced a significant increase in pyknotic nuclei and empty lacunae. Treatment with TA or HA significantly decreased empty lacunae (TA and HA), compared with groups without TA or HA, and significantly decreased empty lacunae of LPS-challenged explants, compared with untreated explants. Treatment with AS or MC significantly increased empty lacunae in unchallenged explants, and these effects were attenuated by TA. Treatment with MC significantly increased empty lacunae and pyknotic nuclei and, in combination with LPS, could not be attenuated by TA. Content of GAG did not differ between unchallenged and LPS-challenged explants or among treatments. Conclusions: Treatment with TA or HA supported chondrocyte morphology in culture and protected chondrocytes from toxic effects exerted by LPS, AS, and MC.
Publication Date: 2008-07-03 PubMed ID: 18593234DOI: 10.2460/ajvr.69.7.861Google Scholar: Lookup
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Summary
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This is a study investigating how triamcinolone acetonide (TA), sodium hyaluronate (HA), amikacin sulfate (AS), and mepivacaine hydrochloride (MC) affect the structure and substance of horse articular cartilage, especially when the cartilage is affected by lipopolysaccharide (LPS). The goal is to understand the impact of these substances on cartilage health in hopes of developing more effective ways to support or repair it.
Research Methodology
- The study made use of 96 articular cartilage explants which were harvested from the femoropatellar joints of two adult horses.
- The explants were divided into two groups, one that was exposed to lipopolysaccharide (LPS), a substance that compromises the health of the cartilage, and another that was not (unchallenged group).
- Both groups were then subjected to treatments of triamcinolone acetonide (TA), sodium hyaluronate (HA), amikacin sulfate (AS), and mepivacaine hydrochloride (MC) individually or in combination.
- Democratic methyl-methylene blue assay was used to analyze the cartilage for glycosaminoglycan (GAG) content, an important compound in the cartilage matrix.
- Histomorphometric scanning was conducted to evaluate the changes in morphology and density of chondrocytes (lacunae) in three specific cartilage zones.
Research Findings
- The LPS challenging process resulted in an increased count of pyknotic nuclei (a sign of cell death) and empty lacunae (spaces where chondrocytes are missing).
- Treatment with TA or HA significantly reduced the occurrence of empty lacunae in both the LPS-challenged and unchallenged explants, indicating a protective effect on the cartilage.
- However, treatments with AS or MC resulted in more empty lacunae in the unchallenged explants, suggesting that these treatments might have harmful effects on the cartilage. But these adverse effects were reduced when TA was introduced.
- Treatment with MC led to an increase in not only empty lacunae but also pyknotic nuclei. This effect worsened when used in combination with LPS, and TA could not reverse it.
- There was no significant difference in GAG content between LPS-challenged and unchallenged explants, or among different treatments.
Conclusion
- The study concludes that TA or HA treatments can support chondrocyte morphology and protect cartilage against toxic effects from LPS, AS, and MC. This discovery informs potential therapeutic treatments for horses suffering from articular cartilage issues.
Cite This Article
APA
Bolt DM, Ishihara A, Weisbrode SE, Bertone AL.
(2008).
Effects of triamcinolone acetonide, sodium hyaluronate, amikacin sulfate, and mepivacaine hydrochloride, alone and in combination, on morphology and matrix composition of lipopolysaccharide-challenged and unchallenged equine articular cartilage explants.
Am J Vet Res, 69(7), 861-867.
https://doi.org/10.2460/ajvr.69.7.861 Publication
Researcher Affiliations
- Comparative Orthopaedic Research Laboratories, Department of Veterinary Clinical Sciences, The Ohio State University, Columbus, OH 43210, USA.
MeSH Terms
- Amikacin / pharmacology
- Animals
- Cartilage, Articular / drug effects
- Cartilage, Articular / metabolism
- Glycosaminoglycans / metabolism
- Histocytochemistry / veterinary
- Horses / metabolism
- Hyaluronic Acid / pharmacology
- Lipopolysaccharides / pharmacology
- Mepivacaine / pharmacology
- Tissue Culture Techniques / veterinary
- Triamcinolone Acetonide / pharmacology
Citations
This article has been cited 12 times.- Pezzanite L, Chow L, Hendrickson D, Gustafson DL, Russell Moore A, Stoneback J, Griffenhagen GM, Piquini G, Phillips J, Lunghofer P, Dow S, Goodrich LR. Evaluation of Intra-Articular Amikacin Administration in an Equine Non-inflammatory Joint Model to Identify Effective Bactericidal Concentrations While Minimizing Cytotoxicity. Front Vet Sci 2021;8:676774.
- Pezzanite L, Chow L, Piquini G, Griffenhagen G, Ramirez D, Dow S, Goodrich L. Use of in vitro assays to identify antibiotics that are cytotoxic to normal equine chondrocytes and synovial cells. Equine Vet J 2021 May;53(3):579-589.
- Kohli S, Tandra V, Gulihar A. Effect of various factors on articular cartilage and their implications on arthroscopic procedures: A review of literature. J Clin Orthop Trauma 2020 May;11(Suppl 3):S396-S401.
- Neuenschwander HM, Moreira JJ, Vendruscolo CP, Fülber J, Seidel SRT, Michelacci YM, Baccarin RYA. Hyaluronic acid has chondroprotective and joint-preserving effects on LPS-induced synovitis in horses. J Vet Sci 2019 Nov;20(6):e67.
- Gilbertie JM, Schnabel LV, Hickok NJ, Jacob ME, Conlon BP, Shapiro IM, Parvizi J, Schaer TP. Equine or porcine synovial fluid as a novel ex vivo model for the study of bacterial free-floating biofilms that form in human joint infections. PLoS One 2019;14(8):e0221012.
- Rubio-Martínez LM, Rioja E, Castro Martins M, Wipawee S, Clegg P, Peffers MJ. Local anaesthetics or their combination with morphine and/or magnesium sulphate are toxic for equine chondrocytes and synoviocytes in vitro. BMC Vet Res 2017 Nov 7;13(1):318.
- Carmona JU, Ríos DL, López C, Álvarez ME, Pérez JE. Proinflammatory and Anabolic Gene Expression Effects of Platelet-Rich Gel Supernatants on Equine Synovial Membrane Explants Challenged with Lipopolysaccharide. Vet Med Int 2017;2017:6059485.
- Carmona JU, Ríos DL, López C, Álvarez ME, Pérez JE, Bohórquez ME. In vitro effects of platelet-rich gel supernatants on histology and chondrocyte apoptosis scores, hyaluronan release and gene expression of equine cartilage explants challenged with lipopolysaccharide. BMC Vet Res 2016 Jul 1;12(1):135.
- Gulihar A, Robati S, Twaij H, Salih A, Taylor GJ. Articular cartilage and local anaesthetic: A systematic review of the current literature. J Orthop 2015 Dec;12(Suppl 2):S200-10.
- Syed HM, Green L, Bianski B, Jobe CM, Wongworawat MD. Bupivacaine and triamcinolone may be toxic to human chondrocytes: a pilot study. Clin Orthop Relat Res 2011 Oct;469(10):2941-7.
- Linde P, Chow L, Sabino I, Williams Z, Impastato R, Dow S, Pezzanite L. Innate immune pathway activation to modulate mesenchymal stromal cell (MSC) interactions with synovium and cartilage. Front Bioeng Biotechnol 2025;13:1605148.
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