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Home / Equine Research Bank / Vet Surg / Clodronate disodium is neither cytotoxic nor cytoprotective ...
Veterinary surgery : VS2022; 52(1); 146-156; doi: 10.1111/vsu.13898

Clodronate disodium is neither cytotoxic nor cytoprotective to normal and recombinant equine interleukin-1β-treated joint tissues in vitro.

Abstract: To determine the effects of clodronate disodium (CLO) on control and recombinant equine interleukin-1β (IL-1β)-treated equine joint tissues. Methods: In vitro experimental study. Methods: Cartilage explants, chondrocytes, and synoviocytes (n = 3 horses). Methods: Monolayer cultures of chondrocytes and synoviocytes from three horses were subjected to: control media (CON), 5 ng/ml CLO (C/low), 50 ng/ml CLO (C/med), 100 ng/ml CLO (C/high), with and without IL-1β, and 10 ng/ml IL-1β (IL) alone for 72 hours. Cartilage explants from three horses were subjected to CON, IL, C/low, and C/med with and without IL-1β for 72 hours. Culture media was analyzed for prostaglandin-E2 (PGE2 ), interleukin-6 (IL-6), and nitric oxide (NO). Explant media was analyzed for glycosaminoglycan (GAG) content and NO. At 72 hours, explant and monolayer culture viability were assessed, and explant GAG content was measured. Results: IL-1β treatment resulted in higher media concentrations of GAG, NO, PGE2 , and IL-6 compared to the CON treatment (p < .05), demonstrating a catabolic effect of IL-1β on explants and monolayer cultures. CLO treatments did not increase media concentrations of GAG, NO, PGE2 , or IL-6 compared to CON, indicating no cytotoxic effect. Nevertheless, CLO treatments administered to IL-1β-treated monolayer cultures and explants did not significantly reduce the inflammatory response regardless of concentration. Conclusions: CLO did not demonstrate cytotoxic nor cytoprotective effects in normal and IL-1β-stimulated chondrocytes, synoviocytes or explants in culture. Conclusions: This study does not support the use of CLO as an anti-inflammatory treatment. Further research is necessary to confirm any anti-inflammatory effects of CLO on joint tissues.
© 2022 American College of Veterinary Surgeons.
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Publication Date: 2022-10-10 PubMed ID: 36217704DOI: 10.1111/vsu.13898Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research paper investigates the impact of clodronate disodium (CLO) on both normal and interleukin-1β (IL-1β)-treated equine joint tissues, concluding that the substance neither harms nor protects the tissues and thus should not be used as an anti-inflammatory treatment.

Research Methods

  • The researchers undertook an in vitro experimental study involving monolayer cultures of chondrocytes and synoviocytes (the cellular components of cartilage and the synovial membrane in joints respectively) derived from three horses.
  • These tissues were subjected to various conditions: a control environment, CLO concentrations at three different levels, with and without IL-1β present, and IL-1β alone, for a duration of 72 hours.
  • Additionally, cartilage explants (pieces of tissue extracted to be studied in a different environment) from the same horses were exposed to similar conditions.
  • The researchers then analyzed the culture media (the substance in which the cells were grown) for concentrations of prostaglandin-E (PGE), interleukin-6 (IL-6), and nitric oxide (NO), which are all markers of inflammation and cell damage.
  • For the explant media, they also counted the amount of glycosaminoglycan (GAG), a compound that forms a vital component of the joint’s cartilage.

Research Findings

  • It was found that IL-1β treatment led to greater media concentrations of GAG, NO, PGE, and IL-6 than in controls, indicating that IL-1β has a destructive effect on explants and monolayer cultures.
  • In contrast, CLO treatments did not increase the media amounts of these compounds, suggesting no harmful effect from CLO.
  • However, administering CLO to IL-1β-treated cultures and explants did not significantly reduce the inflammatory response, regardless of the CLO concentration. These findings demonstrate that CLO lacks a protective effect on these cultures and explants.

Conclusions

  • The study concludes that clodronate disodium does not demonstrate either cytotoxic (harmful to cells) or cytoprotective (protective of cells) effects in both normal and IL-1β-stimulated chondrocytes, synoviocytes, or explants in culture.
  • Subsequently, they do not support the use of CLO as an anti-inflammatory treatment. They suggest further research to confirm any potential anti-inflammatory effects of CLO on joint tissues.

Cite This Article

APA
Vergara-Hernandez FB, Panek CL, Nielsen BD, Robison CI, Colbath AC. (2022). Clodronate disodium is neither cytotoxic nor cytoprotective to normal and recombinant equine interleukin-1β-treated joint tissues in vitro. Vet Surg, 52(1), 146-156. https://doi.org/10.1111/vsu.13898

Publication

Veterinary surgery : VS
ISSN: 1532-950X
NlmUniqueID: 8113214
Country: United States
Language: English
Volume: 52
Issue: 1
Pages: 146-156

Researcher Affiliations

Vergara-Hernandez, Fernando B
  • Department of Animal Science, College of Agricultural and Natural Resources, Michigan State University, East Lansing, Michigan, USA.
Panek, Char L
  • Department of Large Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan, USA.
Nielsen, Brian D
  • Department of Animal Science, College of Agricultural and Natural Resources, Michigan State University, East Lansing, Michigan, USA.
Robison, Cara I
  • Department of Animal Science, College of Agricultural and Natural Resources, Michigan State University, East Lansing, Michigan, USA.
Colbath, Aimee C
  • Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.

MeSH Terms

  • Animals
  • Horses
  • Interleukin-1beta / pharmacology
  • Interleukin-1beta / metabolism
  • Clodronic Acid / pharmacology
  • Clodronic Acid / metabolism
  • Cartilage, Articular
  • Interleukin-6 / metabolism
  • Interleukin-6 / pharmacology
  • Chondrocytes
  • Glycosaminoglycans / pharmacology
  • Glycosaminoglycans / metabolism
  • Antineoplastic Agents / pharmacology

Grant Funding

  • American College of Veterinary Surgeons Foundation

References

This article includes 39 references
  1. Russell RGG. Bisphosphonates: the first 40 years. Bone 2011;49(1):2-19.
  2. Mitchell A, Watts AE, Ebetino FH, Suva LJ. Bisphosphonate use in the horse: what is good and what is not?. BMC Vet Res 2019;15(1):1-7.
    doi: 10.1186/s12917-019-1966-xgoogle scholar: lookup
  3. Anastasilakis AD, Polyzos SA, Makras P. Review Denosumab vs bisphosphonates for the treatment of postmenopausal osteoporosis. Eur J Endocrinol 2018;179(1):31-45.
  4. Baykan EK, Saygili LF, Erdogan M, Cetinkalp S, Ozgen AG, Yilmaz C. Efficacy of zoledronic acid treatment in Paget disease of bone. Osteoporos Int 2014;25(9):2221-2223.
  5. D'Eufemia P, Finocchiaro R, Celli M. High levels of serum prostaglandin E2 in children with osteogenesis imperfecta are reduced by neridronate treatment. Pediatr Res 2008;63(2):203-206.
  6. U.S. Food and Drug Administration. Animal Drugs @ FDA. Accessed June 20, 2022. https://animaldrugsatfda.fda.gov/adafda/views/#/home/previewsearch/141-420
  7. Maksymowych WP. Bisphosphonates-anti-inflammatory properties. Antiinflamm Antiallergy Agents Med Chem 2002;1:15-28.
  8. Garganta MD, Jaser SS, Lazow MA. Cyclic bisphosphonate therapy reduces pain and improves physical functioning in children with osteogenesis imperfecta. BMC Musculoskelet Disord 2018;19(1):344.
    doi: 10.1186/s12891-018-2252-ygoogle scholar: lookup
  9. Mitchell A, Wright G, Sampson SN. Clodronate improves lameness in horses without changing bone turnover markers. Equine Vet J 2019;51(3):356-363.
  10. Duesterdieck-Zellmer KF, Moneta L, Ott JF. Concentration-dependent effects of tiludronate on equine articular cartilage explants incubated with and without interleukin-1β. Am J Vet Res 2012;73(10):1530-1539.
  11. Mönkkönen J, Pennanen N, Lapinjoki S, Urtti A. Clodronate (dichloromethylene bisphosphonate) inhibits LPS-stimulated IL-6 and TNF production by RAW 264 cells. Life Sci 1994;54(14):PL229-PL234.
  12. Denoix JM, Thibaud D, Riccio B. Tiludronate as a new therapeutic agent in the treatment of navicular disease: a double-blind placebo-controlled clinical trial. Equine Vet J 2003;35(4):407-413.
  13. Coudry V, Thibaud D, Riccio B, Audigié F, Didierlaurent D, Denoix JM. Efficacy of tiludronate in the treatment of horses with signs of pain associated with osteoarthritic lesions of the thoracolumbar vertebral column. Am J Vet Res 2007;68(3):329-337.
  14. Gough MR, Thibaud D, Smith RKW. Tiludronate infusion in the treatment of bone spavin: a double blind placebo-controlled trial. Equine Vet J 2010;42(5):381-387.
  15. Bertuglia A, Basano I, Pagliara E, Bottegaro NB, Spinella G, Bullone M. Effect of intravenous tiludronate disodium administration on the radiographic progression of osteoarthritis of the fetlock joint in Standardbred racehorses. J Am Vet Med Assoc 2021;259(6):651-661.
  16. Krueger CR, Mitchell CF, Leise BS, Knych HK. Pharmacokinetics and pharmacodynamics of clodronate disodium evaluated in plasma, synovial fluid and urine. Equine Vet J 2020;52(5):725-732.
  17. Van Offel JF, Schuerwegh AJ, Bridts CH, Stevens WJ, De LS. Effect of bisphosphonates on viability, proliferation, and dexamethasone-induced apoptosis of articular chondrocytes. Ann Rheum Dis 2002;61:925-928.
  18. Barrera P, Blom A, Van Lent PL. Synovial macrophage depletion with clodronate-containing liposomes in rheumatoid arthritis. Arthritis Rheum 2000;43(9):1951-1959.
  19. Polfliet MM, Goede PH, van Kesteren-Hendrikx EM, van Rooijen N, Dijkstra CD, van den Berg TK. A method for the selective depletion of perivascular and meningeal macrophages in the central nervous system. J Neuroimmunol 2001;116(2):188-195.
  20. McIlwraith CW, Frisbie DD, Kawcak CE. The horse as a model of naturally occurring osteoarthritis. Bone Joint Res 2012;1(11):297-309.
  21. Hussein MR, Fathi NA, Ezz El-Din AM. Alterations of the CD4+, CD8+ T cell subsets, interleukins-1β, IL-10, IL-17, tumor necrosis factor-α and soluble intercellular adhesion molecule-1 in rheumatoid arthritis and osteoarthritis: preliminary observations. Pathol Oncol Res 2008;14:321-328.
  22. Noss EH, Brenner MB. The role and therapeutic implications of fibroblast-like synoviocytes in inflammation and cartilage erosion in rheumatoid arthritis. Immunol Rev 2008;223:252-270.
  23. Takafuji VA, McIlwraith CW, Howard RD. Effects of equine recombinant interleukin-1α and interleukin-1β on proteoglycan metabolism and prostaglandin E2 synthesis in equine articular cartilage explants. Am J Vet Res 2002;63(4):551-558.
  24. Dechant JE, Baxter GM, Frisbie DD, Trotter GW, McIlwraith CW. Effects of dosage titration of methylprednisolone acetate and triamcinolone acetonide on interleukin-1-conditioned equine articular cartilage explants in vitro. Equine Vet J 2003;35(5):444-450.
  25. Fenton JI, Chlebek-Brown KA, Caron JP, Orth MW. Effect of glucosamine on interleukin-1-conditioned articular cartilage. Equine Vet J 2002;34(S34):219-223.
  26. Nakano K, Boyle DL, Firestein GS. Regulation of DNA methylation in rheumatoid arthritis synoviocytes. J Immunol 2013;190:1297-1303.
  27. Dombrecht EJ, Schuerwegh AJ, Bridts CH. Effect of bisphosphonates on nitric oxide production by inflammatory activated chondrocytes. Clin Exp Rheumatol 2007;25(6):817-822.
  28. Van Offel JF, Dombrecht EJ, Bridts CH. Influence of bisphosphonates on the production of pro-inflammatory cytokines by activated human articular chondrocytes. Cytokine 2005;31(4):298-304.
  29. Blanco FJ, Ochs RL, Schwarz H, Lotz M. Chondrocyte apoptosis induced by nitric oxide. Am J Pathol 1995;146(1):75-85.
  30. Kumar P, Nagarajan A, Uchil PD. Analysis of cell viability by the MTT assay. Cold Spring Harb Protoc 2018;6:469-472.
  31. Chandrasekhar S, Esterman MA, Hoffman HA. Microdetermination of proteoglycans and glycosaminoglycans in the presence of guanidine hydrochloride. Anal Biochem 1987;161(1):103-108.
  32. Soto SA, Barbará AC. Bisphosphonates: pharmacology and clinical approach to their use in equine osteoarticular diseases. J Equine Vet Sci 2014;34(6):727-737.
  33. van Weeren PR, de Grauw JC. Pain in osteoarthritis. Vet Clin North Am Equine Pract 2010;26(3):619-642.
  34. Chen Y, Jiang W, Yong H. Macrophages in osteoarthritis: pathophysiology and therapeutics. Am J Transl Res 2020;12(1):261-268.
  35. Ewers BJ, Dvoracek-Driksna D, Orth MW, Haut RC. The extent of matrix damage and chondrocyte death in mechanically traumatized articular cartilage explants depends on rate of loading. J Orthop Res 2001;19(5):779-784.
  36. Fenton JI, Chlebek-Brown KA, Peters TL, Caron JP, Orth MW. Glucosamine HCl reduces equine articular cartilage degradation in explant culture. Osteoarthr Cartil 2000;8(4):258-265.
  37. Makkonen N, Hirvonen MR, Teräväinen T, Savolainen K, Mönkkönen J. Different effects of three bisphosphonates on nitric oxide production by RAW 264 macrophage-like cells in vitro. J Pharmacol Exp Ther 1996;277(2):1097-1102.
  38. Moriyama Y, Nomura M. Clodronate: a vesicular ATP release blocker. Trends Pharmacol Sci 2018;39(1):13-23.
  39. Zellmer S, Schmidt-Heck W, Godoy P. Transcription factors ETF, E2F, and SP-1 are involved in cytokine-independent proliferation of murine hepatocytes. Hepatology 2010;52(6):2127-2136.
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