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Journal of animal science2025; 103; skaf393; doi: 10.1093/jas/skaf393

Effects of clodronate disodium on markers of inflammation and cartilage metabolism in juvenile horses challenged with intra-articular lipopolysaccharide.

Abstract: Perceived chondroprotective and anti-inflammatory benefits of bisphosphonates in the juvenile horse has led to extra-label use without supportive data regarding intra-articular effects on cartilage metabolism and inflammation. Thirty-two yearling Quarter Horses were stratified into 4 treatment groups by age (500 ± 13 d), BW (336 ± 26 kg), sex (n = 16 female; n = 16 male) and initial bone optical density for a 140-d study. The study consisted of two exercise phases: Phase 1 (d 0-84) emulated sales preparation and Phase 2 (d 99-140) mimicked early exercise training. Horses were housed individually (3.6 m × 7.3 m stalls) and fed to meet nutritional requirements. All horses received iso-volumetric intramuscular injections of 1.8 mg/kg BW clodronate disodium (CD) (OSPHOS) or saline (vehicle) on d 0, 42, 84, and 126. Specifically, the treatment groups consisted of the saline control (0×; n = 8), and clodronate injected once (1×; n = 8; d 84), twice (2×; n = 8; d 0, 84), or four times (4×; n = 8; d 0, 42, 84, 126). Horses underwent an intra-articular lipopolysaccharide (LPS) challenge post treatment administration on d 126. Synovial fluid was collected prior to LPS injection (h 0) and at 6, 12, 24, and 336 h post injection. Synovial fluid concentrations of carboxypropeptide of type II collagen (CPII), collagenase cleavage neopeptide (C2C), and prostaglandin E2 (PGE2) were determined by ELISA. Intra-articular LPS increased CPII, C2C, PGE2 and CPII: C2C (P ≤ 0.01). There tended to be a treatment × time interaction in CPII (P = 0.06) where CPII was greatest in 2× with 1× and 0× being the lowest, 4× concentrations were between this range at 24 h post-injection. Likewise, CPII: C2C tended to be influenced by treatment (P = 0.06) with 2× and 4× having greater synthesis: degradation ratio than 1× and 0×. There was a treatment × time × carpus interaction (P = 0.02) in which 1×, 2× and 4× CD groups had greater synovial PGE2 concentrations at 6 h post-injection in the LPS joint compared to 0×. The results of this study indicate that administration of CD increases intra-articular inflammation (PGE2) but does not affect cartilage degradation (C2C) and only tends to increase cartilage synthesis (CPII) according to biomarkers measured. Extra-label bisphosphonate use has occurred in the juvenile horse despite a lack of scientific evidence, due to perceived anti-inflammatory and chondroprotective benefits. The purpose of this study was to evaluate the effects of clodronate disodium (CD) on biomarkers of cartilage metabolism and intra-articular inflammation within the articulating joint of the juvenile horse using an intra-articular inflammatory model. Thirty-two yearling Quarter Horses were treated with either saline, 1-dose, 2-doses or 4-doses of CD for a 140-d study. Carpal circumference and synovial fluid were collected prior to and after initiation of the inflammatory model. Synovial fluid samples were analyzed for biomarkers of cartilage synthesis and degradation as well as intra-articular inflammation. There was no difference between treatments concerning the cartilage degradation biomarker used in this study, and CD only tended to increase cartilage synthesis and carpal circumference after two doses. Treatment by CD did not decrease inflammation, but rather increased inflammation following initiation of the inflammatory challenge. At no point during the sampled timepoints did CD decrease inflammation compared to the control. This initial investigation into the effects of CD on the juvenile articulating joint, laying the groundwork for further investigations into the effects of CD administration in the juvenile synovial joint.
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Publication Date: 2025-11-12 PubMed ID: 41223152PubMed Central: PMC12646851DOI: 10.1093/jas/skaf393Google 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.

Overview

  • This study investigated the effects of clodronate disodium (CD), a bisphosphonate, on cartilage metabolism and inflammation markers in juvenile horses, specifically after inducing joint inflammation using lipopolysaccharide (LPS).
  • The research aimed to understand whether CD offers anti-inflammatory or chondroprotective benefits in the joints of young horses, as it is sometimes used extra-label without supporting data.

Study Design and Subjects

  • Participants: 32 young Quarter Horses (~500 days old, 336 kg average weight, evenly split by sex).
  • Duration: 140 days, with two exercise phases mimicking sales preparation and early training.
  • Housing and Feeding: Individually housed in stalls and fed to meet nutritional needs.
  • Treatment Groups:
    • Control group (0×): received saline injections (n=8).
    • 1× CD: received one clodronate injection on day 84 (n=8).
    • 2× CD: received two injections, on days 0 and 84 (n=8).
    • 4× CD: received four injections on days 0, 42, 84, and 126 (n=8).
  • Dosage: 1.8 mg/kg body weight of clodronate disodium intramuscularly.
  • Inflammatory Challenge: Injection of LPS into the joint on day 126 to induce inflammation.

Data Collection and Biomarkers Measured

  • Synovial fluid sampling times: before LPS injection (baseline) and at 6, 12, 24, and 336 hours after LPS challenge.
  • Biomarkers measured by ELISA:
    • CPII (Carboxypropeptide of type II collagen) – marker of cartilage synthesis.
    • C2C (Collagenase cleavage neopeptide) – marker of cartilage degradation.
    • PGE2 (Prostaglandin E2) – marker of intra-articular inflammation.
  • Other measurements included carpal circumference as an indicator of joint swelling/inflammation.

Main Findings

  • Inflammatory Response to LPS:
    • LPS injection significantly increased CPII, C2C, PGE2, and the CPII:C2C ratio, indicating active inflammation, cartilage synthesis, and degradation.
  • Effect of CD on Cartilage Metabolism:
    • Cartilage degradation (C2C) was not significantly affected by CD treatment.
    • There was a tendency for increased cartilage synthesis (CPII) in horses receiving two doses of CD; four doses showed intermediate levels.
    • The CPII:C2C ratio (reflecting balance between synthesis and degradation) tended to be higher in 2× and 4× CD groups compared to 1× and control, suggesting more synthesis relative to degradation.
  • Effect of CD on Inflammation:
    • Contrary to expectations, CD groups (1×, 2×, 4×) had higher synovial PGE2 levels than controls 6 hours after LPS injection, indicating increased inflammation rather than suppression.
    • No time point showed a decrease in inflammation in CD-treated horses versus controls.
  • Carpal circumference, an indicator of swelling, tended to increase after two doses of CD.

Conclusions and Implications

  • This study provides initial scientific evidence regarding intra-articular effects of clodronate disodium in juvenile horses.
  • While CD did not reduce inflammation and appeared to increase inflammatory marker PGE2, it showed a tendency to promote cartilage synthesis without affecting degradation.
  • Findings suggest caution when using CD extra-label in young horses based on assumed anti-inflammatory or chondroprotective effects, as these assumptions were not supported by this data.
  • The research lays groundwork for further studies on bisphosphonate use in juvenile equine joints to better understand safety and efficacy.

Cite This Article

APA
George JM, Leatherwood JL, Paris BL, Arnold CE, Glass KG, Conrad MB, Martinez RE, Vergara-Hernandez FB, Nielsen BD, Colbath AC, Welsh TH, Bradbery AN. (2025). Effects of clodronate disodium on markers of inflammation and cartilage metabolism in juvenile horses challenged with intra-articular lipopolysaccharide. J Anim Sci, 103, skaf393. https://doi.org/10.1093/jas/skaf393

Publication

Journal of animal science
ISSN: 1525-3163
NlmUniqueID: 8003002
Country: United States
Language: English
Volume: 103
PII: skaf393

Researcher Affiliations

George, James M
  • Department of Animal Science, Texas A&M University, College Station, TX 77843.
  • Department of Animal Science, Tarleton State University, Stephenville, TX 76402.
Leatherwood, Jessica L
  • Department of Animal Science, Texas A&M University, College Station, TX 77843.
  • Department of Animal Science, Tarleton State University, Stephenville, TX 76402.
Paris, Brittany L
  • Department of Animal Science, Texas A&M University, College Station, TX 77843.
  • School of Agricultural Sciences, Sam Houston State University, Huntsville, TX 77341.
Arnold, Carolyn E
  • Department of Large Animal Clinical Sciences, Texas A&M University, College Station, TX 77843.
  • School of Veterinary Medicine, Texas Tech University, Amarillo, TX 79106.
Glass, Kati G
  • Department of Large Animal Clinical Sciences, Texas A&M University, College Station, TX 77843.
Conrad, Matthew B
  • Department of Animal and Range Sciences, Montana State University, Bozeman, MT 59717.
Martinez, Rafael E
  • Department of Animal Science, Texas A&M University, College Station, TX 77843.
  • Department of Animal Science, Tarleton State University, Stephenville, TX 76402.
Vergara-Hernandez, Fernando B
  • Escuela de Medicina Veterinaria, Facultad de Recursos Naturales y Medicina Veterinaria, Universidad Santo Tomás, Viña del Mar, 2561780, Chile.
Nielsen, Brian D
  • Department of Animal Science, Michigan State University, East Lansing, MI 48824.
Colbath, Aimee C
  • Department of Clinical Sciences, Cornell University, Ithaca, NY 14853.
Welsh, Thomas H
  • Department of Animal Science, Texas A&M University, College Station, TX 77843.
Bradbery, Amanda N
  • Department of Animal and Range Sciences, Montana State University, Bozeman, MT 59717.

MeSH Terms

  • Animals
  • Horses
  • Male
  • Female
  • Lipopolysaccharides / pharmacology
  • Lipopolysaccharides / administration & dosage
  • Inflammation / veterinary
  • Inflammation / chemically induced
  • Inflammation / drug therapy
  • Horse Diseases / drug therapy
  • Horse Diseases / chemically induced
  • Clodronic Acid / pharmacology
  • Clodronic Acid / administration & dosage
  • Clodronic Acid / therapeutic use
  • Biomarkers / metabolism
  • Injections, Intra-Articular / veterinary
  • Bone Density Conservation Agents / pharmacology
  • Cartilage / metabolism
  • Cartilage / drug effects
  • Cartilage, Articular / drug effects
  • Cartilage, Articular / metabolism

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