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BMC veterinary research2024; 20(1); 546; doi: 10.1186/s12917-024-04403-2

Differential modulation of inflammatory cytokines by recombinant IL-10 in IL-1β and TNF-α ̶ stimulated equine chondrocytes and synoviocytes: impact of washing and timing on cytokine responses.

Abstract: Osteoarthritis (OA) remains a challenging joint disorder necessitating effective anti-inflammatory interventions. In this study, our primary objective was to establish an in vitro protocol that replicates the clinical investigation of anti-inflammatory drugs intended for OA management. Focusing on recombinant IL-10 (r.IL-10) as a potential anti-inflammatory treatment, we designed and implemented two distinct protocols to evaluate the efficacy of r.IL-10 in modulating chondrocyte and synoviocyte inflammation.The experimental design involved sequential stimulation with IL-1β and TNF-α for 24 h, followed by washing (model 1) or not washing (model 2) the cells before r.IL-10 treatment. Samples were collected after 6-24 h of treatment. Cellular responses were evaluated by quantifying gene expression and synthesis of key inflammatory cytokines and proteases.The expression and synthesis of inflammatory cytokines and proteases was significantly affected by washing and treatment time. The expression of IL-1β, TNF-α, IL-8, MMP-13, and ADAMTS5 were effectively reduced in r.IL-10-treated chondrocytes and synoviocytes in model 2 after 24 h, particularly at concentrations of 10 and 20 ng/mL. r.IL-10 treatment significantly increased IL-6 gene expression in chondrocytes at all time points. However, in synoviocytes, IL-6 expression was significantly lower in model 2 after 24 h of r.IL-10 treatment. r.IL-10 treatment significantly decreased IL-1β and TNF-α content in synoviocyte supernatants, particularly in model 2 at concentrations of 10 and 20 ng/mL after 6 and 24 h. r.IL-10 treatment in chondrocytes led to a significant decrease in IL-1β supernatant concentrations in model 2 after 24 h only.This study demonstrated that r.IL-10 treatment effectively reduces key inflammatory markers and matrix metalloproteinase activity in both chondrocytes and synoviocytes, particularly in model 2 where cells were not washed prior to treatment. These findings highlight r.IL-10's potential as a robust anti-inflammatory agent for OA management and suggest its critical role in developing effective therapeutic strategies for OA.
© 2024. The Author(s).
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Publication Date: 2024-12-02 PubMed ID: 39623412PubMed Central: PMC11610082DOI: 10.1186/s12917-024-04403-2Google Scholar: Lookup
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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 investigated the modulatory effects of a potential anti-inflammatory treatment, r.IL-10, on inflammation caused by osteoarthritis (OA) in connective tissues. Two protocols were used; one involving washing cells prior to treatment, and the other without washing. The study found that r.IL-10 was effective in reducing inflammation and proteases involved in tissue breakdown, particularly when cells were not washed prior to treatment.

Research Aim and Methodology

  • The study aimed to establish an in vitro protocol that could simulate the clinical process of investigating anti-inflammatory drugs for OA treatment. The focus was on the differential effects of the recombinant IL-10 (r.IL-10) as an anti-inflammatory treatment.
  • The investigation involved two models: In model 1, cells were sequentially stimulated with IL-1β and TNF-α, two molecules involved in inflammatory response, for 24 hours. After this, the cells were washed before application of r.IL-10 treatment. In model 2, the same process was followed but the cells were not washed before r.IL-10 treatment.
  • Following r.IL-10 treatment, samples were collected every 6-24 hours. The cell responses to the treatment were evaluated by measuring the expression of key inflammatory genes and the production of inflammatory cytokines and proteases.

Key Findings

  • The results showed that washing and treatment time significantly affected the expression and production of inflammatory cytokines and proteases.
  • The expression of IL-1β, TNF-α, IL-8, MMP-13, and ADAMTS5 – molecules that trigger inflammation and tissue breakdown – were effectively reduced with the r.IL-10 treatment, particularly after 24 hours in model 2.
  • r.IL-10 increased IL-6 gene expression in chondrocytes at all time points which may denote an anti-inflammatory response, but decreased IL-6 expression in synoviocytes specifically in model 2 after 24h of treatment.
  • r.IL-10 treatment led to a significant decrease in IL-1β and TNF-α content in synoviocytes, particularly in model 2 at concentrations of 10 and 20 ng/mL after 6 and 24h, indicating its anti-inflammatory effect.
  • r.IL-10 treatment in chondrocytes led to a significant decrease in IL-1β supernatant concentrations in model 2 after 24h.

Conclusion

  • The study concluded that r.IL-10 treatment can effectively reduce key inflammatory markers and matrix metalloproteinase activity. Precisely, when cells were not washed before treatment (model 2), r.IL-10 treatment had superior anti-inflammatory effects, demonstrating possible therapeutic potential for OA management.

Cite This Article

APA
Elkhenany HA, Linardi RL, Ortved KF. (2024). Differential modulation of inflammatory cytokines by recombinant IL-10 in IL-1β and TNF-α ̶ stimulated equine chondrocytes and synoviocytes: impact of washing and timing on cytokine responses. BMC Vet Res, 20(1), 546. https://doi.org/10.1186/s12917-024-04403-2

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 20
Issue: 1
Pages: 546
PII: 546

Researcher Affiliations

Elkhenany, Hoda A
  • Department of Clinical Studies-New Bolton Center, University of Pennsylvania, 382 West Street Road, Kennett Square, PA, 19348, USA.
  • Department of Surgery, Faculty of Veterinary Medicine, Alexandria University, Alexandria, Egypt.
Linardi, Renata L
  • Department of Clinical Studies-New Bolton Center, University of Pennsylvania, 382 West Street Road, Kennett Square, PA, 19348, USA.
Ortved, Kyla F
  • Department of Clinical Studies-New Bolton Center, University of Pennsylvania, 382 West Street Road, Kennett Square, PA, 19348, USA. kortved@vet.upenn.edu.

MeSH Terms

  • Animals
  • Chondrocytes / drug effects
  • Chondrocytes / metabolism
  • Horses
  • Interleukin-1beta / metabolism
  • Interleukin-1beta / pharmacology
  • Synoviocytes / drug effects
  • Synoviocytes / metabolism
  • Tumor Necrosis Factor-alpha / metabolism
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / pharmacology
  • Interleukin-10 / metabolism
  • Interleukin-10 / genetics
  • Cytokines / metabolism
  • Cytokines / genetics
  • Recombinant Proteins / pharmacology
  • Cells, Cultured
  • Anti-Inflammatory Agents / pharmacology

Conflict of Interest Statement

Declarations. Ethics approval and consent to participate: The study involving equine cartilage and synovium tissue collection was approved by the Institutional Animal Care and Use Committee (IACUC), the University of Pennsylvania (Animal Use Protocol No. #806625). Consent for publication: All authors have read and agreed to the published version of the manuscript. Competing interests: The authors declare no competing interests.

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Citations

This article has been cited 2 times.
  1. Shefa MS, Kim W. The Role of SLPI Gene-Mediated Inflammation in Osteoarthritis. Biomolecules 2025 Jun 20;15(7).
    doi: 10.3390/biom15070909pubmed: 40723781google scholar: lookup
  2. Kim SH, Yoon SM, Ahn JH, Choi YJ. Effect of Remimazolam and Propofol on Blood Glucose and Serum Inflammatory Markers in Patients with Type 2 Diabetes: A Clinical Trial with Prospective Randomized Control. Medicina (Kaunas) 2025 Mar 17;61(3).
    doi: 10.3390/medicina61030523pubmed: 40142333google scholar: lookup
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