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Equine veterinary journal2022; 55(2); 325-335; doi: 10.1111/evj.13582

Conditioned serum in vitro treatment of chondrocyte pellets and osteoarthritic explants.

Abstract: Autologous conditioned serum (ACS) is used to treat osteoarthritis in horses, although its effects are not fully investigated. Objective: To investigate the effects of equine serum and conditioned serum on chondrocytes stimulated with interleukin (IL)-1β and cartilage explants with mild osteoarthritis. Methods: In vitro experimental study. Methods: The effect of three different serum preparations (unincubated control [PS], serum incubated 24 h [PS24h] and serum incubated 24 h in ACS containers [PCS]) pooled from lame horses were tested in two in vitro models. IL-1β and IL-1 receptor antagonist (IL-1Ra) concentrations were measured in all sera. In model 1, chondrocyte pellet cultures were stimulated with IL-1β prior to treatment with the serum preparations for 2 and 48 h. Microarray, polymerase chain reaction, and matrix metallopeptidase-13 analyses were performed. In model 2, cartilage explants from horses with structural osteoarthritis were treated with PS or PCS on days 0, 6 and 12, or left untreated, and evaluated at day 24 using the OARSI grading scale for histological evaluation of articular cartilage. Results: The IL-1Ra concentration in PS24h and PCS was significantly higher than in PS. In model 1, inflammation- and cartilage matrix degradation-related genes were upregulated after 48 h in all treatment groups versus untreated controls. Cartilage matrix molecules, aggrecan and collagens, were downregulated in PS24h- and PCS-treated pellets versus untreated controls. Growth factor signalling genes were upregulated-FGF7 in all treatment groups, BMP2 in PS24h-, and INHBA in PCS-treated-compared with untreated controls. In model 2, the OARSI score at day 24 was not significantly different between treatment groups. Conclusions: Results from in vitro models cannot be directly translated to in vivo situations. Conclusions: In vitro treatment with conditioned serum did not alleviate IL-1β-induced responses in chondrocyte pellets or lead to morphological improvement in osteoarthritic cartilage explants. Unassigned: Suero autólogo acondicionado (ACS) es usado para tartar osteoartritis en caballos, aunque sus efectos no han sido completamente investigados. Objective: Investigar los efectos de suero equino y suero acondicionado en condrocitos estimulados con interleukina (IL)-1β y explantes de cartílago con osteoartritis leve. DISEÑO DEL ESTUDIO: Estudio experimental in vitro. MÉTODOS: El efecto de tres preparaciones séricas diferentes (control no incubado (PS), suero incubado 24 h (PS24h), y suero incubado 24 h en frascos ACS (PCS)) combinados y obtenidos de caballos cojos fueron probados en dos modelos in vitro. Las concentraciones de IL-1β y de receptor antagonista de IL-1 (IL-1Ra) fueron medidas en todos los sueros. En el modelo 1, los cultivos de pellets de condrocitos fueron estimulados con IL-1β antes de ser tratados con las preparaciones séricas durante 2 y 48 h. Se realizaron análisis de micromatrices, reacciones de polimerasa en cadena y de matriz de metalopeptidasa-13. En el modelo 2, explantaciones de cartílago proveniente de caballos con osteoartritis estructural fueron tratados con PS o PCS en los días 0, 6 y 12, o dejados sin tartar, y evaluados al día 24 usando la escala de graduación OARSI para evaluación histológica de cartílago articular. Results: La concentración de IL-1Ra en PS24h y PCS fue significativamente mayor que en PS. En el modelo 1, los genes relacionados a la inflamación y a la degradación de la matriz cartilaginosa estaban aumentados después de 48 h en todos los grupos tratados en comparación a los controles no tratados. Las moléculas de matriz cartilaginosa, agrecanos y colágenos estaban disminuidos en los pellets PS24h y PCS versus los controles no tratados. Los genes de señales de factores de crecimiento FGF7 estaban aumentados en todos los grupos tratados, BMP2 en PS24h y INHBA in PCS en comparación con los controles no tratados. En el modelo 2, la escala OARSI al día 24 no fue significativamente distinta entre los grupos de tratamientos. Unassigned: Los resultados de modelos in vitro no pueden ser directamente aplicados a situaciones in vivo. Conclusions: El tratamiento in vitro con suero acondicionado no alivió las respuestas inducidas por IL-1β en pellets de condrocitos o llevo a mejoramiento morfológico en explantes de cartílago con osteoartritis.
© 2022 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.
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Publication Date: 2022-06-06 PubMed ID: 35514185PubMed Central: PMC10084271DOI: 10.1111/evj.13582Google 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 investigates the treatment effects of autologous conditioned serum (ACS), specifically incubated horse serum, on osteoarthritic chondrocytes and cartilage explants. The in vitro study revealed no notable improvements in the inflammation response or the structure of osteoarthritic tissue after ACS treatment.

Research Objective and Methods

  • The study’s aim was to understand the impact of equine serum and conditioned serum on chondrocytes (cells in the cartilage) that have been stimulated with interleukin (IL)-1β, a pro-inflammatory cytokine, and cartilage explants (fragments) displaying mild osteoarthritis.
  • This was an in vitro experimental study that made use of three different types of serum preparations. The serums were pooled from lame horses. The serum types included unincubated control (PS), serum incubated for 24 hours (PS24h), and serum incubated for the same period in ACS containers (PCS).
  • The concentrations of IL-1β and IL-1 receptor antagonist (IL-1Ra) were measured in all serums. This was done to check for necessary proteins for counteracting inflammatory responses.

    • Experimental Models and Analysis

    • The first experimental model involved treating chondrocyte pellet cultures with IL-1β to stimulate an inflammatory response before subjecting them to the serum treatments. After specified periods, reactions including microarray (gene activity tests), polymerase chain reaction (DNA tests), and matrix metalloproteinase-13 (a protein-degrading enzyme) were monitored.
    • In the second model, cartilage explants taken from horses showing signs of osteoarthritis were treated with PS or PCS serum at specific intervals or left untreated. The severity of osteoarthritis and the effects of the treatment were evaluated using the OARSI grading scale for histological evaluation of articular cartilage.

      • Results and Conclusion

      • In both trial models, IL-1Ra concentration was significantly higher in PS24h and PCS compared to PS.
      • In the chondrocyte pellet cultures (Model 1), genes related to inflammation and cartilage matrix degradation were elevated after 48 hours in all treatment groups, compared to the untreated controls. Cartilage matrix molecules, like aggrecan and collagen, were less abundant in the treatments with PS24h and PCS compared to untreated controls.
      • In the cartilage explant trial (Model 2), the OARSI score at day 24 showed no significant differences between the treatment groups, suggesting no notable improvement in osteoarthritis severity.
      • The study concluded that the in vitro treatment with conditioned serum did not alleviate IL-1β-induced responses in the chondrocyte pellets or lead to any morphological improvement in the osteoarthritic cartilage explants.

Cite This Article

APA
Löfgren M, Ekman S, Ekholm J, Engström M, Fjordbakk CT, Svala E, Holm Forsström K, Lindahl A, Skiöldebrand E. (2022). Conditioned serum in vitro treatment of chondrocyte pellets and osteoarthritic explants. Equine Vet J, 55(2), 325-335. https://doi.org/10.1111/evj.13582

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 55
Issue: 2
Pages: 325-335

Researcher Affiliations

Löfgren, Maria
  • Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Ekman, Stina
  • Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Ekholm, Josefine
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.
Engström, Mona
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.
Fjordbakk, Cathrine T
  • Faculty of Veterinary Medicine, Department of Companion Animal Clinical Sciences, Norwegian University of Life Sciences, Oslo, Norway.
Svala, Emilia
  • Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden.
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.
Holm Forsström, Karin
  • Equine Clinic, University Animal Hospital, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Lindahl, Anders
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.
Skiöldebrand, Eva
  • Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden.
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.

MeSH Terms

  • Horses
  • Animals
  • Chondrocytes / metabolism
  • Interleukin 1 Receptor Antagonist Protein / metabolism
  • Interleukin 1 Receptor Antagonist Protein / pharmacology
  • Osteoarthritis / therapy
  • Osteoarthritis / veterinary
  • Cartilage, Articular
  • Inflammation / metabolism
  • Inflammation / veterinary
  • Cells, Cultured
  • Horse Diseases / metabolism

Grant Funding

  • Valborg Jacobssons Fond
  • Svelands Stiftelse
  • H-16-47-182 / Swedish-Norwegian Foundation for Equine Research

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 4 times.
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