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Research article - Peer-reviewed, 2022

Conditioned serum in vitro treatment of chondrocyte pellets and osteoarthritic explants

Lofgren, Maria; Ekman, Stina; Ekholm, Josefine; Engstrom, Mona; Fjordbakk, Cathrine T.; Svala, Emilia; Forsstrom, Karin Holm; Lindahl, Anders; Skioldebrand, Eva

Abstract

Background: Autologous conditioned serum (ACS) is used to treat osteoarthritis in horses, although its effects are not fully investigated.Objectives: To investigate the effects of equine serum and conditioned serum on chondrocytes stimulated with interleukin (IL)-1 beta and cartilage explants with mild osteoarthritis.Study design: 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-1 beta and IL-1 receptor antagonist (IL-1Ra) concentrations were measured in all sera. In model 1, chondrocyte pellet cultures were stimulated with IL-1 beta 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.Main limitations: Results from in vitro models cannot be directly translated to in vivo situations.Conclusions: In vitro treatment with conditioned serum did not alleviate IL-1 beta-induced responses in chondrocyte pellets or lead to morphological improvement in osteoarthritic cartilage explants.

Keywords

autologous conditioned serum; horse; IL-1 receptor antagonist; inflammation; interleukin 1 beta; osteoarthritis

Published in

Equine Veterinary Journal
2022,
Publisher: WILEY