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Effects of transforming growth factor-β and interleukin-1β on inflammatory markers of osteoarthritis in cultured canine chondrocytes

Nadja Adler DVM1, Axel Schoeniger Dipl Biochem, Dr Rer Nat2, and Herbert Fuhrmann DVM, Prof Dr Med Vet3
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  • 1 Faculty of Veterinary Medicine, Institute of Biochemistry, University of Leipzig, 04103 Leipzig, Germany.
  • | 2 Faculty of Veterinary Medicine, Institute of Biochemistry, University of Leipzig, 04103 Leipzig, Germany.
  • | 3 Faculty of Veterinary Medicine, Institute of Biochemistry, University of Leipzig, 04103 Leipzig, Germany.

Abstract

OBJECTIVE To determine effects of transforming growth factor (TGF)-β and interleukin (IL)-1β on inflammatory markers in cultured canine chondrocytes to clarify the role of these cytokines in osteoarthritis of dogs.

SAMPLE Pooled chondrocytes isolated from the stifle joints of 4 adult dogs.

PROCEDURES Chondrocytes were isolated, cultured, and frozen at −80°C. Pooled cells were incubated in medium with or without TGF-β (1 or 10 ng/mL) and subsequently stimulated with IL-1β (10 ng/mL). Concentrations of nitric oxide (NO) and prostaglandin (PG) E were measured in culture supernatants. Gene expression of matrix metalloproteinase (MMP)-3, tissue inhibitor of metalloproteinase (TIMP)-2, inducible NO synthase (iNOS), and cyclooxygenase (COX)-2 was quantified by use of real-time quantitative PCR assays.

RESULTS Stimulation with IL-1β increased gene expression of all inflammatory markers, except for TIMP-2. Incubation with TGF-β resulted in a significant decrease in MMP-3 and TIMP-2 mRNA concentrations but had no effect on PGE and NO concentrations. For cells treated with TGF-β followed by IL-1β, concentrations of PGE and NO were lower, compared with concentrations for IL-1β control cells. Furthermore, IL-1β–induced gene expression of iNOS, MMP-3, and COX-2 was downregulated. However, the IL-1β–induced downregulation of TIMP-2 gene expression was partially restored by pretreatment with TGF-β.

CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that IL-1β increased the expression of inflammatory genes and mediators, and TGF-β largely attenuated the IL-1β–mediated inflammatory response. Therefore, TGF-β might be a novel target for use in the prevention and treatment of cartilage breakdown in dogs with osteoarthritis.

Contributor Notes

Ms. Adler's present address is Kleintierpraxis und Tierärztliche Klinik für Kleintiere, Gartenstraße 12, 26122 Oldenburg, Germany.

Address correspondence to Dr. Fuhrmann (fuhrmann@vmf.uni-leipzig.de).