Co-culture of osteochondral explants and synovial membrane as in vitro model for osteoarthritis

• DOI: 10.1371/journal.pone.0214709
• Publication Date: 2019-04-02T17:33:55Z
• Authors: Eva Haltmayer; Iris Ribitsch; Simone Gabner; Julie Rosser; Sinan Gueltekin; Johannes Peham; Ulrich Giese; Marlies Dolezal; Monika Egerbacher; Florien Jenner
• Keywords: Aggrecan; MMP3

The purpose of the current study was to establish an in vitro model for osteoarthritis (OA) by co-culture osteochondral and synovial membrane explants. Osteochondral explants were cultured alone (control-1) or with (control-2) standard culture medium interleukin-1β (IL1β) tumor necrosis factor (TNFα) added (OA-model-1 = explant; OA-model-2 osteochondroal-synovial explant). In addition, OA-model groups a 2-mm partial-thickness defect created centre cartilage explant. Changes expression extracellular matrix (ECM) genes (collagen type-1 (Col1), Col2, Col10 aggrecan) as well presence quantity inflammatory marker (IL6, metalloproteinase-1 (MMP1), MMP3, MMP13, disintegrin metalloproteinase with-thrombospondin-motif-5 (ADAMTS5) analysed immunohistochemistry, qPCR ELISA. To monitor activity classically-activated pro-inflammatory (M1) versus alternatively-activated anti-inflammatory/repair (M2) macrophages, nitric oxide/urea ratio supernatant osteochondral-synovial explant co-cultures determined. both immunohistochemistry showed significantly increased MMPs IL6 compared their respective control group. ELISA results confirmed statistically significant increase MMP1and MMP3 production over culturing period. group, indicating shift toward M1 macrophages. summary, chemical damage (TNFα, IL1β) combination elicits response similar naturally occurring OA without showing closer approximation due additional macrophages phenotype.