Session Type: ACR Poster Session C
Session Time: 9:00AM-11:00AM
The pathophysiology of osteoarthritis (OA) involves wear and tear, and a state of low-grade inflammation. Wear and tear leads to tissue degradation followed by tissue repair responses including TGFβ-induced myofibroblast production of extracellular matrix (ECM). Fibronectins are an essential part of the ECM, and injection of fibronectin fragments into rabbit joints is an established animal model of OA. Recently, alternatively spliced fibronectin containing the ED-A domain (ED-A FN) has been shown to activate Toll-like receptor 4. In this study, we hypothesize that FN fragments containing the ED-A domain could be one mechanism transducing mechanical events into inflammatory signals in OA.
Samples of synovial membrane and cartilage were obtained from patients with knee OA undergoing joint replacement surgery. Immunostaining was performed on synovial membranes. Fibroblast-like synovial cells (FLS) isolated by enzymatic digestion of remnant synovial membrane were stimulated with TGFβ, TNFα, lipopolysaccharide, IL-6, OA synovial fluid from two different donors, or chondrocyte lysate, and culture supernatants were analyzed for ED-A FN by immunofluorescence staining. ED-A FN fragments were obtained by plasmin digestion of cellular FN. Synovial cells isolated by enzymatic digestion and human monocyte-derived macrophages (MDM) were incubated with recombinant ED-A FN, plasmin, cellular FN, or cellular FN digested with plasmin; and culture supernatants were analyzed for MCP-1 and TNFα.
We hypothesized that ED-A FN is produced by OA FLS in response to products reflecting tear and wear in OA. Indeed, the production of ED-A FN by OA FLS was increased by TGFβ, OA synovial fluid, and lysed chondrocytes in all experiments (n=3, see figure). ED-A FN co-localized with the myofibroblast marker αSMA in both the OA FLS (n=3) and in the OA synovial membranes (n=8). We further hypothesized that ED-A FN expression is associated with inflammation in OA. ED-A FN staining was associated with both number of lining layer cells (rho=0.85 and p=0.011) and infiltrating sublining cells (rho=0.88 and p=0.007) in the OA synovium (n=8), and co-localized with both MCP-1 and TNFα (n=5). Recombinant ED-A FN increased the production of both MCP-1 and TNFα by MDM (n=3) and OA FLS (n=3). Finally, we demonstrated that the FN fragments containing the ED-A domain generated the same production of both MCP-1 and TNFα as recombinant ED-A FN.
The disease process in OA shares features with the chronic wound healing response including myofibroblast differentiation and that humeral mediators found in the joint can promote myofibroblast production of ED-A FN. We additionally show that recombinant and plasmin-derived ED-A fragments can induce generation of pro-inflammatory mediators from FLS and MDM. This study supports targeting the formation of ED-A FN or the enzymatic fragmentation of FN to reduce pro-inflammatory responses in OA.
To cite this abstract in AMA style:Kragstrup TW, Sohn DH, Lepus C, Onuma K, Wang Q, Robinson WH, Sokolove J. Fibroblast-like Synovial Cell Production of ED-a Fibronectin Contributes to Inflammation in Osteoarthritis [abstract]. Arthritis Rheumatol. 2017; 69 (suppl 10). https://acrabstracts.org/abstract/fibroblast-like-synovial-cell-production-of-ed-a-fibronectin-contributes-to-inflammation-in-osteoarthritis/. Accessed September 24, 2021.
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ACR Meeting Abstracts - https://acrabstracts.org/abstract/fibroblast-like-synovial-cell-production-of-ed-a-fibronectin-contributes-to-inflammation-in-osteoarthritis/