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THU0056 Free fatty acids promote inflammation via osteoblasts and osteoclasts from patients with rheumatoid arthritis or osteoarthritis
  1. K Frommer1,
  2. A Schäffler2,
  3. U Lange1,
  4. S Rehart3,
  5. J Steinmeyer4,
  6. M Rickert4,
  7. U Müller-Ladner1,
  8. E Neumann1
  1. 1Dept. of Internal Medicine and Rheumatology, Kerckhoff-Klinik, Justus-Liebig-University of Giessen, Bad Nauheim
  2. 2Dept. of Internal Medicine III, Endocrinology, Diabetes, Metabolism, Justus-Liebig-University of Giessen, Giessen
  3. 3Dept. of Orthopaedics and Trauma Surgery, Agaplesion Markus Hospital, Frankfurt
  4. 4Dept. of Orthopaedics and Orthopaedic Surgery, University Hospital Giessen and Marburg, Giessen, Germany

Abstract

Background Various inflammatory cardiovascular and metabolic diseases such as atherosclerosis, coronary heart diseases and type 2 diabetes are associated with chronically elevated free fatty acid (FFA) levels. With inflammation being a factor in pathological bone loss, FFA may also be contributors to bone loss in osteoarthritis (OA) and/or rheumatoid arthritis (RA).

Objectives To investigate whether FFA have an influence on osteoblasts and osteoclasts from patients with RA or OA, in a way that may alter bone degradation in these diseases.

Methods Primary osteoblasts (OB) were isolated from cancellous bone of OA and RA patients undergoing knee joint surgery. Osteoclasts (OC) were differentiated from peripheral blood mononuclear cells (PBMC). OB and OC were stimulated with the saturated FFA palmitic acid (PA) and the unsaturated FFA linoleic acid (LA) (100 μM each). Immunoassays were used to quantify protein secretion. mRNA expression levels were quantified by real-time PCR. Mineralization activity was quantified using Alizarin Red S staining, differentiated OC were quantified by counting TRAP-positive multinuclear cells (>2 nuclei). Toll-like receptor (TLR) 4 and TLR2 were blocked by neutralizing antibodies.

Results When stimulated with FFA, OB from RA and OA patients secreted higher amounts of the proinflammatory cytokine IL-6 (up to 9-fold) and the chemokines IL-8 (up to 221-fold), GRO-a (from below detection level to detectable levels) and MCP-1 (up to 16-fold). Differences in the degree of response were more dependent on the patient than the disease. RANKL as well as OPG, OB-secreted modulators of OC differentiation, as well as OB differentiation markers (e.g. osterix, osteocalcin) were not influenced by FFA on mRNA or protein level. The effect of FFA on mineralization activity of OB varied between patients, yet overall there was no significant difference between FFA-treated and untreated OB. Expression of the two Wnt signaling molecules, axin-2 and b-catenin, was not changed by PA or LA, suggesting no involvement of the Wnt signaling pathway in the effects observed by FFA in OB. On the other hand, TLR4 blockade significantly reduced PA-induced IL-8 secretion by OB (by 93%), while blocking TLR2 had no effect. In both RA and OA OC, IL-8 secretion was significantly enhanced by PA and LA, with a clear time-dependency within the differentiation process for RA OC but not for OA OC. The number of TRAP positive multinuclear cells decreased for RA OC by approx. 50%, which was in agreement with the reduced TRAP secretion by a factor of 2–3 at d14. mRNA expression of various osteoclast activity markers (CLCN7, CTSK, TCIRG) was not altered.

Conclusions Inflammation is promoted by FFA via both OB and OC from patients with RA or OA, thus possibly indirectly contributing to bone loss while no direct effect on OB/OC activity could be observed. In OB, these effects are probably mainly mediated by TLR4, while TLR2 and Wnt pathways do not play a role.

Disclosure of Interest None declared

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