ARTHRITIS & RHEUMATISM Vol. 62, No. 3, March 2010, pp 815–825 DOI 10.1002/art.27295 © 2010, American College of Rheumatology Trefoil Factor 3 Is Induced During Degenerative and Inflammatory Joint Disease, Activates Matrix Metalloproteinases, and Enhances Apoptosis of Articular Cartilage Chondrocytes Sophie Ro ¨sler, 1 Tobias Haase, 1 Horst Claassen, 1 Ute Schulze, 1 Martin Schicht, 1 Dagmar Riemann, 1 Jo ¨rg Brandt, 1 David Wohlrab, 1 Brigitte Mu ¨ller-Hilke, 2 Mary B. Goldring, 3 Saadettin Sel, 1 Deike Varoga, 4 Fabian Garreis, 1 and Friedrich P. Paulsen 1 Objective. Trefoil factor 3 (TFF3, also known as intestinal trefoil factor) is a member of a family of protease-resistant peptides containing a highly con- served motif with 6 cysteine residues. Recent studies have shown that TFF3 is expressed in injured cornea, where it plays a role in corneal wound healing, but not in healthy cornea. Since cartilage and cornea have similar matrix properties, we undertook the present study to investigate whether TFF3 could induce anabolic functions in diseased articular cartilage. Methods. We used reverse transcriptase– polymerase chain reaction, Western blot analysis, and immunohistochemistry to measure the expression of TFF3 in healthy articular cartilage, osteoarthritis (OA)–affected articular cartilage, and septic arthritis– affected articular cartilage and to assess the effects of cytokines, bacterial products, and bacterial superna- tants on TFF3 production. The effects of TFF3 on matrix metalloproteinase (MMP) production were mea- sured by enzyme-linked immunosorbent assay, and ef- fects on chondrocyte apoptosis were studied by caspase assay and annexin V assay. Results. Trefoil factors were not expressed in healthy human articular cartilage, but expression of TFF3 was highly up-regulated in the cartilage of pa- tients with OA. These findings were confirmed in animal models of OA and septic arthritis, as well as in tumor necrosis factor – and interleukin-1–treated primary human articular chondrocytes, revealing induction of Tff3/TFF3 under inflammatory conditions. Application of the recombinant TFF3 protein to cultured chondro- cytes resulted in increased production of cartilage- degrading MMPs and increased chondrocyte apoptosis. Conclusion. In this study using articular cartilage as a model, we demonstrated that TFF3 supports cata- bolic functions in diseased articular cartilage. These findings widen our knowledge of the functional spec- trum of TFF peptides and demonstrate that TFF3 is a multifunctional trefoil factor with the ability to link inflammation with tissue remodeling processes in artic- ular cartilage. Moreover, our data suggest that TFF3 is a factor in the pathogenesis of OA and septic arthritis. The mammalian trefoil factor (TFF) family com- prises 3 protease-resistant peptides of 7–12 kd (TFF1 [formerly called pS2], TFF2 [formerly called hSP], and TFF3 [formerly called hP1.B/hITF or ITF]) that have in common a distinct highly conserved motif of 6 cysteine Supported in part by the GI Company. Dr. Goldring’s work was supported by the NIH (grant R01-AG-022021). Dr. Paulsen’s work was supported by the DFG (grants PA 738/9-1, PA 738/9-2, and PA 738/6-1) and the Bundesministerium fu ¨r Bildung und Forschung– Wilhelm Roux Program, Halle, Germany (grants FKZ 09/17, FKZ 14/24, and FKZ 14/25). 1 Sophie Ro ¨sler, Tobias Haase, DiplBiochem, Horst Claassen, MD, Ute Schulze, MSc, Martin Schicht, MSc, Dagmar Riemann, MD, Jo ¨rg Brandt, MD, David Wohlrab, MD, Saadettin Sel, MD, Fabian Garreis, MSc, Friedrich P. Paulsen, MD: Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany; 2 Brigitte Mu ¨ller-Hilke, PhD: University of Rostock, Rostock, Germany; 3 Mary B. Goldring, PhD: Hospital for Special Surgery, Weill College of Medicine of Cornell University, New York, New York; 4 Deike Varoga, MD: Christian Albrecht University, Kiel, Germany. Ms Ro ¨sler and Mr. Haase contributed equally to this work. Dr. Wohlrab has received speaking fees from Smith & Nephew (less than $10,000). Address correspondence and reprint requests to Friedrich P. Paulsen, MD, Department of Anatomy and Cell Biology, Martin Luther University Halle-Wittenberg, Faculty of Medicine, Grosse Steinstrasse 52, D-06097 Halle (Saale), Germany. E-mail: friedrich.paulsen@medizin.uni-halle.de. Submitted for publication July 4, 2009; accepted in revised form November 17, 2009. 815