Objective. To delineate the expression of transforming growth factor ␤-inducible gene h3 (␤IG-H3) in rheumatoid synovitis and to determine the regulatory role of ␤IG-H3 in the adhesion and migration of fibroblast-like synoviocytes (FLS).Methods. Synovial tissue was obtained from patients with rheumatoid arthritis (RA) during joint replacement surgery, and FLS were isolated using enzymatic treatment. Immunohistochemical staining was performed to show the expression of ␤IG-H3 within rheumatoid synovium. Synthesis of ␤IG-H3 from FLS was determined by semiquantitative reverse transcription-polymerase chain reaction, Western blot analysis, and enzyme-linked immunosorbent assay. Cell adhesion and migration assays were performed using the YH18 peptide in the fourth fas-1 domain of ␤IG-H3 and function-blocking antibodies to integrins.Results. Expression of ␤IG-H3 was up-regulated in RA synovial tissue compared with synovial tissue from patients with osteoarthritis. FLS isolated from RA synovial tissue constitutively produced ␤IG-H3, which was up-regulated by transforming growth factor ␤1, interleukin-1␤, and tumor necrosis factor ␣. Although FLS expressed a variety of integrins, ␤IG-H3 mediated adhesion and migration of FLS through interaction with ␣v␤3 integrin. Cytokines failed to affect the ␤IG-H3-mediated adhesion. However, migration of FLS guided by ␤IG-H3 was enhanced by interferon-␥ and plateletderived growth factor type BB. The YH18 peptide in the fourth fas-1 domain of ␤IG-H3 inhibited adhesion and migration in a dose-dependent manner. Conclusion.The results suggest that ␤IG-H3, which is abundantly expressed in RA synovial tissue, plays a regulatory role in chronic destructive inflammation through the modulation of the adhesion and migration of FLS. This finding indicates the relevance of ␤IG-H3 and ␣v␤3 integrin-interacting motifs as potential therapeutic targets in this disease.

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Objective. Transforming growth factor ␤-inducible gene h3 (␤IG-H3), which is abundantly expressed in rheumatoid synovium, and matrix metalloproteinases (MMP) play important roles in the pathogenesis of rheumatoid arthritis (RA). The aim of this study was to determine the therapeutic efficacy of ␤IG-H3-derived peptides using MMP-1-dependent target tissue delivery in chronic inflammatory arthritis.Methods. Peptides developed from ␤IG-H3 derivatives, including the second and fourth YH peptides, the fourth fas-1 domain, the fourth fas-1 domain truncated for H1 and H2 sequences (dhfas-1), and an MMP-1-cleavable composite peptide (MFK24), were cloned. We confirmed the specificity of MFK24 cleavage by immunoblot analysis after treatment with different proteases.Results. The YH18 peptide in the fourth fas-1 domain of ␤IG-H3 was weakly effective in suppressing arthritis severity in mice with collagen-induced arthritis (CIA). Treatment with higher-dose dhfas-1 (30 mg/kg) showed remarkable efficacy, whereas treatment with a lower dose (10 mg/kg) resulted in only partial improvement. MFK24, a composite peptide consisting of dhfas-1 and RGD peptide linked by MMP-1 substrate, was cleaved specifically by MMP-1. The adhesion and migration of NIH3T3 cells mediated by ␤IG-H3 were inhibited by MFK24 at a low concentration. MFK24 suppressed the adhesion of NIH3T3 cells more efficiently compared with murine dhfas-1 (MFK00) or RGD, either alone or in combination. The therapeutic efficacy of MFK24 in mice with CIA was remarkably enhanced, with consistently reduced expression of inflammatory mediators within joint tissue.Conclusion. This proof-of-concept study showed that an MMP-cleavable composite peptide, based on ␤IG-H3 derivatives, had markedly improved therapeutic efficacy in chronic inflammatory arthritis, implicating a new expandable strategy for enhancement of the efficacy of 2 different active molecules in RA.Rheumatoid arthritis (RA) is a chronic destructive arthritis characterized by proliferating pannus, in which synoviocyte proliferation, increased expression of extracellular matrix (ECM) proteins, immune cell infiltration, and neovascularization are characteristic findings (1-3). Fibroblast-like synoviocytes (FLS) serve as one of the primary players in joint destruction by producing proteolytic enzymes, including matrix metalloproteinases (MMPs) (4), via interactions with abundantly expressed ECM proteins (5-7) and inflammatory cytokines (8).Transforming growth factor ␤-inducible gene h3 (␤IG-H3) is an ECM protein that is secreted by several cell types, such as FLS, in synovial tissue and is highly expressed in rheumatoid synovium (9,10). The ␤IG-H3 protein, which is composed of 4 homologous fas-1 domains and an Arg-Gly-Asp (RGD) motif at the carboxyl terminal,

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We demonstrated that arthritis could be visualized noninvasively using hydrophobically modified glycol chitosan nanoparticles labeled with Cy5.5 (HGC-Cy5.5) and an optical imaging system. Activated macrophages expressing Mac-1 molecules effectively phagocytosed HGC-Cy5.5, which formed spherical nanoparticles under physiologic conditions. We estimated the applicability of HGC-Cy5.5 to quantitative analysis of arthritis development and progression. Near-infrared fluorescence images, captured after HGC-Cy5.5 injection in mice with collagen-induced arthritis, showed stronger fluorescence intensity in the active arthritis group than in the nonarthritis group. According to the progression of arthritis in both collagen-induced arthritis and collagen antibody-induced arthritis models, total photon counts (TPCs) increased in parallel with the clinical arthritis index. Quantitative analysis of fluorescence after treatment with methotrexate showed a significant decrease in TPC in a dose-dependent manner. Histologic evaluation confirmed that the mechanism underlying selective accumulation of HGC-Cy5.5 within synovitis tissues included enhanced phagocytosis of the probe by Mac-1-expressing macrophages as well as enhanced permeability through leaky vessels. These results suggest that optical imaging of arthritis using HGC-Cy5.5 can provide an objective measurement of disease activity and, at the same time, therapeutic responses in rheumatoid arthritis.

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Identification of the structure-function relationship of heparin, particularly between 2-O-, 6-O-, and N-sulfation and its anticoagulant or anti-inflammatory activities, is critical in order to evaluate the biological effects of heparin, especially in conjunction with modifications for oral formulation. In this study, we demonstrated that removal of 2-O, 6-O, or N-desulfation and their hydrophobic modifications have differential effects on the blocking of interactions between sLeX and P-and L-selectins, with highest inhibition by 6-O desulfation, which was consistent with their in vivo therapeutic efficacies on CIA mice. The 6-O desulfation of lower molecular weight heparin (LMWH) retained the ability of LMWH to interfere with T cell adhesion via selectin-sLeX interactions. Furthermore, 6DSHbD coated on the apical surface of inflamed endothelium directly blocked the adhesive interactions of circulating T cells, which was confirmed in vivo by suppressing T cell adhesion at post-capillary venular endothelium. Thus, in series with our previous study demonstrating inhibition of transendothelial migration, oral delivery of low anticoagulant LMWH to venular endothelium of inflamed joint tissues ameliorated arthritis by the stepwise inhibition of T cell recruitment and provides a rationale for the development of modified oral heparins as innovative agents for the treatment of chronic inflammatory arthritis.