Inhibition of monosodium urate-induced peritoneal neutrophil influx by anti-IL-1 treatment

<p><b>Copyright information:</b></p><p>Taken from "A pilot study of IL-1 inhibition by anakinra in acute gout"</p><p>http://arthritis-research.com/content/9/2/R28</p><p>Arthritis Research & Therapy 2007;9(2):R28-R28.</p><p>Published online 12 Mar 2007</p><p>PMCID:PMC1906806.</p><p></p> BALB/C mice were injected intraperitoneally with 0.5 mg monosodium urate (MSU) crystals together with PBS or anti-IL-1RI mAb (200 μg) or anakinra (200 μg). BALB/C mice were injected intraperitoneally with 0.5 mg MSU crystals together with PBS or anti-TNF mAb (200 μg) or anakinra (200 μg). Neutrophil influx in the peritoneum was quantified 6 hours later. Values are the mean ± standard error of the mean of five mice per group. An unpaired Student's test was used to calculate the value

STS inhibits chondrocyte IL-6 and MCP-1 secretion in a dose-dependent manner.

<p>(<b>a</b>) IL-6 and MCP-1 secretion by primed murine joint chondrocytes treated or not with 25mM STS for 1, 3 or 7 days. Values represent means±SD of triplicates from one representative experiment of three independent experiments. (<b>b</b>) qRT-PCR of IL-6 gene in primed murine chondrocytes treated or not with 25 mM STS for 3 days. (<b>c</b>) IL-6 and MCP-1 secretion by primed murine joint chondrocytes treated or not with different concentrations of STS for 3 and 7 days. Values represent means±SD of triplicates from one experiment. *<i>p</i><0.05, ** <i>p</i><0.01, *** <i>p</i><0.001, **** <i>p</i><0.0001.</p

STS inhibits HA-induced ROS production and enzymes involved in cartilage degradation.

<p>ROS generation by (<b>a</b>) murine joint chondrocytes and (<b>b</b>) BMDM stimulated or not with HA crystals (500ug/ml) and treated or not with 25mM STS for 1h. Dihydroethidium (DHE) staining was used as an indicator of intracellular ROS generation. MitoSOX^™ Red reagent was used as an indicator of mitochondrial ROS generation. Values represent means±SD of triplicates from one representative experiment of two independent experiments and are expressed as %of ROS in unstimulated cells. (<b>c</b>) qRT-PCR of the indicated genes in murine chondrocytes stimulated or not with HA crystals (500ug/ml) and treated or not with 25mM STS for 30 minutes in DMEM. Values represent means±SD of triplicate samples. *<i>p</i><0.05, ** <i>p</i><0.01, *** <i>p</i><0.001, **** <i>p</i><0.0001.</p

STS inhibits murine joint chondrocytes mineralization.

<p>(<b>a</b>) Alizarin red and Von Kossa staining of murine chondrocytes treated or not (Nt) with 25mM STS for 7 days in complete BGJb. Pictures represent one representative culture well of one experiment (five independent experiments were performed). Scale bar 100μm. The graph shows Alizarin red absorbance at 405nm, expressed in % over Nt cells at 1, 3 or 7 days of culture. Values represent means±SD of triplicates from one representative experiment of three independent experiments. (<b>b</b>) Percentage of cytotoxicity in murine chondrocytes treated or not with 25mM STS for 1, 3 or 7 days. Values represent means±SD of triplicates from one representative experiment of three independent experiments. (<b>c</b>) Alizarin red staining of murine chondrocytes treated or not (Nt) with different concentrations of STS for 7 days. Pictures represent triplicates from one experiment. Scale bar 100μm. The graph shows Alizarin red absorbance at 405nm, expressed in % over Nt cells at 7 days of culture. Values represent means±SD of triplicates from one representative experiment of three independent experiments. (<b>d</b>) qRT-PCR of the indicated genes in murine chondrocytes treated or not with 25 mM STS for 7 days in complete DMEM. Values represent means±SD of triplicate samples. *<i>p</i><0.05, ** <i>p</i><0.01, *** <i>p</i><0.001, **** <i>p</i><0.0001. nd = not detectable</p

STS inhibits formation of new calcific deposits and cartilage degradation in experimental OA.

<p>(<b>a</b>) Representative micro-CT scan images of menisectomized murine knee joints after vehicle (PBS) or STS treatment for two months after surgery. White arrows show periarticular deposits in menisectomized knees treated with vehicle and their reduction in STS treated mice. Graphs show CTAnalyzer quantitative analysis of new formation volume (mm³) and new formation crystal content (μg) in PBS- and STS-treated menisectomized mice. Data are expressed as the mean±SD (<b>b</b>) Representative histologies of PBS- and STS-treated menisectomized knees, stained with Safranin-O. Red arrows show degenerative OA changes in the articular cartilage of PBS-treated mice. Scale bars 150 μm. Graphs show tibial and femoral scoring of cartilage damage and Safranin-O loss, accordingly to OARSI method. Values represent means±SD. (<b>c</b>) Correlation graph between tibial cartilage damage score and new formation volume of pooled PBS- and STS-treated mice. (Mice number: PBS n = 20, STS n = 19).</p

STS inhibits HA-induced cytokine and chemokine selectively in chondrocytes.

<p>(<b>a</b>) IL-6 and MCP-1 secretion by primed murine joint chondrocytes and (<b>b</b>) IL-6, MCP-1, IL-1β and TNF-α secretion by primed murine BMDM, stimulated or not with HA crystals (500ug/ml) and treated or not with 25mM STS 25mM for 2, 6 and 24hrs. Values represent means±SD of triplicates from one representative experiment of three independent experiments. (<b>c</b>) IL-6 secretion by human cartilage explants was measured by ELISA. Explants were stimulated with 500μg/ml of HA crystals in presence or absence of STS 25Mm for 24 h. Matched-halves of cartilage explants are connected by a line (5 explants for patients 1, 3 for patient 2, 4 for patient 3). Values represent means±SD of triplicate samples. *<i>p</i><0.05, ** <i>p</i><0.01, *** <i>p</i><0.001, **** <i>p</i><0.0001.</p

Essential role of platelet activation via protease activated receptor 4 in tissue factor-initiated inflammation-7

Hanges. Phosphate-buffered saline-injected mice showed minimal signs of inflammation. Staining for macrophages was strongly positive. CD3-positive T cells were also present. Staining specificity was confirmed using, as primary antibody, nonimmune isotype-matched antibodies. Fibrin deposition was assessed by fibrin immunohistochemistry.<p><b>Copyright information:</b></p><p>Taken from "Essential role of platelet activation via protease activated receptor 4 in tissue factor-initiated inflammation"</p><p>http://arthritis-research.com/content/10/2/R42</p><p>Arthritis Research & Therapy 2008;10(2):R42-R42.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2453761.</p><p></p

Essential role of platelet activation via protease activated receptor 4 in tissue factor-initiated inflammation-4

*= 0.001, Wilcoxon rank sum test. WT, wildtype; KO, knockout.<p><b>Copyright information:</b></p><p>Taken from "Essential role of platelet activation via protease activated receptor 4 in tissue factor-initiated inflammation"</p><p>http://arthritis-research.com/content/10/2/R42</p><p>Arthritis Research & Therapy 2008;10(2):R42-R42.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2453761.</p><p></p

Essential role of platelet activation via protease activated receptor 4 in tissue factor-initiated inflammation-3

Ur different protease activated receptors (PARs). PAR-1-deficient mice. PAR-2-deficient mice. PAR-3-deficient mice. PAR-4-deficient mice. In each experiment, footpad swelling was assessed in the PAR-deficient mice and their littermates (+/+ or +/-) as controls.<p><b>Copyright information:</b></p><p>Taken from "Essential role of platelet activation via protease activated receptor 4 in tissue factor-initiated inflammation"</p><p>http://arthritis-research.com/content/10/2/R42</p><p>Arthritis Research & Therapy 2008;10(2):R42-R42.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2453761.</p><p></p

Essential role of platelet activation via protease activated receptor 4 in tissue factor-initiated inflammation-2

or untreated mice (n = 7), were injected with 1 μg soluble tissue factor. Results from all treated groups were significantly reduced (< 0.05 by test) compared with the control group. Wildtype mice were treated with ancrod (n = 7) or with phosphate-buffered saline (n = 7) and then injected with 1 μg soluble tissue factor. Results from ancrod-treated mice are significantly different from the control group, at all time points (< 0.05 by test).<p><b>Copyright information:</b></p><p>Taken from "Essential role of platelet activation via protease activated receptor 4 in tissue factor-initiated inflammation"</p><p>http://arthritis-research.com/content/10/2/R42</p><p>Arthritis Research & Therapy 2008;10(2):R42-R42.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2453761.</p><p></p