Quantification of aggrecan and collagen degradation products at days 7, 11, and 17

Articular cartilage explants were cultured in the presence or absence of oncostatin M plus tumour necrosis factor (OSM + TNF). Conditioned medium was collected at days 7, 11, and 17. Aggrecanase-mediated aggrecan degradation was measured by the ARGSV-G2 enzyme-linked immunosorbent assay (ELISA), matrix metalloproteinase (MMP)-mediated aggrecan degradation was quantified by the FFGVG-G2 ELISA, and MMP-mediated collagen type II degradation was quantified in the CTX-II ELISA. **< 0.01, ***< 0.001. CTX-II, crosslinked C-terminal neo-epitopes of type II collagen.<p><b>Copyright information:</b></p><p>Taken from "Cartilage degradation is fully reversible in the presence of aggrecanase but not matrix metalloproteinase activity"</p><p>http://arthritis-research.com/content/10/3/R63</p><p>Arthritis Research & Therapy 2008;10(3):R63-R63.</p><p>Published online 30 May 2008</p><p>PMCID:PMC2483454.</p><p></p

Insulin growth factor (IGF) stimulates local replenishment of cartilage

Articular cartilage explants were cultured with either oncostatin M plus tumour necrosis factor (OSM + TNF) or vehicle for 7, 11, and 17 days. Subsequently, cartilage explants were paraffin-embedded and stained for aggrecan content as described in Materials and methods. Aggrecan is completely depleted from the tissue at 7, 11, and 17 days. Other cultures were treated with either OSM + TNF or vehicle for 7, 11, and 17 days followed by stimulation with either IGF or vehicle control for 14 days. Subsequently, cartilage explants were paraffin-embedded and stained for aggrecan content as described in Materials and methods. As a control experiment, articular cartilage explants were cultured for 21 days with vehicle, OSM + TNF, IGF, or metabolically inactive (MI) control for 21 days as controls (lower panel). W/O, without stimulation.<p><b>Copyright information:</b></p><p>Taken from "Cartilage degradation is fully reversible in the presence of aggrecanase but not matrix metalloproteinase activity"</p><p>http://arthritis-research.com/content/10/3/R63</p><p>Arthritis Research & Therapy 2008;10(3):R63-R63.</p><p>Published online 30 May 2008</p><p>PMCID:PMC2483454.</p><p></p

Cartilage degradation is fully reversible in the presence of aggrecanase but not matrix metalloproteinase activity-0

II synthesis in cartilage explants was measured by the concentration of N-terminal pro-peptides of type II collagen in the conditioned medium using the PIINP enzyme-linked immunosorbent assay (ELISA). The curves represent the release found at the specific day, where the conditioned medium was fully replaced, and the values were accumulated over the entire period. Vehicle control, metabolically inactive (MI), O + T, oncostatin M plus tumour necrosis factor. Quantification of collagen type II formation 2 weeks after the catabolic induction. The collagen type II synthesis in the identical 14-day periods with or without IGF stimulation following the three different periods of catabolic stimulation was measured by the PIINP ELISA. The conditioned medium was fully replaced three times a week. The results show the accumulated release of collagen type II pro-peptide during the two weeks with anabolic stimulation (insulin growth factor, IGF) and without stimulation (vehicle). IGF-I significantly induced collagen type II formation at low and intermediate catabolic insult, but not at maximal insult. *< 0.05. PIINP, N-terminal pro-peptide of pro-collagen type II.<p><b>Copyright information:</b></p><p>Taken from "Cartilage degradation is fully reversible in the presence of aggrecanase but not matrix metalloproteinase activity"</p><p>http://arthritis-research.com/content/10/3/R63</p><p>Arthritis Research & Therapy 2008;10(3):R63-R63.</p><p>Published online 30 May 2008</p><p>PMCID:PMC2483454.</p><p></p

Patients with rheumatoid arthritis have an altered circulatory aggrecan profile-5

, or treated with pro-inflammatory cytokines 10 ng/ml oncostatin M (OSM) in combination with 20 ng/ml tumour necrosis factor alpha (TNFα) (-◆-). The conditioned medium from four independent wells was measured for the presence of aggrecanase-generated ARGSVI-G2 fragments at each collected time-point , or accumulated throughout the study-period. As negative control, explants were frozen and thawed four times in liquid nitrogen (--). The asterisks indicate significant differences (P < 0.05). For the statistical analysis, two-tailed non-parametric t tests were used.<p><b>Copyright information:</b></p><p>Taken from "Patients with rheumatoid arthritis have an altered circulatory aggrecan profile"</p><p>http://www.biomedcentral.com/1471-2474/9/74</p><p>BMC Musculoskeletal Disorders 2008;9():74-74.</p><p>Published online 28 May 2008</p><p>PMCID:PMC2426686.</p><p></p

Patients with rheumatoid arthritis have an altered circulatory aggrecan profile-2

Thritis (RA) (N = 38) were analyzed for their anti-CCP level. The activities are log-transformed data and the values are mean + SEM. The asterisks indicate significant differences (P < 0.05). For the statistical analysis, two-tailed non-parametric t-tests were used.<p><b>Copyright information:</b></p><p>Taken from "Patients with rheumatoid arthritis have an altered circulatory aggrecan profile"</p><p>http://www.biomedcentral.com/1471-2474/9/74</p><p>BMC Musculoskeletal Disorders 2008;9():74-74.</p><p>Published online 28 May 2008</p><p>PMCID:PMC2426686.</p><p></p

Patients with rheumatoid arthritis have an altered circulatory aggrecan profile-4

From patients with Rheumatoid arthritis (RA) (lane 3) and the membrane was stained with the monoclonal antibody F-78 raised against intact bovine aggrecan, binding to G1 and G2, or with BC-3 raised against the aggrecanase-generated ARGSVI sequence. As control, plasma samples from 22 healthy individuals were used (lane 2). A standard molecular weight marker was also run to determine the size of the detected fragments (lane 1).<p><b>Copyright information:</b></p><p>Taken from "Patients with rheumatoid arthritis have an altered circulatory aggrecan profile"</p><p>http://www.biomedcentral.com/1471-2474/9/74</p><p>BMC Musculoskeletal Disorders 2008;9():74-74.</p><p>Published online 28 May 2008</p><p>PMCID:PMC2426686.</p><p></p

Patients with rheumatoid arthritis have an altered circulatory aggrecan profile-6

, or treated with pro-inflammatory cytokines 10 ng/ml oncostatin M (OSM) in combination with 20 ng/ml tumour necrosis factor alpha (TNFα) (-◆-). The conditioned medium from four independent wells was measured for the presence of G1/G2 molecules at each collected time-point , or accumulated throughout the study-period. As negative control, explants were frozen and thawed four times in liquid nitrogen (--). The asterisks indicate significant differences (P < 0.05). For the statistical analysis, two-tailed non-parametric t tests were used.<p><b>Copyright information:</b></p><p>Taken from "Patients with rheumatoid arthritis have an altered circulatory aggrecan profile"</p><p>http://www.biomedcentral.com/1471-2474/9/74</p><p>BMC Musculoskeletal Disorders 2008;9():74-74.</p><p>Published online 28 May 2008</p><p>PMCID:PMC2426686.</p><p></p

Patients with rheumatoid arthritis have an altered circulatory aggrecan profile-3

Thritis (RA) (N = 38) were analyzed in the G1/G2 assay. The concentrations are log-transformed data and the values are mean + SEM. The asterisks indicate significant differences (P < 0.05). For the statistical analysis, two-tailed non-parametric t-tests were used.<p><b>Copyright information:</b></p><p>Taken from "Patients with rheumatoid arthritis have an altered circulatory aggrecan profile"</p><p>http://www.biomedcentral.com/1471-2474/9/74</p><p>BMC Musculoskeletal Disorders 2008;9():74-74.</p><p>Published online 28 May 2008</p><p>PMCID:PMC2426686.</p><p></p