Western-blotting detection of osteomodulin using goat polyclonal antisera raised against entire osteomodulin.

<p>(A)(B) rhOMD: human recombinant osteomodulin (positive control), DMEM: concentration unconditioned culture supernatant (negative control), NSC: non sclerotic osteoblasts supernatant (N36, N38, N40), SC: sclerotic osteoblasts supernatant (S36, S38, S40), C1-6: six healthy patients albumin/IgG depleted serum, OA1-6: six OA patients before total knee replacement surgery albumin/IgG depleted serum. (C) Quantification of osteomodulin western blot performed on osteoblasts culture supernatants and on albumin/IgG depleted serum of 6 healthy patients (controls) and 6 patients before total knee replacement (OA). Quantification was made using ImageJ on total band and normalized with total protein charged visualized with reverse total protein stain (Pierce).</p

MS/MS proteomic analysis of osteoblasts secretome.

<p>MS/MS proteomic analysis of osteoblasts secretome.</p

Compounds effects, separately or in combination, on IL-1β stimulated bovine chondrocytes in monolayer (expression).

<p>Results were expressed as percent of IL-1β treated chondrocytes culture (control). *p<0.05 versus control, **p<0.01 versus control, ***p<0.001 versus control, #p<0.05 versus COT, ##p<0.01 versus COT, ###p<0.001 versus COT. (a) IL-6 = interleukin-6, COX2 = cyclooxygenase2, iNOS = inducible nitric oxide synthase, (b) MMP-3 = matrix metalloproteinase-3, ADAMTS4 = a disintegrin and metalloproteinase with thrombospondin type 1 motifs 4 and ADAMTS5 = a disintegrin and metalloproteinase with thrombospondin type 1 motifs 5.</p

Sequences of forward and reverse bovine primers used for PCR.

<p>HPRT = hypoxanthine phosphoribosyltransferase, IL-6 = interleukin-6, COX2 = cyclooxygenase2, iNOS = inducible nitric oxide synthase, MMP-3 = matrix metalloproteinase-3, ADAMTS4 = a disintegrin and metalloproteinase with thrombospondin type I motifs 4, ADAMTS5 = a disintegrin and metalloproteinase with thrombospondin type I motifs 5.</p><p>Sequences of forward and reverse bovine primers used for PCR.</p

OMD1 and OMD2 levels found in healthy and OA human sera.

<p>(A) OMD1, (B) OMD2. Data are presented as scatter dot plots with median from 22 patients in each group. Comparisons of values were performed by Mann Whitney test. (C) Spearman correlation between OMD1 and OMD2 in serum, n = 44.</p

Compounds effects, separately or in combination, on IL-1β stimulated human chondrocytes in monolayer (production).

<p>Results were expressed as percent of IL-1β treated chondrocytes culture (control). *p<0.05 versus control, **p<0.01 versus control, ***p<0.001 versus control, #p<0.05 versus COT, ##p<0.01 versus COT, ###p<0.001 versus COT. (a) NO = nitric oxide, (b) IL-6 = interleukin-6, (c) MMP-3 = matrix metalloproteinase-3.</p

OMD1 and OMD2 levels found in osteoblasts cell culture supernatant.

<p>Data are means +/- SEM from 10 patients. Comparisons of mean values were performed by unpaired t-test.</p

Compounds effects, separately or in combination, on human chondrocytes in monolayer (production).

<p>Results were expressed as percent of control. *p<0.05 versus control, ***p<0.001 versus control, #p<0.05 versus COT, ###p<0.001 versus COT. (a) MMP-3 = matrix metalloproteinase-3, (b, c) aggrecans.</p

Compounds effects on IL-1β induced phosphorylations of NF-κB p65 and IκBα subunits.

<p>Western blot analysis with IL-1β stimulated chondrocytes total protein extracts. Chondrocytes were pre-incubated with C or COT for 24 h and then co-treated with IL-1β (10^-11 M) for 5 or 15 min. The protein extracts were probed for phospho NF-κB p65, phospho IκBα and HSC70 (control) by Western blot analysis using specific antibodies.</p

Compounds effects on IL-1β stimulated bovine chondrocytes in monolayer (expression).

<p>Results were expressed as percent of IL-1β treated chondrocytes culture (control). *p<0.05 versus control, **p<0.01 versus control, ***p<0.001 versus control. (a) IL-6 = interleukin-6, (b) COX2 = cyclooxygenase2, (c) iNOS = inducible nitric oxide synthase, (d) MMP-3 = matrix metalloproteinase-3, (e) ADAMTS4 = a disintegrin and metalloproteinase with thrombospondin type 1 motifs 4 and (f) ADAMTS5 = a disintegrin and metalloproteinase with thrombospondin type 1 motifs 5.</p