The balance between matrix metalloproteinases (MMPs) and their inhibitors, the tissue inhibitors of (TIMPs), is pivotal in remodeling extracellular matrix. TGF-beta has profound effects on homeostasis, part via its ability to alter this at level gene expression. intracellular signaling pathways by which mediates actions include Smad pathway, specific superfamily, but also, for example, mitogen-activated protein kinase pathways; furthermore, cross-talk Smads other modifies response....

Abstract Objective Previous studies have reported elevated levels of interleukin‐1 (IL‐1) and oncostatin M (OSM) in rheumatoid joints, as well the synergistic degradation human articular cartilage by this cytokine combination. The present study was undertaken to investigate ability IL‐1 OSM modulate gene expression matrix metalloproteinase (MMP), ADAM, ADAM‐TS (ADAM with thrombospondin motifs) family members chondrocytes. Methods T/C28a4 chondrocytes were stimulated for 2–48 hours and/or...

Objectives To investigate the effect of leptin on cartilage destruction. Methods Collagen release was assessed in bovine explant cultures, while collagenolytic and gelatinolytic activities culture supernatants were determined by bioassay gelatin zymography. The expression matrix metalloproteinases (MMP) analysed real-time RT–PCR. Signalling pathway activation studied immunoblotting. Leptin levels cultured osteoarthritic joint infrapatellar fat pad or peri-enthesal deposit measured...

Expression of rat procathepsin B in yeast led to the secretion both latent and mature forms enzyme. Culture presence a cysteine proteinase inhibitor prevented this processing. We have expressed purified mutant form whose active-site residue has been changed serine, which also lacks glycosylation site region protein. This non-active protein was secreted essentially an unprocessed form. The incubated with variety proteinases, results indicate that cathepsins D L, as well cathepsin itself, can...

<h3>Objective</h3> To use a computational approach to investigate the cellular and extracellular matrix changes that occur with age in knee joints of mice. <h3>Methods</h3> Knee from an inbred C57/BL1/6 (ICRFa) mouse colony were harvested at 3–30 months age. Sections stained H&amp;E, Safranin-O, Picro-sirius red antibodies metalloproteinase-13 (MMP-13), nitrotyrosine, LC-3B, Bcl-2, cleaved type II collagen used for immunohistochemistry. Based on this other data literature, computer...

Osteoarthritis is a degenerative joint disease characterized by progressive and irreversible loss of the articular cartilage, due in main part to cleavage type II collagen within matrix enzyme metalloproteinase (MMP)13. Here, we examined methylation status MMP13 promoter report demethylation specific CpG dinucleotides its osteoarthritic compared normal which correlates with increased expression. Of sites examined, –104 was consistently demethylated following treatment human chondrocytes 10μM...

Interstitial collagen types I, II and III are highly resistant to proteolytic attack, due their triple helical structure, but can be cleaved by matrix metalloproteinase (MMP) collagenases at a specific site, approximately three‐quarters of the length from N‐terminus each chain. MMP‐2 ‐9 closely related structural level, MMP‐2, not MMP‐9, has been previously described as collagenase. This report investigates ability purified recombinant human MMP‐9 produced in insect cells degrade native I...

The amino acid sequence of stem bromelain, the major cysteine proteinase from pineapple is described. It shows that enzyme a member papain superfamily proteinases, but not very closely related to any other known this group. mutation or deletion several residues have been conserved in proteinases examined previously, including Asn‐175 (papain). We suggest some these changes effect altering active‐site geometry and accounts for resistance inhibition by cystatins E‐64 [L‐3‐carboxy‐2,3‐ trans...

To determine whether other glycoprotein 130 (gp130) binding cytokines can mimic the effects of oncostatin M (OSM) in acting synergistically with interleukin-1alpha (IL-1alpha) to induce cartilage collagen breakdown and collagenase expression, which receptors mediate these effects.The release proteoglycan was assessed bovine human explant cultures. Messenger RNA (mRNA) protein production from immortalized chondrocytes (T/C28a4) analyzed by Northern blotting specific enzyme-linked...

YKL-40 is expressed in arthritic cartilage and produced large amounts by cultured chondrocytes, but its exact role unclear, the identities of physiological ligands remain unknown. Purification from resorbing bovine nasal chondrocyte monolayers demonstrated existence three isoforms, a major minor form third species chondrocytes. Affinity chromatography experiments with purified specific binding all forms to collagen types I, II, III, thus identifying collagens as potential ligands. Binding...

Abstract Objective We have previously reported the up‐regulation of matrix metalloproteinase 10 (MMP‐10) following treatment with procatabolic stimulus interleukin‐1 (IL‐1) and oncostatin M (OSM) in chondrocytes. Although MMP‐10 is closely related to MMP‐3, little known about role cartilage catabolism. The purpose this study was determine whether expressed connective tissue cells assess how it may contribute collagenolysis. Methods MMP gene expression assessed by real‐time polymerase chain...

This study was designed to identify metalloproteinase determinants of macrophage migration and led the specific hypothesis that matrix 10 (MMP10/stromelysin-2) facilitates migration. We first profiled expression all MMPs in LPS-stimulated primary murine bone marrow-derived macrophages Raw264.7 cells found MMP10 stimulated early (3 h) down-regulated later (24 h). Based on this pattern expression, we speculated plays a role responses, such as Indeed, using time lapse microscopy, RNAi silencing...

Proteinase-activated receptor 2 (PAR2) deficiency protects against cartilage degradation in experimental osteoarthritis (OA). The wider impact of this pathway upon OA-associated pathologies such as osteophyte formation and pain is unknown. Herein, we investigated early temporal bone changes OA order to further elucidate the role PAR2 pathogenesis. was induced wild-type (WT) PAR2-deficient (PAR2-/-) mice by destabilisation medial meniscus (DMM). Inflammation, were monitored using histology...

To examine the ability of a broad-spectrum histone deacetylase (HDAC) inhibitor to protect cartilage in vivo, and explore effects class-selective HDAC inhibitors small interfering RNA (siRNA)-induced knockdown HDACs on metalloproteinase expression degradation vitro.A destabilization medial meniscus (DMM) model was used assess vivo activity trichostatin A (TSA). Human articular chondrocytes (HACs) SW-1353 chondrosarcoma cells were treated with cytokines TSA, valproic acid, MS-275, or siRNA,...

Irreversible breakdown of cartilage extracellular matrix (ECM) by the collagenase metalloproteinase 13 (MMP13) represents a key event in osteoarthritis (OA) progression. Although inflammation is most commonly associated with inflammatory joint diseases, it also occurs OA and thus relevant to prevalent tissue destruction. Here, generates cFOS AP-1 early response that indirectly affects MMP13 gene expression. To ascertain more direct effect on prolonged production we examined potential...

Abstract Objective To determine the effects of proinflammatory cytokine combination oncostatin M (OSM) and tumor necrosis factor α (TNFα) on cartilage destruction in both vitro vivo model systems. Methods The release collagen proteoglycan was assessed bovine explant cultures, while messenger RNA (mRNA) from chondrocytes analyzed by Northern blotting. Immunohistochemistry performed sections prepared murine joints following injection adenovirus vectors encoding OSM and/or TNFα. Results + TNFα...

Abstract Objective To identify the genes up‐regulated by interleukin‐1 (IL‐1) in combination with oncostatin M (OSM) chondrocytes that may be involved mechanisms of cartilage repair and degradation. Methods Gene microarray real‐time polymerase chain reaction (PCR) experiments were performed using RNA from SW1353 primary human articular chondrocytes. Sections prepared murine joints, injected adenovirus vectors overexpressing IL‐1 and/or OSM, analyzed immunohistochemistry for selected...

The electronic structure and equilibrium of magnetite $({\text{Fe}}_{3}{\text{O}}_{4})$ in the high temperature cubic $Fd\overline{3}m$ low monoclinic $P2/c$ unit cells have been computed using Perdew-Wang generalized gradient approximation (GGA) to density functional theory (DFT) B3LYP hybrid functional. ground state for GGA-DFT is an itinerant electron metallic cell a charge ordered semiconducting cell. predicted by ${\text{Fe}}_{3}{\text{O}}_{4}$ has calculated with lattice parameters...

Increasing evidence implicates serine proteinases in pathologic tissue turnover. The aim of this study was to assess the role transmembrane proteinase matriptase cartilage destruction osteoarthritis (OA).Serine gene expression femoral head obtained from either patients with hip OA or fracture neck femur (NOF) assessed using a low-density array. effect on collagen breakdown determined degradation models, while matrix metalloproteinase (MMP) analyzed by real-time polymerase chain reaction....

Matrix metalloproteinase-13 (MMP-13) is a uniquely important collagenase that promotes the irreversible destruction of cartilage collagen in osteoarthritis (OA). Collagenase activation key control point for breakdown to occur, yet our understanding proteinases involved this process limited. Neutrophil elastase (NE) well-described proteoglycan-degrading enzyme which historically associated with inflammatory arthritis, but more recent evidence suggests potential role OA. In study, we...

ABSTRACT: The treatment of cartilage with mediators initiates the breakdown proteoglycan followed by collagen. This is accompanied modulation different proteinases and inhibitors that include members MMP family TIMPs. We have evidence a chondrocyte membrane‐associated metalloproteinase cleaves aggrecan. activity rapidly induced after stimulation IL‐1 OSM not inhibited TIMPs‐l ‐2 but synthetic inhibitors. same combination cytokines also upregulates collagenases subsequent release collagen...