MicroRNAs (miRNA) are short RNA molecules regulating the expression of specific mRNAs. We investigated pattern and potential targets mouse miR‐140 found that is specifically expressed in cartilage tissues embryos during both long flat bone development. MiR‐140 was detected limbs E11.5 primorida future bones fore hindlimb across autopod, zeugopod stylopod. All digits E14.5 fore‐ hindlimbs showed accumulation miR‐140, except first digit hindlimb. also cartilagenous base E17.5 skulls sternum,...

To profile the expression of all known members matrix metalloproteinase (MMP), ADAMTS, and tissue inhibitor metalloproteinases (TIMP) gene families in normal cartilage from patients with osteoarthritis (OA).Human was obtained femoral heads at joint replacement for OA or following fracture to neck. Total RNA purified, assayed using quantitative real-time polymerase chain reaction.Several above were regulated OA. Genes that showed increased MMP13, MMP28, ADAMTS16 (all P < 0.001), MMP9, MMP16,...

To use an in vitro model of chondrogenesis to identify microRNAs (miRNAs) with a functional role cartilage homeostasis.The expression miRNAs was measured the ATDC5 cell using microarray and verified quantitative reverse transcription-polymerase chain reaction. MicroRNA localized by situ hybridization. Predicted miRNA target genes were validated 3'-untranslated region-Luc reporter plasmids containing either wild-type sequences or mutants sequence. Signaling through Smad pathway (CAGA)(12)...

Summary Replicative senescence occurs when normal somatic cells stop dividing. Senescent remain viable, but show alterations in phenotype, e.g. altered expression of matrix metalloproteinases ( MMPs ); these enzymes are known to be involved cartilage destruction. It is assumed that deplete their replicative potential during aging, and age a major risk factor for osteoarthritis (OA). Therefore, we hypothesized chondrocytes aging or diseased become senescent with associated phenotypic changes...

Abstract Objective To profile the messenger RNA (mRNA) expression for 23 known genes of matrix metalloproteinases (MMPs), 19 ADAMTS, 4 tissue inhibitors (TIMPs), and ADAM 8, 10, 12, 17 in normal, painful, ruptured Achilles tendons. Methods Tendon samples were obtained from cadavers or patients undergoing surgical procedures to treat chronic painful tendinopathy tendon. Total was extracted mRNA analyzed by quantitative real‐time reverse transcription–polymerase chain reaction, normalized 18S...

<h3>Background</h3> Oxidative stress is proposed as an important factor in osteoarthritis (OA). <h3>Objective</h3> To investigate the expression of three superoxide dismutase (SOD) antioxidant enzymes OA. <h3>Methods</h3> SOD was determined by real-time PCR and immunohistochemistry using human femoral head cartilage. SOD2 Dunkin–Hartley guinea pig knee articular cartilage immunohistochemistry. The DNA methylation status promoter bisulphite sequencing. RNA interference used to determine...

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 Cartilage destruction in osteoarthritis (OA) is thought to be mediated by two main enzyme families; the matrix metalloproteinases (MMPs) are responsible for cartilage collagen breakdown, whereas enzymes from 'a disintegrin and metalloproteinase domain with thrombospondin motifs' (ADAMTS) family mediate aggrecan loss. Tissue inhibitors of (TIMPs) regulate activity these enzymes. Although OA might driven chondrocyte, low-grade synovitis reported patients all grades this disease. Our...

On purification, human fibroblast collagenase breaks down into two major forms (Mr22,000 and Mr 27,000) one minor form (Mr 25,000). The most likely mechanism is autolysis, although the presence of contaminating enzymes cannot be excluded. From N-terminal sequencing studies, 22,000-Mr fragment contains active site; differential binding to concanavalin A shows 25,000-Mr a glycosylated fragment. These low-Mr can separated by Zn2+-chelate chromatography. An activity profile this column, combined...

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...

Objective To study the interaction of interleukin-1α (IL-1α) and oncostatin M (OSM) in promoting cartilage collagen destruction. Methods Bovine, porcine, human chondrocytes were studied culture. The levels collagenase (matrix metalloproteinase 1 [MMP-1]) tissue inhibitor metalloproteinases (TIMP-1) measured by bioassay enzyme-linked immunosorbent assay (ELISA). OSM rheumatoid synovial fluid ELISA. Results When combined with OSM, IL-1α IL-1β, tumor necrosis factor α released proteoglycan from...

Increased collagenase activity in colorectal carcinomas has recently been shown to be associated with increased malignant potential. To determine the tissue distribution of and its specific inhibitor, inhibitor metalloproteinases (TIMP), we carried out an immunohistochemical study on (n = 20), adenomas 7) normal mucosa 6). We found staining for connective stroma carcinomas, as compared mucosa. Little evidence epithelial cell was seen any tissue. In both stromal fibroblasts collagen fibres...

Abstract Cartilage destruction in the arthritides is thought to be mediated by two main enzyme families: matrix metalloproteinases (MMPs) are responsible for cartilage collagen breakdown, and enzymes from ADAMTS (a disintegrin metalloproteinase domain with thrombospondin motifs) family mediate aggrecan loss. Many genes subject transcriptional control regulated, at least part, modifications chromatin, including acetylation of histones. The aim this study was examine impact histone deacetylase...

MicroRNAs have been shown to function in cartilage development and homeostasis, as well progression of osteoarthritis. The objective the current study was identify microRNAs involved onset or early osteoarthritis characterise their chondrocytes. MicroRNA expression mouse knee joints post-DMM surgery measured over 7 days. Expression miR-29b-3p increased at day 1 regulated opposite direction its potential targets. In a model injury end-stage human OA cartilage, miR-29 family also regulated....

Abstract Aims Cardiac fibrosis is a major cause of heart failure (HF), and mediated by the differentiation cardiac fibroblasts into myofibroblasts. However, limited tools are available to block fibrosis. ADAMTS16 member ADAMTS superfamily extracellular protease enzymes involved in matrix (ECM) degradation remodelling. In this study, we aimed establish as key regulator Methods results Western blot qRT–PCR analyses demonstrated that was significantly up-regulated mice with transverse aortic...

Abstract Objective. To compare the levels of collagenase, tissue inhibitor metalloproteinases (TIMP), and collagenase—TIMP complex in synovial fluid (SF) from patients with rheumatoid arthritis (RA) osteoarthritis (OA). This study aims to clarify existing data previously used enzyme or activity assays performed following separation by gel filtration, using a 1‐step enzyme‐linked immunosorbent assay (ELISA) for each component. Methods. Total free TIMP, were measured newly developed, specific...

mRNA profiling is routinely used to identify microRNA targets, however, this high-throughput technology not suitable for identifying targets regulated only at protein level. Here, we have developed and validated a novel methodology based on computational analysis of promoter sequences combined with microarray experiments reveal transcription factors that are direct the Using approach identified Smad3, key factor in TGFβ signaling pathway, as miR-140 target. We showed suppressed pathway...

Abstract microRNAs (miRNAs) are abundantly expressed in development where they critical determinants of cell differentiation and phenotype. Accordingly miRNAs essential for normal skeletal chondrogenesis particular. However, the question which specific to chondrocyte phenotype has not been fully addressed. Using microarray analysis miRNA expression during mesenchymal stem chondrogenic detailed examination role factors, such as SOX9, TGF-β, condensation phase, we characterize repertoire...

To identify osteoarthritis (OA) relevant genes and pathways in damaged undamaged cartilage isolated from the knees of patients with anteromedial gonarthrosis (AMG) - a specific form knee OA.Cartilage was obtained nine undergoing unicompartmental replacement (UKR) for AMG. AMG provides spatial representation OA progression; showing reproducible histologically validated pattern destruction such that within same can be consistently examined. Gene expression analysed by microarray using...