Extracellular Matrix

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Hydroxytyrosol targets extracellular matrix remodeling by endothelial cells and inhibits both ex vivo and in vivo angiogenesis

Hydroxytyrosol targets extracellular matrix remodeling by endothelial cells and inhibits both ex vivo and in vivo angiogenesis

bioactive compounds, such as hydroxytyrosol. Previously, we demonstrated that hydroxytyrosol inhibits angiogenesis in vitro. The present study aimed to: i) get further insight into the effects of hydroxytyrosol on extracellular matrix remodeling; and ii) test whether hydroxytyrosol is able to inhibit angiogenesis ex vivo and in vivo. Hydroxytyrosol induced a shift toward inhibition of proteolysis in endothelial cells, with decreased expression of extracellular matrix remodeling-enzyme coding genes and increased levels of some of their inhibitors. Furthermore, this work demonstrated that hydroxytyrosol, at concentrations within the range of its content in virgin olive oil that can be absorbed from moderate and sustained virgin olive oil consumption, is a strong inhibitor of
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21 Lee mas

Functional role of extracellular matrix proteins and their receptors in apoptosis and cell survival

Functional role of extracellular matrix proteins and their receptors in apoptosis and cell survival

All classes of extracellular matrix molecules may now be considered as macromolecules involved in growth control. However, the information network in which the proteoglycans of the extracellular matrix are involved has only recently been intensively studied. There are two classes of proteoglycans, which at present appear of be capable to directly affecting growth. Members of the lectican family of proteoglycans represent one class; the other one is presently represented only by two members of the small proteoglycan family containing leucine-rich motifs in their core proteins. From other reports, it became evident that in some systems, growth-related effects appear to be mediated via the glycosaminoglycan chain regardless of the biological properties of the core proteins themselves. In these cases, the abundance of a given saccharide structure at the place of action is obviously of greater importance than the fine- tuning of the expression of the proteoglycan itself. It should also be stressed that in order to understand a proteoglycan's role in growth control, it is certainly an oversimplification to only consider the interaction with a cell surface receptor. The temporal and spatial regulation of proteoglycan expression and the possible interactions with other molecules, such as cell adhesion molecules, have to be taken into account as well. Hence, the multifaceted extracellular milieu created by the matrix macromolecules together with the multitude of soluble growth mediators, which are contained within this matrix, yield a level of complexity that cannot be traced back to the interaction of a proteoglycan with a signalling membrane receptor alone. It is therefore not surprising that divergent results may be obtained when correlations between proteoglycans and growth are made in a different biological context (Bandtlow and Zimmermann, 2000).
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174 Lee mas

Inhibition of extracellular matrix assembly induces the expression of osteogenic markers in skeletal muscle cells by a BMP 2 independent mechanism

Inhibition of extracellular matrix assembly induces the expression of osteogenic markers in skeletal muscle cells by a BMP 2 independent mechanism

Skeletal muscle cells are a helpful model for studying cell commitment and differentiation. During skeletal muscle development, fusion of mononuclear myoblasts to form multinucleated myotubes is a central event. This process is partially controlled by the sequential expression of some regulatory proteins, the myogenic regulatory transcription factors (MRFs) of the MyoD family (MyoD, Myf-5, myo- genin and MRF4). Forced expression of MRFs in different mesenchymatic cell lines can induce their transdifferenti- ation into skeletal muscle [2,3]. The expression and activ- ity of these master genes are regulated by several polypeptide growth factors as well as by retinoic acid [4- 7]. The presence of extracellular matrix (ECM) is critical for a proper skeletal muscle differentiation. For instance, inhibitors of collagen synthesis have been shown to inhibit myoblast differentiation [8,9]. Addition of either RGDS peptides or antibodies against integrin receptor to myoblast cultures has also a strong inhibitory effect on muscle differentiation [10,11]. We have shown that inhibitors of proteoglycan synthesis, such as sodium chlo- rate and β-D-xylosides, produce a strong inhibition of ECM assembly that is followed by repression of skeletal muscle differentiation [11,12], even though the MRF myogenin is expressed and properly localized at the nuclei. This inhibition can be totally rescued by the addi- tion of an exogenous ECM, suggesting that the ECM and its receptors provide an appropriate and permissive envi- ronment for lineage-specific cell differentiation [11]. Studies on stem cells transplantation have highlighted the role of local tissue signals for specific cell-type determina- tion, but the relative contribution of intrinsic or genetic signals and extrinsic or ECM signals in cell behavior are not completely understood. Within skeletal muscle tissue specific cells exhibit apparent stem-cell like plasticity [13- 16]. BMP-2 treatment of the mouse myoblast cell line C2C12 [17] and muscle satellite cells isolated from adult mice [18] inhibits myotube formation and induces the expression of alkaline phosphatase activity (ALP) and osteocalcin, changing their differentiation pathway into the osteoblastic lineage. Interestingly, in several muscular diseases [19-21] and animal models for skeletal muscle dystrophy [22], the level of ALP is increased. We have studied microenvironmental changes of skeletal muscle in the mdx mouse, an animal model of Duchenne muscular
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17 Lee mas

Nitric Oxide Induces Cardiac Protection by Preventing Extracellular Matrix Degradation through the Complex Caveolin 3/EMMPRIN in Cardiac Myocytes

Nitric Oxide Induces Cardiac Protection by Preventing Extracellular Matrix Degradation through the Complex Caveolin 3/EMMPRIN in Cardiac Myocytes

Inhibition of Extracellular Matrix degradation by nitric oxide (NO) induces cardiac protection against coronary ischemia/reperfusion (IR). Glycosylation of Extracellular Matrix Metallo- proteinase Inducer (EMMPRIN) stimulates enzymatic activation of matrix metalloprotei- nases (MMPs) in the heart, although the mechanisms leading to EMMPRIN glycosylation are poorly understood. We sought to determine if NO may induce cardiac protection by pre- venting glycosylation of EMMPRIN in a mouse model of IR. Here we found that Caveolin-3 binds to low glycosylated EMMPRIN (LG-EMMPRIN) in cardiac cells and in the hearts of healthy mice, whereas IR disrupted the complex in nitric oxide synthase 2 (NOS2) knockout (KO) mice. By contrast, the binding was partially restored when mice were fed with an NO donor (DEA-NO) in the drinking water, showing a significant reduction on infarct size (NOS2KO: 34.6±5 vs NOS2KO+DEA-NO: 20.7±9), in expression of matrix metalloprotei- nases, and cardiac performance was improved (left ventricular ejection fraction (LVEF). NOS2KO: 31±4 vs NOS2KO+DEA-NO: 46±6). The role of Caveolin-3/EMMPRIN in NO- mediated cardiac protection was further assayed in Caveolin-3 KO mice, showing no signifi- cant improvement on infarct size (Caveolin-3 KO: 34.8±3 vs Caveolin-3 KO+DEA-NO:33.7 ±5), or in the expression of MMPs, suggesting that stabilization of the complex Caveolin-3/ LG-EMMPRIN may play a significant role in the cardioprotective effect of NO against IR.
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14 Lee mas

Extracellular Matrix Proteins Substantiate IL 28B T allele Effect on Histological Outcome of Chronic Hepatitis C

Extracellular Matrix Proteins Substantiate IL 28B T allele Effect on Histological Outcome of Chronic Hepatitis C

Other finding of this study, concerns the demonstra- tion of a strict association between IL-28B rs12979860 T allele and elevation levels of HA, laminin, collagen IV and PIIINP in patients serum. Liver cirrhosis is associated with swathes of dense ECM rich in elastin fibrillar colla- gens and other matrix proteins. Indeed, elevation of these proteins is used as a pathological benchmark for liver dis- ease severity. 19 HA levels increase with the progression of

8 Lee mas

Histological and histochemical evaluation of human oral mucosa constructs developed by tissue engineering

Histological and histochemical evaluation of human oral mucosa constructs developed by tissue engineering

Primary cultures of oral fibroblasts and keratinocytes On arrival at the laboratory, all samples were washed twice in phosphate buffered saline (PBS) and incubated overnight at 37°C in a mixture of DMEM and 2 mg/ml of Clostridium histolyticum collagenase I (Gibco BRL Life Technologies, Karlsruhe, Germany). This enzymatic treatment is able to digest all the extracellular matrix of the oral chorion and release the fibroblasts entrapped there. Once the samples were digested, and to obtain primary cultures of human oral fibroblasts, detached connective tissue components, including fibroblasts, were collected by centrifugation and expanded in culture flasks containing DMEM medium supplemented with antibiotics (100 U/ml of penicillin G, 100 µg/ml of streptomycin, and 0.25 µg/ml of amphotericin B) and 10% of fetal bovine serum (FBS) using standard culture conditions. Then, undigested oral epithelium was washed in PBS, cut into small pieces and co-cultured with a layer of mitomycin C-treated (10 mg/ml) 3T3 feeder cells (8-10x10 3 cell/cm 2 ) (Rheinwald and Green,
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10 Lee mas

Liver fibrosis and inflammation  A review

Liver fibrosis and inflammation A review

As a consequence of chronic tissue damage, Stellate Cells (SC) as well as other extracellular matrix – produc- ing cells such as fibroblasts and myofibroblasts, undergo a process of activation toward a phenotype characterized by increased proliferation, motility, contractility, and syn- thesis of extracellular matrix components. In the liver SC are located in the space of Disse in close contact with hepatocytes and sinusoidal endothelial cells. Hepatic SC express vimentin, desmin,a smooth muscle actin (a-SMA) glial fibrillary acidic protein, nestin, neural cell adhesion molecule and synaptophysin. 28 Quiescent SC are non –
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6 Lee mas

Image analysis in Gomori´s trichrome stain of skeletal muscles subjected to ischemia and reperfusion injury

Image analysis in Gomori´s trichrome stain of skeletal muscles subjected to ischemia and reperfusion injury

Each section of muscle was cut by using a freezing microtome at a temperature of -20ºC before study. Each cut was placed in ionized slides for adequate adherence. Each slide contained six to eight cuts. The quality and correct orientation of tissues were observed by hematoxylin and eosin stain. After that, Gomori’strichrome stain was performed (Table 1). Gomori’strichrome stain is a conventional histochemical technique useful to observe the intramuscular extracellular matrix visible in a green contrast. The muscle fibers appeared red or more intense. The abnormal vesicles in the cytoplasm stain reddish (Sanoudou et al., 2006).
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11 Lee mas

The function of MicroRNA in hepatitis B virus related liver diseases: from Dim to Bright

The function of MicroRNA in hepatitis B virus related liver diseases: from Dim to Bright

MicroRNAs represent a class of non-coding RNA molecules that negatively regulate gene expression either by repressing translation or by inducing degradation of messenger RNA. Studies have shown that, as regula- tors of gene expression, microRNAs are widely involved in various human diseases, including hepatitis B vi- rus-related liver diseases. By modulating hepatitis B virus replication, regulating extracellular matrix formation, as well as silencing tumor suppressor genes, these small molecules are implicated in the devel- opment of chronic hepatitis, liver fibrosis/cirrhosis, and hepatocellular carcinoma caused by hepatitis B vi- rus infection. In addition, current researches indicated a potential role of microRNA as diagnostic markers and therapeutic targets. In conclusion, microRNAs are promising tools in the diagnosis and treatment of hepatitis B virus -related liver diseases.
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7 Lee mas

Título3, 3′, 5 triiodo L thyronine increases in vitro chondrogenesis of mesenchymal stem cells from human umbilical cord stroma through SRC2

Título3, 3′, 5 triiodo L thyronine increases in vitro chondrogenesis of mesenchymal stem cells from human umbilical cord stroma through SRC2

Chondrocyte proliferation and differentiation are regulated by various endocrine, paracrine, and autocrine agents, including growth, thyroid and sex hormones, beta-catenin, bone morphogenetic proteins, insulin-like growth factor, iodothyronine deiodinase, leptin, nitric oxide, transforming growth factor-β, and vitamin D metabolites [Burdan et al., 2009]. Our group directed the differentiation of MSCs from human different tissues like synovial membrane toward chondrocyte-like cells testing different mediums and standard micromass conditions, that is, as pellets [Arufe et al., 2009, 2010], it was demonstrated that spheroid culture of MSC is a valuable method to direct differentiation towards chondrocyte-like cells, however, an optimum amount of aggrecan, the most important proteoglycan in the extracellular matrix of the joint cartilage, was not consistently achieved [Arufe et al., 2011a]. Characterization of MSCs from umbilical cord source by flow cytometry showed in the Figure 1A confirm previous results already published [Arufe et al., 2011b; De la Fuente et al., 2012; Kawata et al., 2012], indicating that our MSCs from human umbilical cord stroma present MSCs markers. Different dosages of T3 and PRL were added to chondrogenic medium to test their effect on chondrogenesis founding PRL had no effect on the chondrogenic process that could be found by qRT-PCR or immunohischemistry analyses. These results were in concordance with results published by Seriwatanachai et al. [2012] who provided evidence that the PRL increased endochondral bone growth and bone elongation, presumably by accelerating apoptosis of hypertrophic chondrocytes in the growth plate and/or subsequent chondrogenic matrix mineralization but not related with chondrogenesis onset. After performing dose-response experiments, qRT-PCR and immunohistochemistry results
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19 Lee mas

TítuloProteome analysis during chondrocyte differentiation in a new chondrogenesis model using human umbilical cord stroma mesenchymal stem cells

TítuloProteome analysis during chondrocyte differentiation in a new chondrogenesis model using human umbilical cord stroma mesenchymal stem cells

Umbilical cord stroma mesenchymal stem cells were differentiated toward chondrocyte-like cells using a new in vitro model that consists of the random formation of spheroids in a medium supplemented with fetal bovine serum on a nonadherent surface. The medium was changed after 2 days to one specific for the induction of chondrocyte differentiation. We assessed this model us- ing reverse transcriptase-polymerase chain reaction, flow cytometry, immunohistochemistry, and secretome analyses. The purpose of this study was to determine which proteins were differentially expressed during chondrogenesis. Differential gel electrophoresis analy- sis was performed, followed by matrix-assisted laser desorption/ionization mass spectrometry protein iden- tification. A total of 97 spots were modulated during the chondrogenesis process, 54 of these spots were identi- fied as 39 different proteins and 15 were isoforms. Of the 39 different proteins identified 15 were down-regulated, 21 were up-regulated, and 3 were up- and down-regu- lated during the chondrogenesis process. Using Path- way Studio 7.0 software, our results showed that the major cell functions modulated during chondrogenesis were cellular differentiation, proliferation, and migra- tion. Five proteins involved in cartilage extracellular ma- trix metabolism found during the differential gel electro- phoresis study were confirmed using Western blot. The results indicate that our in vitro chondrogenesis model is an efficient and rapid technique for obtaining cells similar to chondrocytes that express proteins charac- teristic of the cartilage extracellular matrix. These chon- drocyte-like cells could prove useful for future cell ther-
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10 Lee mas

Mechanisms of angiogenesis in chronic inflammatory liver disease

Mechanisms of angiogenesis in chronic inflammatory liver disease

Endothelial budding is facilitated by vasodilation, loos- ening of interendothelial contacts, and leakiness of preex- isting vessels, which allows extravasation of plasma pro- teins that, together with extracellular matrix components (ECM), lay down a provisional scaffold for migrating endot- helial cells (EC). Nitric oxide (NO), whose angiogenic prop- erties have been characterized, 8 is the main factor responsi-

6 Lee mas

Role of proteoglycans in the regulation of the skeletal muscle fibrotic response

Role of proteoglycans in the regulation of the skeletal muscle fibrotic response

Myogenesis in the embryo and the adult mammal con- sists of a highly organized and regulated sequence of cellular processes aimed at forming or repairing muscle tissue. This sequence includes cell proliferation, migra- tion, and differentiation. Proteoglycans (PGs) play critical roles in skeletal muscle physiology. Structur- ally, they are composed of a core protein to which gly- cosaminoglycan (GAG) chains are covalently attached (chondroitin/dermatan, keratan, and heparan, among others), giving them various and different specific activities. PGs can be found associated with the plasma membrane and the extracellular matrix (ECM). The skeletal muscles express different heparan sulfate PGs (HSPGs) and the small leucine-rich PGs (SLRPs).
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9 Lee mas

Matrix estimation using matrix forgetting factor and instrumental variable for nonstationary sequences with time variant matrix gain

Matrix estimation using matrix forgetting factor and instrumental variable for nonstationary sequences with time variant matrix gain

In this paper, the authors going to describe a good enough estimator considering a system with nonstationary time variant properties with respect to input and output qualities. The techniques used are Instrumental Variable (IV) and Matrix Forgetting Factor (MFF). The results previously obtained by (Poznyak and Medel 1999 a , 1999 b ) were the basis of this paper. The theoretical description

8 Lee mas

Extracellular Vesicles From the Helminth Fasciola hepatica Prevent DSS-Induced Acute Ulcerative Colitis in a T-Lymphocyte Independent Mode

Extracellular Vesicles From the Helminth Fasciola hepatica Prevent DSS-Induced Acute Ulcerative Colitis in a T-Lymphocyte Independent Mode

The complexity of the pathogenesis of inflammatory bowel disease (ulcerative colitis and Crohn’s disease) has led to the quest of empirically drug therapies, combining immunosuppressant agents, biological therapy and modulators of the microbiota. Helminth parasites have been proposed as an alternative treatment of these diseases based on the hygiene hypothesis, but ethical and medical problems arise. Recent reports have proved the utility of parasite materials, mainly excretory/secretory products as therapeutic agents. The identification of extracellular vesicles on those secreted products opens a new field of investigation, since they exert potent immunomodulating effects. To assess the effect of extracellular vesicles produced by helminth parasites to treat ulcerative colitis, we have analyzed whether extracellular vesicles produced by the parasitic helminth Fasciola hepatica can prevent colitis induced by chemical agents in a mouse model. Adult parasites were cultured in vitro and secreted extracellular vesicles were purified and used for immunizing both wild type C57BL/6 and RAG1 − / −
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13 Lee mas

Clinicopathologic features of dual chronic hepatitis B and C infection: A comparison with single hepatitis B, C and delta infections

Clinicopathologic features of dual chronic hepatitis B and C infection: A comparison with single hepatitis B, C and delta infections

Activation of the normal RAS signaling pathway is initiated by the interaction of several cytokines, hor- mones and extracellular growth factors with their ty- rosine-kinases receptors (TKRs). As a result, ligand binding induces receptor dimerization and autophos- phorylation, activating downstream intracellular signal cascades. First, there is recruitment of guanine nucle- otide exchange factors (GEFs), such as RAS-GRF and SOS protein (mammalian homologue of the Drosophila son of sevenless gene product), to the inner surface of the cell membrane where RAS protein is also located af- ter prenylation. RAS is a membrane-bound G protein. The biological activity of RAS is regulated through the
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6 Lee mas

Matrix moment perturbations and the inverse Szegő matrix transformation

Matrix moment perturbations and the inverse Szegő matrix transformation

2.2. Matrix orthogonal polynomials on the unit circle. Let T := {z ∈ C : |z| = 1} and D := {z ∈ C : |z| < 1} be the unit circle and unit disc, respectively, where z ∈ T is parametrized as z = e iθ with θ ∈ (−π, π]. In what follows, σ = (σ i,j ) l i,j=1 will be a Hermitian matrix measure supported on T (on all or part of

21 Lee mas

Glutamatergic signaling in proximal tubular cells maintains the epithelial phenotype and decreases epithelial-mesenchymal transition

Glutamatergic signaling in proximal tubular cells maintains the epithelial phenotype and decreases epithelial-mesenchymal transition

synthesis of mesenchymal markers, such as α -smooth muscle actin ( α -SMA), vimentin and fibroblast specific protein-1 (FSP1) which now defines their new morphology and phenotype. This stage is also associated with the reorganization of the cytoskeleton which provides a structural foundation in defining the morphology of the transformed migratory cell 3,13 . The third step of tubular EMT is characterized by the upregulation of matrix metalloproteinases (MMP-2 and MMP-9) 3,13,21,34 that disrupt the TBM. TBM is the substantial component of renal tubules that promotes diverse cell-matrix interactions crucial for the maintenance of the epithelial phenotype and normal function of tubular epithelial cell 34 . Disruption of TBM is a delayed event that follows the loss of epithelial adhesion and de novo expression of α -SMA and it is of vital importance because it facilitates the subsequent step of tubular EMT - process of invasion and migration of transformed cells toward the interstitial compartments of the kidneys.
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223 Lee mas

Matrix : cuando al comecocos no le quedan opciones

Matrix : cuando al comecocos no le quedan opciones

Los hermanos Wachowski, Larry y Andy, escriben y dirigen Matrix, y la estrenan en el año 1999. Es el momento en el que la sospecha de lo que pueden llegar a ser las tecnologías digitales se vive más intensamente: hemos superado el momento de la navegación y entramos de lleno en el mundo de la inmersión. Es mas, la revolución tecnológica puede llegar a dominar la especie humana. Enseguida nos daremos cuenta de que las cosas no son como parecían, pero esas expectativas hacen que la película de Matrix se estrene en un momento especialmente propenso a la especulación del futuro tecnológico.
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10 Lee mas

Ansoffs Matrix Basic Concepts

Ansoffs Matrix Basic Concepts

The need for this information means that you may find yourself in strategy meetings; a familiarity with the underlying business analysis techniques and jargon can help you to make a valuable contribution by bringing your own area of expertise into the discussion. The Ansoff Matrix, created by the American planning expert Igor Ansoff, is a strategic planning tool that links an organization’s marketing strategy with its general strategic di- rection. It presents four alternative growth strategies in the form of a 2x2 table or matrix. One dimension of the matrix considers ‘products’ (existing and new) and the other di- mension considers ‘markets’ (existing and new).
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28 Lee mas

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