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Magalhaes J.,Inibic Hospital Universitario Of runa | Crawford A.,University of Sheffield | Hatton P.V.,University of Sheffield | Blanco F.J.,Inibic Hospital Universitario Of runa | Roman J.S.,CIBER ISCIII
Journal of Bioactive and Compatible Polymers | Year: 2013

Poly[2-ethyl(2-pyrrolidone) methacrylate] and hyaluronic acid hydrogels were synthesized via free-radical polymerization of 2-ethyl(2-pyrrolidone) methacrylate, hyaluronic acid and different crosslinkers. The ability of these hydrogels to induce apatite formation by incubating in simulated body fluid was investigated. The effect of hyaluronic acid content, crosslinkers and immersion time on mineralization behaviour and interface properties as well as the metabolic activity of different cultured cells were also determined. The bioactivity of the poly[2-ethyl(2-pyrrolidone) methacrylate] and hyaluronic acid hydrogels along with cell viability data indicated their potential application in bone tissue engineering. © The Author(s) 2013. Source


Rocha B.,Inibic Hospital Universitario Of runa | Cillero-Pastor B.,FOM Institute for Atomic and Molecular Physics | Eijkel G.,FOM Institute for Atomic and Molecular Physics | Bruinen A.L.,FOM Institute for Atomic and Molecular Physics | And 4 more authors.
Proteomics | Year: 2015

Mesenchymal stem cells (MSC) are an interesting alternative for cell-based therapy of cartilage defects attributable to their capacity to differentiate toward chondrocytes in the process termed chondrogenesis. The metabolism of lipids has recently been associated with the modulation of chondrogenesis and also with the development of pathologies related to cartilage degeneration. Information about the distribution and modulation of lipids during chondrogenesis could provide a panel of putative chondrogenic markers. Thus, the discovery of new lipid chondrogenic markers could be highly valuable for improving MSC-based cartilage therapies. In this work, MS imaging was used to characterize the spatial distribution of lipids in human bone marrow MSCs during the first steps of chondrogenic differentiation. The analysis of MSC micromasses at days 2 and 14 of chondrogenesis by MALDI-MSI led to the identification of 20 different lipid species, including fatty acids, sphingolipids, and phospholipids. Phosphocholine, several sphingomyelins, and phosphatidylcholines were found to increase during the undifferentiated chondrogenic stage. A particularly detected lipid profile was verified by TOF secondary ion MS. Using this technology, a higher intensity of phosphocholine-related ions was observed in the peripheral region of the micromasses collected at day 14. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Source


Mateos J.,Inibic Hospital Universitario Of runa | Lourido L.,Inibic Hospital Universitario Of runa | Fernandez-Puente P.,Inibic Hospital Universitario Of runa | Calamia V.,Inibic Hospital Universitario Of runa | And 5 more authors.
Journal of Proteomics | Year: 2012

The purpose of this study was to identify those proteins relatively more abundant in the synovial fluid (SF) of patients suffering from rheumatoid arthritis (RA) and osteoarthritis (OA) using high performance liquid chromatography coupled to mass spectrometry. 20 individual SF samples from each disease were pooled into two groups (RA and OA) to reduce the contribution of extreme individual values. Prior to the proteomic analysis, samples were immunodepleted from the top 20 most abundant plasma proteins, to enrich the lower-abundance protein fractions. Then, they were subjected to protein size fractioning and in-gel digestion, followed by reversed-phase peptide separation in a nano-LC system and subsequent peptide identification by MALDI-TOF/TOF. This strategy led to the identification of 136 different proteins in SF, which is the largest number of SF proteins described up to date by proteomics.A relative quantification of the proteins between RA and OA was carried out by spectral counting analysis. In RA, our results show a greater relative abundance of proteins related to complement activation, inflammation and the immune response, such as the major matrix metalloproteinases and several neutrophil-related proteins. In OA, we detected an increase in proteins involved in the formation and remodeling of the extracellular matrix (ECM), such as fibronectin, kininogen-1, cartilage acidic protein 1 and cartilage oligomeric matrix protein. The results obtained for MMP-1, BGH3, fibronectin and gelsolin were verified by immunoblotting analyses. Some of the novel proteins identified in this work might be relevant not only for increasing knowledge on the etiopathogenesis of RA and OA processes, but also as putative disease biomarkers, as their presence in SF is a prior step to their dilution in serum. This article is part of a Special Issue entitled: Proteomics: The clinical link. © 2012 Elsevier B.V.. Source


Rocha B.,Inibic Hospital Universitario Of runa | Calamia V.,Inibic Hospital Universitario Of runa | Casas V.,Institute Investigaciones Biomedicas Of Barcelona | Carrascal M.,Institute Investigaciones Biomedicas Of Barcelona | And 3 more authors.
Journal of Proteome Research | Year: 2014

Human mesenchymal stem cells (hMSCs) can be triggered to differentiate toward chondrocytes and thus harbor great therapeutic potential for the repair of cartilage defects in osteoarthritis (OA) and other articular diseases. However, the molecular mechanisms underlying the chondrogenesis process are still in part unknown. In this work, we followed a double-stable isotope labeling by amino acids in cell culture (SILAC) strategy to evaluate the quantitative modulation of the secretome of stem cells isolated from bone marrow (hBMSCs) during the first steps of their chondrogenic differentiation. Analysis by LC-ESI-MS/MS led to the identification of 221 proteins with a reported extracellular localization. Most of them were characteristic of cartilage extracellular matrix, and 34 showed statistically significant quantitative alterations during chondrogenesis. These include, among others, cartilage markers such as Proteoglycan 4 or COMP, anticatabolic markers (TIMP1), reported markers of cartilage development (Versican), and a suggested marker of chondrogenesis, CRAC1. Altogether, our work demonstrates the usefulness of secretome analysis for understanding the mechanisms responsible for cartilage matrix formation, and it reports a panel of extracellular markers potentially useful for the evaluation of tissue development in cell therapy- or tissue engineering-based approaches for cartilage repair. © 2014 American Chemical Society. Source


Ruiz-Romero C.,Inibic Hospital Universitario Of runa | Ruiz-Romero C.,CIBER ISCIII | Calamia V.,Inibic Hospital Universitario Of runa | Albar J.P.,CSIC - National Center for Biotechnology | And 7 more authors.
Journal of Proteomics | Year: 2015

The Spanish Chromosome 16 consortium is integrated in the global initiative Human Proteome Project, which aims to develop an entire map of the proteins encoded following a gene-centric strategy (C-HPP) in order to make progress in the understanding of human biology in health and disease (B/D-HPP). Chromosome 16 contains many genes encoding proteins involved in the development of a broad range of diseases, which have a significant impact on the health care system. The Spanish HPP consortium has developed a B/D platform with five programs focused on selected medical areas: cancer, obesity, cardiovascular, infectious and rheumatic diseases. Each of these areas has a clinical leader associated to a proteomic investigator with the responsibility to get a comprehensive understanding of the proteins encoded by Chromosome 16 genes. Proteomics strategies have enabled great advances in the area of rheumatic diseases, particularly in osteoarthritis, with studies performed on joint cells, tissues and fluids. Biological significance: In this manuscript we describe how the Spanish HPP-16 consortium has developed a B/D platform with five programs focused on selected medical areas: cancer, obesity, cardiovascular, infectious and rheumatic diseases. Each of these areas has a clinical leader associated to a proteomic investigator with the responsibility to get a comprehensive understanding of the proteins encoded by Chromosome 16 genes. We show how the Proteomic strategy has enabled great advances in the area of rheumatic diseases, particularly in osteoarthritis, with studies performed on joint cells, tissues and fluids. This article is part of a Special Issue entitled: HUPO 2014. © 2015 Elsevier B.V. Source

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