Browsing by Author "Madarieta, Iratxe"
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Item 3D cell cultures as prospective models to study extracellular vesicles in cancer(2021-01-15) Bordanaba-Florit, Guillermo; Madarieta, Iratxe; Olalde, Beatriz; Falcón-Pérez, Juan M.; Royo, Félix; BiomaterialesThe improvement of culturing techniques to model the environment and physiological conditions surrounding tumors has also been applied to the study of extracellular vesicles (EVs) in cancer research. EVs role is not only limited to cell-to-cell communication in tumor physiology, they are also a promising source of biomarkers, and a tool to deliver drugs and induce antitumoral activity. In the present review, we have addressed the improvements achieved by using 3D culture models to evaluate the role of EVs in tumor progression and the potential applications of EVs in diagnostics and therapeutics. The most employed assays are gel-based spheroids, often utilized to examine the cell invasion rate and angiogenesis markers upon EVs treatment. To study EVs as drug carriers, a more complex multicellular cultures and organoids from cancer stem cell populations have been developed. Such strategies provide a closer response to in vivo physiology observed responses. They are also the best models to understand the complex interactions between different populations of cells and the extracellular matrix, in which tumor-derived EVs modify epithelial or mesenchymal cells to become protumor agents. Finally, the growth of cells in 3D bioreactor-like systems is appointed as the best approach to industrial EVs production, a necessary step toward clinical translation of EVs-based therapy.Item Candida albicans/Macrophage Biointerface on Human and Porcine Decellularized Adipose Matrices(2021-05-17) Cicuéndez, Mónica; Casarrubios, Laura; Feito, María José; Madarieta, Iratxe; Garcia-Urkia, Nerea; Murua, Olatz; Olalde, Beatriz; Briz, Nerea; Diez-Orejas, Rosalía; Portolés, María Teresa; Biomateriales; SGMacrophages, cells effective in sensing, internalizing and killing Candida albicans, are intertwined with the extracellular matrix (ECM) through different signals, which include the release of specific cytokines. Due to the importance of these interactions, the employment of in vitro models mimicking a fungal infection scenario is essential to evaluate the ECM effects on the macrophage response. In this work, we have analyzed the effects of human and porcine decellularized adipose matrices (DAMs), obtained by either enzymatic or organic solvent treatment, on the macrophage/Candida albicans interface. The present study has allowed us to detect differences on the activation of macrophages cultured on either human- or porcine-derived DAMs, evidencing changes in the macrophage actin cytoskeleton, such as distinct F-actin-rich membrane structures to surround the pathogen. The macrophage morphological changes observed on these four DAMs are key to understand the defense capability of these cells against this fungal pathogen. This work has contributed to the knowledge of the influence that the extracellular matrix and its components can exert on macrophage metabolism, immunocompetence and capacity to respond to the microenvironment in a possible infection scenario.Item Confocal and Histological Features After Poly(Ethylene Glycol) Diacrylate Corneal Inlay Implantation(2019-11) Bidaguren, Aritz; Mendicute, Javier; Madarieta, Iratxe; Garagorri, Nerea; Biomateriales; Tecnalia Research & InnovationPurpose: To evaluate the in vivo biocompatibility of photopolymerized poly(ethylene glycol) diacrylate (PEGDA) intrastromal inlays in rabbit corneas. Methods: Sixty-three eyes of 42 New Zealand rabbits were included. Manual intrastromal pockets were dissected in 42 eyes. PEGDA inlays were obtained using a specifically designed photomask and were inserted in the intrastromal pocket of 21 eyes (inlay group); the remaining 21 right eyes did not receive any implant (pocket-only group). Twenty-one eyes with no intervention were used as controls. In vivo confocal microscopy (IVCM) was performed at every visit. After 2 months, rabbits were sacrificed and corneas removed for histological analysis. Results: Corneas remained clear in all but two animals, and five cases of corneal neovascularization were seen (P = 0.2). Inlays remained stable without evidence of lateral or anterior migration, and no other complications were observed. No changes in anterior and posterior keratocyte density (P = 0.3 and P = 0.1, respectively) or endothelial cell density (P = 0.23) were observed between groups during the study time by IVCM. On pathology samples, thinning of the epithelium over the inlay area and epithelial hyperplasia over the edges were observed. A polygonal empty space with no evidence of PEGDA hydrogel within the midstroma was seen in the inlay group. Keratocytes were normal in shape and number in the vicinity of the PEGDA implant area. Conclusions: Photopolymerized PEGDA intrastromal inlays have shown relatively good safety and stability in rabbit corneas. Inlays were biostable in the corneal environment and remained transparent during follow up. Translational Relevance: The investigated PEGDA is promising for the development of biocompatible intrastromal implants.Item Effects of Human and Porcine Adipose Extracellular Matrices Decellularized by Enzymatic or Chemical Methods on Macrophage Polarization and Immunocompetence(2021-04-08) Cicuéndez, Mónica; Casarrubios, Laura; Feito, María José; Madarieta, Iratxe; Garcia-Urkia, Nerea; Murua, Olatz; Olalde, Beatriz; Briz, Nerea; Diez-Orejas, Rosalía; Portolés, María Teresa; Biomateriales; SGThe decellularized extracellular matrix (ECM) obtained from human and porcine adipose tissue (AT) is currently used to prepare regenerative medicine bio-scaffolds. However, the influence of these natural biomaterials on host immune response is not yet deeply understood. Since macrophages play a key role in the inflammation/healing processes due to their high functional plasticity between M1 and M2 phenotypes, the evaluation of their response to decellularized ECM is mandatory. It is also necessary to analyze the immunocompetence of macrophages after contact with decellularized ECM materials to assess their functional role in a possible infection scenario. In this work, we studied the effect of four decellularized adipose matrices (DAMs) obtained from human and porcine AT by enzymatic or chemical methods on macrophage phenotypes and fungal phagocytosis. First, a thorough biochemical characterization of these biomaterials by quantification of remnant DNA, lipids, and proteins was performed, thus indicating the efficiency and reliability of both methods. The proteomic analysis evidenced that some proteins are differentially preserved depending on both the AT origin and the decellularization method employed. After exposure to the four DAMs, specific markers of M1 proinflammatory and M2 anti-inflammatory macrophages were analyzed. Porcine DAMs favor the M2 phenotype, independently of the decellularization method employed. Finally, a sensitive fungal phagocytosis assay allowed us to relate the macrophage phagocytosis capability with specific proteins differentially preserved in certain DAMs. The results obtained in this study highlight the close relationship between the ECM biochemical composition and the macrophage’s functional role.