Browsing by Author "Olalde, Beatriz"
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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 Decellularization of xenografted tumors provides cell-specific in vitro 3D environment(2022-08-18) Iazzolino, Gaia; Mendibil, Unai; Arnaiz, Blanca; Ruiz-de-Angulo, Ane; Azkargorta, Mikel; Uribe, Kepa B.; Khatami, Neda; Elortza, Felix; Olalde, Beatriz; Gomez-Vallejo, Vanessa; Llop, Jordi; Abarrategi, Ander; BiomaterialesIn vitro cell culture studies are common in the cancer research field, and reliable biomimetic 3D models are needed to ensure physiological relevance. In this manuscript, we hypothesized that decellularized xenograft tumors can serve as an optimal 3D substrate to generate a top-down approach for in vitro tumor modeling. Multiple tumor cell lines were xenografted and the formed solid tumors were recovered for their decellularization by several techniques and further characterization by histology and proteomics techniques. Selected decellularized tumor xenograft samples were seeded with the HCC1806 human triple-negative breast cancer (TNBC) basal-like subtype cell line, and cell behavior was compared among them and with other control 2D and 3D cell culture methods. A soft treatment using Freeze-EDTA-DNAse allows proper decellularization of xenografted tumor samples. Interestingly, proteomic data show that samples decellularized from TNBC basal-like subtype xenograft models had different extracellular matrix (ECM) compositions compared to the rest of the xenograft tumors tested. The in vitro recellularization of decellularized ECM (dECM) yields tumor-type–specific cell behavior in the TNBC context. Data show that dECM derived from xenograft tumors is a feasible substrate for reseeding purposes, thereby promoting tumor-type–specific cell behavior. These data serve as a proof-of-concept for further potential generation of patient-specific in vitro research models.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.Item Enhancing Osteoconduction of PLLA-Based Nanocomposite Scaffolds for Bone Regeneration Using Different Biomimetic Signals to MSCs(2012-02) Ciapetti, Gabriela; Granchi, Donatella; Devescovi, Valentina; Baglio, Serena R.; Leonardi, Elisa; Martini, Desirèe; Jurado, Maria Jesus; Olalde, Beatriz; Armentano, Ilaria; Kenny, Josè M.; Walboomers, Frank X.; Alava, J.I.; Baldini, Nicola; Tecnalia Research & Innovation; BiomaterialesIn bone engineering, the adhesion, proliferation and differentiation of mesenchymal stromal cells rely on signaling from chemico-physical structure of the substrate, therefore prompting the design of mimetic “extracellular matrix”-like scaffolds. In this study, three-dimensional porous poly-L-lactic acid (PLLA)-based scaffolds have been mixed with different components, including single walled carbon nanotubes (CNT), micro-hydroxyapatite particles (HA), and BMP2, and treated with plasma (PT), to obtain four different nanocomposites: PLLA + CNT, PLLA + CNTHA, PLLA + CNT + HA + BMP2 and PLLA + CNT + HA + PT. Adult bone marrow mesenchymal stromal cells (MSCs) were derived from the femur of orthopaedic patients, seeded on the scaffolds and cultured under osteogenic induction up to differentiation and mineralization. The release of specific metabolites and temporal gene expression profiles of marrow-derived osteoprogenitors were analyzed at definite time points, relevant to in vitro culture as well asin vivo differentiation. As a result, the role of the different biomimetic components added to the PLLA matrix was deciphered, with BMP2-added scaffolds showing the highest biomimetic activity on cells differentiating to mature osteoblasts. The modification of a polymeric scaffold with reinforcing components which also work as biomimetic cues for cells can effectively direct osteoprogenitor cells differentiation, so as to shorten the time required for mineralization.Item Fabrication and Characterization of a Novel Bioactive Poly(ether-ether-ketone) Scaffold for Bone Tissue Engineering (Poster)(MARY ANN LIEBERT, INC, 140 HUGUENOT STREET, 3RD FL, NEW ROCHELLE, NY 10801 USA, 2015-09) Olalde, Beatriz; Oyarbide, Joseba; Ayerdi, Ana; Azpiroz, Patxi; Fernandez, Ruben; Aizpurua, J.; Braceras, I.; Alvarez, Noelia; Briz, N.; Morin, F.O.It is widely accepted that the initial interactions between the cells and implant surface are crucial to clinical success. One of the promising alternative materials for tissue engineering is polyetheretherketone (PEEK) which has good chemical resistance and mechanical properties similar to those of human bones [1]. However, its hydrophobic and chemically inert surface limits local bone attachment [2]. Consequently, there have been efforts to enhance PEEK bone-implant integration by incorporating molecules (such as HA) or making it porous [3]. Regarding to the latest, this study aims to demonstrate a process that can greatly enhance the density and interconnectivity of a PEEK porous product (US2012323339) [4]. Moreover, a novel surface modification through wet-chemistry protocol has been developed for RGD and OGP (osteogenic growth peptide) grafting onto porous PEEK scaffolds (US2013172273) [5]. As a result, the production of porous structures manufactured by thermally induced phase separation technique (TIPS), in combination with particulate leaching method, offers the greatest potential to control pore size and porosity whilst achieving porosity higher than 80%. Besides, surface functionalization of porous PEEK scaffolds with RGD/OGP combination is shown to enhance osteoblast adhesion, proliferation and differentiation, according to the in vitro results. In conclusion, we have developed a convenient “click” chemistry procedure to modify PEEK surfaces with RGD and OGP10-14 molecules, while keeping the bulk properties of PEEK as well as its biocompatibility in terms of cytotoxicity. The improvement of the osteogenic activity of this novel material shows that can be appropriated for bone tissue engineering applications.