Browsing by Keyword "Molecular Biology"
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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 Surface chemistry and bioactivity of colloidal particles from industrial kraft lignins(2022-11-01) Gordobil, Oihana; Li, Huisi; Izquierdo, Ana Ayerdi; Egizabal, Ainhoa; Sevastyanova, Olena; Sandak, Anna; BiomaterialesThe morphology control of lignin through particle size reduction to nanoscale seems to be a suitable conversion technology to overcome the intrinsic limitations of its native form to develop a wide range of biomaterials with high performance. Colloidal lignin particles (CLPs) in the range of 150–200 nm were synthesised from hardwood and softwood kraft lignins by the solvent shifting method. The initial molecular features of kraft lignins were evaluated in terms of purity, molecular weight distribution, and chemical functionalities. The impact of the lignin source and structure on the morphology, size distribution, and surface chemistry of CLPs was evaluated by particle size analyser, SEM, TEM and 1H NMR. The results evidenced the influence of the botanical origin on the morphology and surface chemistry of particles. Furthermore, the antioxidant properties and cytotoxicity of lignins and corresponding CLPs, towards lung fibroblast cells were compared. CLPs from hardwood kraft lignins exhibited higher antioxidant power against DPPH free radical and a higher cytotoxic effect (IC30 = 67–70 μg/mL) against lung fibroblast when compared to CLPs from softwood kraft lignin (IC30 = ~91 μg/mL). However, the cytotoxicity of these biomaterials was dose-dependent, suggesting their potential application as active ingredients in cosmetic and pharmaceutic products at low concentrations.Item Waste-derived volatile fatty acids as carbon source for added-value fermentation approaches(2021-05-01) Chalima, Angelina; de Castro, Laura Fernandez; Burgstaller, Lukas; Sampaio, Paula; Carolas, Ana Lúcia; Gildemyn, Sylvia; Velghe, Filip; Ferreira, Bruno Sommer; Pais, Celia; Neureiter, Markus; Dietrich, Thomas; Topakas, Evangelos; De Castro, Laura Fernandez; Tecnalia Research & Innovation; Alimentación SostenibleThe establishment of a sustainable circular bioeconomy requires the effective material recycling from biomass and biowaste beyond composting/fertilizer or anaerobic digestion/bioenergy. Recently, volatile fatty acids attracted much attention due to their potential application as carbon source for the microbial production of high added-value products. Their low-cost production from different types of wastes through dark fermentation is a key aspect, which will potentially lead to the sustainable production of fuels, materials or chemicals, while diminishing the waste volume. This article reviews the utilization of a volatile fatty acid platform for the microbial production of polyhydroxyalkanoates, single cell oil and omega-3 fatty acids, giving emphasis on the fermentation challenges for the efficient implementation of the bioprocess and how they were addressed. These challenges were addressed through a research project funded by the European Commission under the Horizon 2020 programme entitled ‘VOLATILE—Biowaste derived volatile fatty acid platform for biopolymers, bioactive compounds and chemical building blocks’.