ICVS 3Bs PT Government Laboratory

Braga, Portugal

ICVS 3Bs PT Government Laboratory

Braga, Portugal

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Santo V.E.,University of Minho | Santo V.E.,ICVS 3Bs PT Government Laboratory | Gomes M.E.,University of Minho | Gomes M.E.,ICVS 3Bs PT Government Laboratory | And 4 more authors.
Tissue Engineering - Part B: Reviews | Year: 2013

The development of controlled release systems for the regeneration of bone, cartilage, and osteochondral interface is one of the hot topics in the field of tissue engineering and regenerative medicine. However, the majority of the developed systems consider only the release of a single growth factor, which is a limiting step for the success of the therapy. More recent studies have been focused on the design and tailoring of appropriate combinations of bioactive factors to match the desired goals regarding tissue regeneration. In fact, considering the complexity of extracellular matrix and the diversity of growth factors and cytokines involved in each biological response, it is expected that an appropriate combination of bioactive factors could lead to more successful outcomes in tissue regeneration. In this review, the evolution on the development of dual and multiple bioactive factor release systems for bone, cartilage, and osteochondral interface is overviewed, specifically the relevance of parameters such as dosage and spatiotemporal distribution of bioactive factors. A comprehensive collection of studies focused on the delivery of bioactive factors is also presented while highlighting the increasing impact of platelet-rich plasma as an autologous source of multiple growth factors. © 2013, Mary Ann Liebert, Inc.


Borges S.,University of Minho | Borges S.,ICVS 3Bs PT Government Laboratory | Coimbra B.,University of Minho | Coimbra B.,ICVS 3Bs PT Government Laboratory | And 8 more authors.
Neuropsychopharmacology | Year: 2013

Prenatal stress or exposure to elevated levels of glucocorticoids (GCs) can impair specific neurobehavioral circuits leading to alterations in emotional processes later in life. In turn, emotional deficits may interfere with the quality and degree of social interaction. Here, by using a comprehensive behavioral approach in combination with the measurement of ultrasonic vocalizations, we show that in utero GC (iuGC)-exposed animals present increased immobility in the forced swimming test, pronounced anhedonic behavior (both anticipatory and consummatory), and an impairment in social interaction at different life stages. Importantly, we also found that social behavioral expression is highly dependent on the affective status of the partner. A profound reduction in mesolimbic dopaminergic transmission was found in iuGC animals, suggesting a key role for dopamine (DA) in the etiology of the observed behavioral deficits. Confirming this idea, we present evidence that a simple pharmacological approach - acute L-3,4-dihydroxyphenylacetic acid (L-DOPA) oral administration, is able to normalize DA levels in iuGC animals, with a concomitant amelioration of several dimensions of the emotional and social behaviors. Interestingly, L-DOPA effects in control individuals were not so straightforward; suggesting that both hypo- and hyperdopaminergia are detrimental in the context of such complex behaviors.


Menino J.F.,University of Minho | Menino J.F.,ICVS 3Bs PT Government Laboratory | Almeida A.J.,University of Minho | Rodrigues F.,University of Minho
Methods in Molecular Biology | Year: 2012

Paracoccidioides brasiliensis is a thermal dimorphic fungus which in the host environment exhibits a multinucleated and multibudding yeast form. The cellular and molecular mechanisms underlying these phenotypes remain to be clarified, mostly due to the absence of efficient classical genetic and molecular techniques. Here we describe a method for gene expression knockdown in P. brasiliensis by antisense RNA (aRNA) technology taking advantage of an Agrobacterium tumefaciens-mediated transformation (ATMT) system. Together, these techniques represent a reliable toolbox that can be employed for functional genetic analysis of putative virulence factors and morphogenic regulators, aiming to the identification of new potential drug targets. © 2012 Springer Science+Business Media, LLC.


Batalha V.L.,University of Lisbon | Pego J.M.,University of Minho | Pego J.M.,ICVS 3Bs PT Government Laboratory | Fontinha B.M.,University of Lisbon | And 7 more authors.
Molecular Psychiatry | Year: 2013

Maternal separation (MS) is an early life stress model that induces permanent changes in the central nervous system, impairing hippocampal long-term potentiation (LTP) and spatial working memory. There are compelling evidences for a role of hippocampal adenosine A2A receptors in stress-induced modifications related to cognition, thus opening a potential window for therapeutic intervention. Here, we submitted rats to MS and evaluated the long-lasting molecular, electrophysiological and behavioral impairments in adulthood. We then assessed the therapeutic potential of KW6002, a blocker of A2A receptors, in stress-impaired animals. We report that the blockade of A2A receptors was efficient in reverting the behavior, electrophysiological and morphological impairments induced by MS. In addition, this effect is associated with restoration of the hypothalamic-pituitary-adrenal axis (HPA-axis) activity, as both the plasma corticosterone levels and hippocampal glucocorticoid receptor expression pattern returned to physiological-like status after the treatment. These results reveal the involvement of A2A receptors in the stress-associated impairments and directly in the stress response system by showing that the dysfunction of the HPA-axis as well as the long-lasting synaptic and behavioral effects of MS can be reverted by targeting adenosine A2A receptors. These findings provide a novel evidence for the use of adenosine A2A receptor antagonists as potential therapy against psychopathologies. © 2013 Macmillan Publishers Limited.


Tavares-Valente D.,Institute Investigacao e Formacao Avancada em Ciencias e Tecnologias da Saude | Baltazar F.,University of Minho | Baltazar F.,ICVS 3Bs PT Government Laboratory | Moreira R.,Institute Investigacao e Formacao Avancada em Ciencias e Tecnologias da Saude | And 3 more authors.
Journal of Bioenergetics and Biomembranes | Year: 2013

The multidrug resistance (MDR) phenotype, frequently observed during cancer treatment, is often associated with drug efflux pump activity. However, many other factors are also known to be involved. Cancer cells often rely on aerobic glycolysis for energy production; this is known as the "Warburg effect" and is used as a survival mechanism. Associated to this event, a reverse pH gradient across the cell membrane occurs, leading to cytosol alkalinization and extracellular acidification. In the present study, we investigated the role of different mechanisms involved in MDR, such as altered tumor microenvironment and energetic metabolism. The breast cancer cell line MCF-7, used as model, was exposed to two widely used antitumor drugs, paclitaxel (antimitotic agent) and doxorubicin (alkylating agent). Cancer pH regulation was shown to be crucial for malignant characteristics such as cell migration and drug resistance. Our results showed that a lower extracellular pH induced a higher migratory capacity and higher resistance to the studied chemotherapeutical compounds in MCF-7 cells. Besides the influence of the extracellular pH, the role of the tumor metabolism in the MDR phenotype was also investigated. Pre-treatment with different bioenergetic modulators led to cell ATP depletion and altered lactic acid production and glucose consumption, resulting in increased sensitivity to paclitaxel and doxorubicin. Overall, this study supports the potential use of compounds targeting cell metabolism and tumor microenvironment factors such as pH, as co-adjuvants in conventional chemotherapy. © 2013 Springer Science+Business Media New York.


Luz G.M.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Luz G.M.,ICVS 3Bs PT Government Laboratory | Boesel L.,Max Planck Institute for Polymer Research | Campo A.D.,Max Planck Institute for Polymer Research | And 2 more authors.
Langmuir | Year: 2012

Bioactive glass nanoparticles (BG-NPs) capable of inducing apatite precipitation upon immersion in simulated body fluid (SBF) were patterned on free-standing chitosan membranes by microcontact printing using a poly(dimethylsiloxane) (PDMS) stamp inked in a BG-NPs pad. Formation of the patterns was characterized by scanning electron microscopy (SEM). Mineralization of the bioactive glass patterns was induced in vitro by soaking the samples in SBF over different time points up to 7 days. The confined apatite deposition in the patterned regions with diameters of 50 μm was confirmed by Fourier-transformed infrared spectroscopy (FTIR), energy-dispersive X-ray (EDX) analysis, and SEM. In vitro tests confirmed the preferential attachment and proliferation of L929 cells to the areas printed with BG-NPs of the membranes. This approach permits one to spatially control the properties of biomaterials at the microlevel and could be potentially used in guided tissue regeneration for skin, vascular, articular, and bone tissue engineering and in cellular cocultures or to develop substrates able to confine cells in regions with controlled geometry at the cell's length scale. © 2012 American Chemical Society.


Santo V.E.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Santo V.E.,ICVS 3Bs PT Government Laboratory | Gomes M.E.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Gomes M.E.,ICVS 3Bs PT Government Laboratory | And 4 more authors.
Tissue Engineering - Part B: Reviews | Year: 2013

The potential of growth factors to stimulate tissue healing through the enhancement of cell proliferation, migration, and differentiation is undeniable. However, critical parameters on the design of adequate carriers, such as uncontrolled spatiotemporal presence of bioactive factors, inadequate release profiles, and supraphysiological dosages of growth factors, have impaired the translation of these systems onto clinical practice. This review describes the healing cascades for bone, cartilage, and osteochondral interface, highlighting the role of specific growth factors for triggering the reactions leading to tissue regeneration. Critical criteria on the design of carriers for controlled release of bioactive factors are also reported, focusing on the need to provide a spatiotemporal control over the delivery and presentation of these molecules. © 2013, Mary Ann Liebert, Inc.


Rodrigues A.I.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Gomes M.E.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Leonor I.B.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Leonor I.B.,ICVS 3Bs PT Government Laboratory | And 2 more authors.
Acta Biomaterialia | Year: 2012

Silicon is known to have an influence on calcium phosphate deposition and on the differentiation of bone precursor cells. This study explores the effect of the incorporation of silanol (Si-OH) groups into polymeric scaffolds on the osteogenic differentiation of human adipose stem cells (hASC) cultured under dynamic and static conditions. A blend of corn starch with polycaprolactone (30/70 wt.%, SPCL) was used to produce three-dimensional fibre meshes scaffolds by the wet-spinning technique, and a calcium silicate solution was used as a non-solvent to develop an in situ functionalization with Si-OH groups. In vitro assessment, using hASC, of functionalized and non-functionalized scaffolds was evaluated in either α-MEM or osteogenic medium under static and dynamic conditions (provided by a flow perfusion bioreactor). The functionalized materials, SPCL-Si, exhibit the capacity to sustain cell proliferation and induce their differentiation into the osteogenic lineage. The formation of mineralization nodules was observed in cells cultured on the SPCL-Si materials. Culturing under dynamic conditions using a flow perfusion bioreactor was shown to enhance the hASC proliferation and differentiation and a better distribution of cells within the material. The present work demonstrates the potential of these functionalized materials for future applications in bone tissue engineering. Additionally, these results highlight the simplicity, economic and reliable production process of those materials. © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.


Correia C.R.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Correia C.R.,ICVS 3Bs PT Government Laboratory | Reis R.L.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Reis R.L.,ICVS 3Bs PT Government Laboratory | And 2 more authors.
Biomacromolecules | Year: 2013

Liquified capsules featuring (i) an external shell by layer-by-layer assembly of poly(l-lysine), alginate, and chitosan, and encapsulating (ii) surface functionalized poly(l-lactic acid) (PLLA) microparticles were developed. We hypothesize that, while the liquified environment enhances the diffusion of essential molecules for cell survival, microparticles dispersed in the liquified core of capsules provide the physical support required for cellular functions of anchorage-dependent cells. The influence of the incorporation of PLL on the regime growth, thickness, and stability was analyzed. Results show a more resistant and thicker film with an exponential build-up growth regime. Moreover, capsules ability to support cell survival was assessed. Capsules containing microparticles revealed an enhanced biological outcome in cell metabolic activity and proliferation, suggesting their potential to boost the development of innovative biomaterial designs for bioencapsulation systems and tissue engineering products. © 2013 American Chemical Society.


Duarte A.R.C.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Duarte A.R.C.,ICVS 3Bs PT Government Laboratory | Silva S.S.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Silva S.S.,ICVS 3Bs PT Government Laboratory | And 4 more authors.
Green Chemistry | Year: 2012

In this work, the ability to foam semi-crystalline natural-based polymers by supercritical fluid technology is evaluated. The application of this technique to natural polymers has been limited due to the fact that they are normally semi-crystalline polymers, which do not plasticize in the presence of carbon dioxide. This can be overcome by the use of plasticizers, such as glycerol, which is a commonly used plasticizer, or ionic liquids, which have recently been proposed as plasticizing agents for different polymers. Following the green chemistry principles, the main aim is, hereafter, the design and development of new 3D architectures of natural-based polymers, combining ionic liquids (IL) and supercritical fluid (SCF) technology. A polymeric blend of starch, one of the most abundantly occurring natural polymers, and poly-ε-caprolactone, a synthetic polymer, which is a biodegradable aliphatic polyester commonly used in an array of biomedical applications (SPCL), was processed by supercritical fluid foaming, at different operating conditions, namely pressure (10.0 up to 20.0 MPa), temperature (35 up to 60 °C) and soaking time (30 min up to 3 h). The ionic liquid tested in this work was 1-butyl-3-methylimidazolium acetate ([bmim]Ac). The interactions between SPCL and [bmim]Ac or glycerol were analysed by Fourier transform infrared spectroscopy, differential scanning calorimetry and by mechanical tests, using both tensile and compressive modes. Morphological analysis, porosity, interconnectivity and pore size distribution of the matrixes were evaluated and the morphology was analyzed by scanning electron microscopy and by micro-computed tomography. To our knowledge the use of ionic liquids as foaming agents is reported here for the first time. The results obtained suggest that this approach can further promote the development of composite polymer-IL materials, particularly for catalysis, chromatography, extraction and separation purposes. © The Royal Society of Chemistry 2012.

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