European Institute of Excellence on Tissue Engineering and Regenerative Medicine

Guimaraes, Portugal

European Institute of Excellence on Tissue Engineering and Regenerative Medicine

Guimaraes, Portugal
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Song W.,Jilin University | Mano J.F.,University of Minho | Mano J.F.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine
Soft Matter | Year: 2013

Regulation of protein adsorption and cell adhesion on surfaces is a key aspect in the field of biomedicine and tissue engineering. Beside the general studies on hydrophilic/hydrophobic surfaces, there are both fundamental and practical interests to extend the investigation of the interaction between proteins or cells and surfaces to the two extreme wettability ranges, namely superhydrophilicity and superhydrophobicity. This review gave an overview of recent studies on proteins or cells action on these two special wettability ranges. The first part will focus on the interaction between proteins and superhydrophilic/superhydrophobic surfaces. The second part will focus on cells adhesion on these extreme wettable surfaces. Surfaces can be patterned to control in space the wettability within extreme values. As an application of such substrates, flat chips for high-throughput screening are also addressed to offer new insight on the design of a new type of bioanalysis supports. © 2013 The Royal Society of Chemistry.


Borges J.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Borges J.,ICVS 3Bs PT Government Associate Laboratory | Mano J.F.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Mano J.F.,ICVS 3Bs PT Government Associate Laboratory
Chemical Reviews | Year: 2014

This article presents an overview of the different types of intermolecular interactions behind the fabrication of multilayer assemblies using the layer-by-layer (LbL) assembly approach. It comments on the potential impact of each type of intermolecular interaction and materials assembled through them on the development of advanced functional systems or devices for several emerging applications. The discussion begins with a brief overview of the most commonly used bottom-up methods to modify surfaces and fabricate functional multilayer thin films, with a special focus on their main advantages and disadvantages.


Oliveira M.B.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Oliveira M.B.,ICVS 3Bs PT Government Associate Laboratory | Mano J.F.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Mano J.F.,ICVS 3Bs PT Government Associate Laboratory
Trends in Biotechnology | Year: 2014

With the increasing need for biomaterials and tissue engineering alternatives, more accurate, rapid, and cost-saving methods and models to study biomaterial-cell interactions must be developed. We review the evolution of microarray platforms used for such studies in order to meet the criteria of complex tissue engineering biological environments. Particular aspects regarding biomaterials processing, data acquisition, and treatment are addressed. Apart from in vitro array-based strategies, we also address emerging in vivo high-throughput approaches and their associated trends, such as the role of inflammation in regeneration. The up-scaling of high-throughput methods using single cell encapsulation systems is also explored. Possible limitations related to the use of such methods, such as spot-to-spot crosstalk, are also discussed. © 2014 Elsevier Ltd.


Lourenco B.N.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine
Biointerphases | Year: 2012

Surface wettability and topography are recognized as critical factors influencing cell behavior on biomaterials. So far only few works have reported cell responses on surfaces exhibiting extreme wettability in combination with surface topography. The goal of this work is to study whether cell behavior on superhydrophobic surfaces is influenced by surface topography and polymer type. Biomimetic superhydrophobic rough surfaces of polystyrene and poly(L-lactic acid) with different micro/nanotopographies were obtained from smooth surfaces using a simple phase-separation based method. Total protein was quantified and showed a less adsorption of bovine serum albumin onto rough surfaces as compared to smooth surfaces of the same material. The mouse osteoblastic MC3T3-E1 cell line and primary bovine articular chondrocytes were used to study cell attachment and proliferation. Cells attached and proliferate better in the smooth surfaces. The superhydrophobic surfaces allowed cells to adhere but inhibited their proliferation. This study indicates that surface wettability, rather than polymer type or the topography of the superhydrophobic surfaces, is a critical factor in determining cell behavior.


Alves A.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine
Carbohydrate Research | Year: 2010

During the last years, considerable attention has been given to different marine organisms, like algae, as potential sources of valuable materials. The continuous demand for novel materials and technologies is high and research on the underexploited marine green algae, including its polysaccharidic part - ulvan, has increased accordingly. In this research work, a novel method for extraction of ulvan from green algae is proposed and demonstrated successfully. Different characterization techniques were employed to characterize the isolated algal polysaccharide, namely, on what concerns its thermal trace and crystallinity. Upon heating, ulvan behaves as a non-meltable polysaccharide that is thermally stable before degradation at 220 °C. Ulvan is semi-crystalline in nature and possesses high hygroscopic features, as revealed in this research work. Due to its properties, ulvan can be considered, pure or modified, as a versatile biodegradable polymer for different applications, including tissue engineering and regenerative medicine. © 2010 Elsevier Ltd. All rights reserved.


Pina S.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Oliveira J.M.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Reis R.L.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine
Advanced Materials | Year: 2015

Tissue engineering and regenerative medicine has been providing exciting technologies for the development of functional substitutes aimed to repair and regenerate damaged tissues and organs. Inspired by the hierarchical nature of bone, nanostructured biomaterials are gaining a singular attention for tissue engineering, owing their ability to promote cell adhesion and proliferation, and hence new bone growth, compared with conventional microsized materials. Of particular interest are nanocomposites involving biopolymeric matrices and bioactive nanosized fillers. Biodegradability, high mechanical strength, and osteointegration and formation of ligamentous tissue are properties required for such materials. Biopolymers are advantageous due to their similarities with extracellular matrices, specific degradation rates, and good biological performance. By its turn, calcium phosphates possess favorable osteoconductivity, resorbability, and biocompatibility. Herein, an overview on the available natural polymer/calcium phosphate nanocomposite materials, their design, and properties is presented. Scaffolds, hydrogels, and fibers as biomimetic strategies for tissue engineering, and processing methodologies are described. The specific biological properties of the nanocomposites, as well as their interaction with cells, including the use of bioactive molecules, are highlighted. Nanocomposites in vivo studies using animal models are also reviewed and discussed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Silva T.H.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine
Biomatter | Year: 2012

Biomedical field is constantly requesting for new biomaterials, with innovative properties. Natural polymers appear as materials of election for this goal due to their biocompatibility and biodegradability. In particular, materials found in marine environment are of great interest since the chemical and biological diversity found in this environment is almost uncountable and continuously growing with the research in deeper waters. Moreover, there is also a slower risk of these materials to pose illnesses to humans.   In particular, sulfated polysaccharides can be found in marine environment, in different algae species. These polysaccharides don't have equivalent in the terrestrial plants and resembles the chemical and biological properties of mammalian glycosaminoglycans. In this perspective, are receiving growing interest for application on health-related fields. On this review, we will focus on the biomedical applications of marine algae sulfated polymers, in particular on the development of innovative systems for tissue engineering and drug delivery approaches.


Oliveira M.B.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Mano J.F.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine
Analytical Chemistry | Year: 2013

As the formation of healthy tissue and the treatment of several diseases are often dependent on an effective and prolonged action of bioactive agents, the delivery of molecules for therapeutic or induction purposes in a tissue is a common procedure. The correct administration of those agents is often dependent on tailored delivery mechanisms from hydrogel or polymeric matrixes. To the best of our knowledge, methods for the high-throughput monitoring of bioactive agent delivery are nonexistent. The methods for the in vitro monitoring of molecule release are expensive and laborious. As a simple alternative to these methods, we propose the imprinting of superhydrophobic biomimetic surfaces with ring-shaped transparent spots with concentric superhydrophobic millimetric regions to be used as bioactive agent release study platforms. We designed an array where polymeric precursors mixed with a growth-factor model protein labeled with a fluorescent tag could be dispensed in the concentric highly repellent regions and cross-linked afterward, generating a polymeric protein-loaded sphere. The ring-shaped region was then filled with a physiological-like fluid that covered the polymeric sphere. The acquisition of sequential images of each spot over time using microscopy methods allowed one to easily monitor the protein release by image-based fluorescence quantification. As the platform is easily adaptable and amenable for future automation in order to mimic standardized organ dynamics, we concluded that the device shows applicability for rapid and efficient in vitro bioactive agent release studies. © 2013 American Chemical Society.


Salgado C.L.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine
Integrative biology : quantitative biosciences from nano to macro | Year: 2012

We report on the development of a new array-based screening flat platform with the potential to be used as a high-throughput device based on biomimetic polymeric substrates for combinatorial cell/3D biomaterials screening assays in the context of tissue engineering. Polystyrene was used to produce superhydrophobic surfaces based on the so-called lotus effect. Arrays of hydrophilic regions could be patterned in such surfaces using UV/ozone radiation, generating devices onto which combinatorial hydrogel spots were deposited. The biological performance of encapsulated cells in hydrogels could be tested in an in vitro 3D environment assuming that each site was isolated from the others due to the high contrast of wettability between the patterned spots and the superhydrophobic surroundings. Three different polymers-chitosan, collagen and hyaluronic acid-were combined with alginate in different proportions in order to obtain combinatorial binary alginate-based polymeric arrays. The effect of the addition of gelatin to the binary structures was also tested. The gels were chemically analyzed by FTIR microscopic mapping. Cell culture results varied according to the hydrogel composition and encapsulated cell types (L929 fibroblast cells and MC3T3-E1 pre-osteoblast cells). Cell viability and number could be assessed by conventional methods, such as MTS reduction test and dsDNA quantification. Non-destructive image analysis was performed using cytoskeleton and nuclei staining agents and the results were consistent with the ones obtained by conventional sample-destructive techniques. Briefly, L929 cells showed higher number and viability for higher alginate-content and collagen-containing hydrogels, while MC3T3-E1 showed higher cell viability and cell number in lower alginate-content and chitosan containing hydrogels. The addition of gelatin did not influence significantly cell metabolic activity or cell number in any of the encapsulated cell types. This journal is © The Royal Society of Chemistry 2012


Pirraco R.P.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Reis R.L.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine | Marques A.P.,European Institute of Excellence on Tissue Engineering and Regenerative Medicine
Journal of Tissue Engineering and Regenerative Medicine | Year: 2013

Heterotypic cell interactions are essential for the homeostasis of bone tissue, in particular the widely studied interaction between osteoblasts and osteoclasts. Closely related with osteoclasts are monocytes/macrophages. These have been shown to produce osteogenic factors, e.g. BMP-2, which plays a key role in bone metabolism. However, the mechanisms through which monocytes/macrophages interact with osteoblasts are still elusive. The aim of this work was to assess the influence of human peripheral blood monocytes/macrophages over the early osteogenic differentiation of human bone marrow stromal cells (hBMSCs) in the presence of dexamethasone-supplemented medium. The co-cultures were performed using porous transwells that allowed the interaction between both cell types through the production of paracrine factors. The potential effect of BMP-2 produced by monocytes/macrophages was addressed by adding an anti-BMP-2 antibody to the co-cultures. hBMSCs cultured in the presence of monocytes/macrophages had a higher proliferation rate than hBMSCs monocultures. The quantification of early osteogenic marker alkaline phosphatase (ALP) revealed higher activity of this enzyme in cells in the co-culture throughout the time of culture. Both of these effects were inhibited by adding an anti-BMP-2 antibody to the cultures. Moreover, qRT-PCR for osteocalcin and osteopontin transcripts showed overexpression of both markers. Once again, the effect of monocytes/macrophages over hBMSC osteogenic differentiation was completely inhibited in the co-cultures by blocking BMP-2. The present report confirmed that monocytes/macrophages produce BMP-2, which promotes osteogenic differentiation and proliferation of hBMSCs cumulatively to dexamethasone-supplemented medium. This potentially implies that monocyte/macrophages play a stronger role in bone homeostasis than so far supposed. © 2012 John Wiley & Sons, Ltd.

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