Skin-derived cell-sheets as powerful tools to engineer skin analogues

Cell/cell-extracellular matrix (ECM) dynamic interactions appear to have a major role in regulating communication through soluble signaling, directing cell binding and activating substrates that participate in the highly organized wound healing process. Moreover, these interactions are also crucial for in vitro mimicking cutaneous physiology. Herein we explore cell sheet (CS) engineering to create cellular constructs formed by keratinocytes (hKC), fibroblasts (hDFB) and dermal microvascular endothelial cells (hDMEC), to target skin wound healing but also the in vitro recreation of relevant models. Taking advantage of temperature-responsive culture surfaces, which allow harvesting cultured cells as intact sheets along with the deposited native ECM, varied combinations of homotypic and heterotypic three-dimensional (3-D) CS-based constructs were developed. Constructs combining one CS of keratinocytes as an epidermis-like layer plus a vascularized dermis composed by hDFB and hDMECs were assembled as skin analogues for advancing in vitro testing. Simultaneously both hKC and hDMEC were shown to significantly contribute to the re-epithelialization of full-thickness mice skin wounds by promoting an early epithelial coverage, while hDMEC significantly lead to increased vessels density, incorporating the neovasculature. Thus, although determined by the cellular nature of the constructs, these outcomes demonstrated that CS engineering appear as an unique technology that open the possibility to create numerous combinations of 3D constructs to target defective wound healing as well as the construction of in vitro models to further mimic cutaneous functions crucial for drug screening and cosmetic testing assays.

Bioactive nanocomposite spheres with proved shape memory capability in bone defects

Bioactive glass nanoparticles (BGNPs) promote an apatite surface layer in physiologic conditions that lead to a good interfacial bonding with bone.1 A strategy to induce bioactivity in non-bioactive polymeric biomaterials is to incorporate BGNPs in the polymer matrix. This combination creates a nanocomposite material with increased osteoconductive properties. Chitosan (CHT) is a polymer obtained by deacetylation of chitin and is biodegradable, non-toxic and biocompatible. The combination of CHT and the BGNPs aims at designing biocompatible spheres promoting the formation of a calcium phosphate layer at the nanocomposite surface, thus enhancing the osteoconductivity behaviour of the biomaterial. Shape memory polymers (SMP) are stimuli-responsive materials that offer mechanical and geometrical action triggered by an external stimulus.2 They can be deformed and fixed into a temporary shape which remains stable unless exposed to a proper stimulus that triggers recovery of their original shape. This advanced functionality makes such SMPs suitable to be implanted using minimally invasive surgery procedures. Regarding that, the inclusion of therapeutic molecules becomes attractive.  We propose the synthesis of shape memory bioactive nanocomposite spheres with drug release capability.3   1.  L. L. Hench, Am. Ceram. Soc. Bull., 1993, 72, 93-98. 2.  A. Lendlein and S. Kelch, Angew Chem Int Edit, 2002, 41, 2034-2057. 3.  Ã . J. Leite, S. G. Caridade and J. F. Mano, Journal of Non-Crystalline Solids (in Press)

Acellular Gellan-gum based bilayered structures for the regeneration of osteochondral defects: a preclinical study

 In orthopaedics, the management and treatment of osteochondral (OC) defects remains an ongoing clinical challenge. Autologous osteochondral mosaicplasty has been used as a valid option for OC treatments although donor site morbidity remains a source of concern [1]. Engineering a whole structure capable of mimicking different tissues (cartilage and subchondral bone) in an integrated manner could be a possible approach to regenerate OC defects. In our group we have been proposing the use of bilayered structures to regenerate osteochondral defects [2,3]. The present study aims to investigate the pre-clinical performance of bilayered hydrogels and spongy-like hydrogels in in vivo  models (mice and rabbit, respectively), in both subcutaneous and orthotopic models. The bilayered structures were produced from Low Acyl Gellan Gum (LAGG) from Sigma-Aldrich, USA. Cartilage-like layers were obtained from a 2wt% LAGG solution. The bone-like layers were made of 2wt% LAGG with incorporation of hydroxyapatite at 20% and 30% (w/v). Hydrogels and spongy-like were subcutaneouly implanted in mice to evaluate the inflammatory response. Then, OC defects were induced in rabbit knee to create a critical size defect (4 mm diameter and 5 mm depth), and then hydrogels and sponges implanted. Both structures followed different processing methods. The hydrogels were injected allowing in situ  crosslinking. Unlike, the spongy-like were pre-formed by freeze-drying. The studies concerning subcutaneous implantation and critical size OC defect were performed for 2 and 4 weeks time, respectively. Cellular behavior and inflammatory responses were assessed by means of histology staining and biochemical function and matrix deposition by immunohistochemistry. Additionally, both OC structures stability and new cartilage and bone formation were evaluated by using vivo- computed tomography (Scanco 80). The results showed no acute inflammatory response for both approaches. New tissue formation and integration in the adjacent tissues were also observed, which present different characteristic behaviors when comparing hydrogels and sponges response. As future insights, a novel strategy for regeneration of OC defects can be designed encompassing both, hydrogels and spongy-like structures and cellular approaches. References: 1. Espregueira-Mendes J. et al. Osteochondral transplantation using autografts from the upper tibio-fibular joint for the treatment of knee cartilage lesions. Knee Surgery, Sports Traumatology, Arthroscopy 20,1136, 2012. 2. Oliveira JM. et al, Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications: Scaffold design and its performance when seeded with goat bone marrow stromal cells. Biomaterials 27, 6123, 2006. 3. Pereira D R. et al. Gellan Gum-Based Hydrogel Bilayered Scaffolds for Osteochondral Tissue Engineering. Key Engineering Materials 587, 255, 2013.

Effect of variables on the thickness of an edible coating applied on frozen fish establishment of the concept of safe dipping time

Glazing is a technique used to retard fish deterioration during storage. This work focuses on the study of distinct variables (fish temperature, coating temperature, dipping time) that affect the thickness of edible coatings (water glazing and 1.5% chitosan) applied on frozen fish. Samples of frozen Atlantic salmon (Salmo salar) at -15, -20, and -25 °C were either glazed with water at 0.5, 1.5 or 2.5 °C or coated with 1.5% chitosan solution at 2.5, 5 or 8 °C, by dipping during 10 to 60 s. For both water and chitosan coatings, lowering the salmon and coating solution temperatures resulted in an increase of coating thickness. At the same conditions, higher thickness values were obtained when using chitosan (max. thickness of 1.41±0.05 mm) compared to water (max. thickness of 0.84±0.03 mm). Freezing temperature and crystallization heat were found to be lower for 1.5% chitosan solution than for water, thus favoring phase change. Salmon temperature profiles allowed determining, for different dipping conditions, whether the salmon temperature was within food safety standards to prevent the growth of pathogenic microorganisms. The concept of safe dipping time is proposed to define how long a frozen product can be dipped into a solution without the temperature raising to a point where it can constitute a hazard.

High strain rate constitutive modeling forhistorical structures subjected to blast loading

Doctoral Thesis Civil Engineering

Structure-property relationships in polymer nanocomposites

Tese de Doutoramento em Ciência e Engenharia de Polímeros e Compósitos

Electronic transport across linear defects in graphene

We investigate the low-energy electronic transport across grain boundaries in graphene ribbons and infinite flakes. Using the recursive Green’s function method, we calculate the electronic transmission across different types of grain boundaries in graphene ribbons. We show results for the charge density distribution and the current flow along the ribbon. We study linear defects at various angles with the ribbon direction, as well as overlaps of two monolayer ribbon domains forming a bilayer region. For a class of extended defect lines with periodicity 3, an analytic approach is developed to study transport in infinite flakes. This class of extended grain boundaries is particularly interesting, since the K and K0 Dirac points are superposed.

Comparing different methods to measure biofilm thickness: the techniques are: optical coherence tomography, confocal laser scanning microscopy and low load compression test

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica)

Numerical and experimental analysis of full scale arches reinforced with GFRP materials

In this contribution, original limit analysis numerical results are presented dealing with some reinforced masonry arches tested at the University of Minho-UMinho, PT. Twelve in-scale circular masonry arches were considered, reinforced in various ways at the intrados or at the extrados. GFRP reinforcements were applied either on undamaged or on previously damaged elements, in order to assess the role of external reinforcements even in repairing interventions. The experimental results were critically discussed at the light of limit analysis predictions, based on a 3D FE heterogeneous upper bound approach. Satisfactory agreement was found between experimental evidences and the numerical results, in terms of failure mechanisms and peak load.

Importance of contact lens power and thickness in oxygen transmissibility

PURPOSE: The aim of this work was to study the central and peripheral thickness of several contact lenses (CL) with different powers and analyze how thickness variation affects CL oxygen transmissibility. METHODS: Four daily disposable and five monthly or biweekly CL were studied. The powers of each CL were: the maximum negative power of each brand; -6.00 D; -3.00 D; zero power (-0.25 D or -0.50 D), +3.00 D and +6.00 D. Central and peripheral thicknesses were measured with an electronic thickness gauge. Each lens was measured five times (central and 3mm paracentral) and the mean value was considered. Using the values of oxygen permeability given by the manufacturers and the measured thicknesses, the variation of oxygen transmissibility with lens power was determined. RESULTS: For monthly or biweekly lenses, central thickness changed between 0.061 ± 0.002 mm and 0.243 ± 0.002 mm, and peripheral thickness varied between 0.084 ± 0.002 mm and 0.231 ± 0.015 mm. Daily disposable lenses showed central values ranging between 0.056 ± 0.0016 mm and 0.205 ± 0.002 mm and peripheral values between 0.108 ± 0.05 and 0.232 ± 0.011 mm. Oxygen transmissibility (in units) of monthly or biweekly CL ranged between 39.4 ± 0.3 and 246.0 ± 14.4 and for daily disposable lenses the values range between 9.5 ± 0.5 and 178.1 ± 5.1. CONCLUSIONS: The central and peripheral thicknesses change significantly when considering the CL power and this has a significant impact on the oxygen transmissibility. Eyecare practitioners must have this fact in account when high power plus or minus lenses are fitted or when continuous wear is considered.

Reflow soldering in an oxidant atmosphere

Dissertação de mestrado integrado em Materials Engineering

Comportamento tribológico de biocompósitos de matriz de titânio

Dissertação de mestrado integrado em Engenharia de Materiais

Optimization of magnetoelectric composites based on electroactive polymers for energy harvesting and sensor application

Tese de Doutoramento em Engenharia de Materiais.

Elaboration et caractérisation des nanocristaux de Zn F O enfouis 1-x x dans des diélectriques pour des applications en spin-électronique

Tese de Doutoramento em Ciências - Especialidade em Física

Diagnóstico e ensaio de carga de asnas tradicionais de madeira

Dissertação de mestrado Integrado em Engenharia Civil