Mesenchymal stem cells culture on poly(vinylidene fluoride) piezoelectric microspheres

Dissertação de mestrado em Biofísica e Bionanossistemas

Biotechnological approaches for yield improvement of anticancer alkaloids in the plant Catharanthus roseus

Dissertação de mestrado em Biologia Molecular, Biotecnologia e Bioempreendedorismo em Plantas

Floristic and structure of an Amazonian primary forest and a chronosequence of secondary succession

ABSTRACT The analysis of changes in species composition and vegetation structure in chronosequences improves knowledge on the regeneration patterns following land abandonment in the Amazon. Here, the objective was to perform floristic-structural analysis in mature forests (with/without timber exploitation) and secondary successions (initial, intermediate and advanced vegetation regrowth) in the Tapajós region. The regrowth age and plot locations were determined using Landsat-5/Thematic Mapper images (1984-2012). For floristic analysis, we determined the sample sufficiency and the Shannon-Weaver (H'), Pielou evenness (J), Value of Importance (VI) and Fisher's alpha (α) indices. We applied the Non-metric Multidimensional Scaling (NMDS) for similarity ordination. For structural analysis, the diameter at the breast height (DBH), total tree height (Ht), basal area (BA) and the aboveground biomass (AGB) were obtained. We inspected the differences in floristic-structural attributes using Tukey and Kolmogorov-Smirnov tests. The results showed an increase in the H', J and α indices from initial regrowth to mature forests of the order of 47%, 33% and 91%, respectively. The advanced regrowth had more species in common with the intermediate stage than with the mature forest. Statistically significant differences between initial and intermediate stages (p<0.05) were observed for DBH, BA and Ht. The recovery of carbon stocks showed an AGB variation from 14.97 t ha-1 (initial regrowth) to 321.47 t ha-1 (mature forests). In addition to AGB, Ht was also important to discriminate the typologies.

Highly functional and biodegradable nanofibrous scaffolds combined with stem cells for bone and cartilage tissue engineering

Among the various possible embodiements of Advanced Therapies and in particular of Tissue Engineering the use of temporary scaffolds to regenerate tissue defects is one of the key issues. The scaffolds should be specifically designed to create environments that promote tissue development and not merely to support the maintenance of communities of cells. To achieve that goal, highly functional scaffolds may combine specific morphologies and surface chemistry with the local release of bioactive agents. Many biomaterials have been proposed to produce scaffolds aiming the regeneration of a wealth of human tissues. We have a particular interest in developing systems based in nanofibrous biodegradable polymers1,2. Those demanding applications require a combination of mechanical properties, processability, cell-friendly surfaces and tunable biodegradability that need to be tailored for the specific application envisioned. Those biomaterials are usually processed by different routes into devices with wide range of morphologies such as biodegradable fibers and meshes, films or particles and adaptable to different biomedical applications. In our approach, we combine the temporary scaffolds populated with therapeutically relevant communities of cells to generate a hybrid implant. For that we have explored different sources of adult and also embryonic stem cells. We are exploring the use of adult MSCs3, namely obtained from the bone marrow for the development autologous-based therapies. We also develop strategies based in extra-embryonic tissues, such as amniotic fluid (AF) and the perivascular region of the umbilical cord4 (Whartonâ s Jelly, WJ). Those tissues offer many advantages over both embryonic and other adult stem cell sourcess. These tissues are frequently discarded at parturition and its extracorporeal nature facilitates tissue donation by the patients. The comparatively large volume of tissue and ease of physical manipulation facilitates the isolation of larger numbers of stem cells. The fetal stem cells appear to have more pronounced immunomodulatory properties than adult MSCs. This allogeneic escape mechanism may be of therapeutic value, because the transplantation of readily available allogeneic human MSCs would be preferable as opposed to the required expansion stage (involving both time and logistic effort) of autologous cells. Topics to be covered: This talk will review our latest developments of nanostructured-based biomaterials and scaffolds in combination with stem cells for bone and cartilage tissue engineering.

Adipogenic cell sheets to recreate in vitro adipose tissue microenvironments

Cell sheet (CS) engineering, taking advantage of cellular self-matrix organized as in native tissue, has been largely explored, including by us, for different purposes [1â 3]. Herein we propose for the ï¬ rst time, the use of human adipose stem cells (hASCs)-derived CS to create adipose tissue analogues with different levels of maturation. hASCs were cultured on UpCellTM thermo-responsive dishes for 1, 3 and 5 days under basal conditions previously established by us [3]. The inï¬ uence of pre-differentiation time and respective cell number, over CS stability and differentiation was assessed. Mechanically robust CS were only obtained with 5 days pre-differentiation period. Adipogenesis was followed along the culture assessing the variation of expression of mesenchymal (CD73, CD105 but not CD90) and adipogenic (PPARg, FABP4 and LPL) markers by ï¬ ow cytometry, immunocytochemistry and RT-PCR. Increased ratio of differentiated cells was achieved for longer pre-differentiation periods, while maturation degree was modulated by the maintenance medium. Independently of the overall CS differentiation/maturation level, 3D constructs were fabricated by stacking and further culturing 3 CS. Thus, by varying the culture conditions, different 3D adipose tissue-like microenvironments were recreated, enabling future development of new tissue engineering strategies, as well as further study of adipose tissue role in the regeneration of different tissues.

Hind limb ischemia in type 1 diabetic mice as useful tool to evaluate the neovascularization of tissue engineering constructs

Hind-limb ischemia has been used in type 1 diabetic mice to evaluate treatments for peripheral arterial disease or mechanisms of vascular impairment in diabetes [1]. Vascular deficiency is not only a pathophysiological condition, but also an obvious circumstance in tissue regeneration and in tissue engineering and regenerative medicine (TERM) strategies. We performed a pilot experiment of hind-limb ischemia in streptozotocin(STZ)-induced type 1 diabetic mice to hypothesise whether diabetes influences neovascularization induced by biomaterials. The dependent variables included blood flow and markers of arteriogenesis and angiogenesis. Type 1 diabetes was induced in 8-week-old C57BL/6 mice by an i.p. injection of STZ (50 mg/kg daily for 5 days). Hind-limb ischemia was created under deep anaesthesia and the left femoral artery and vein were isolated, ligated, and excised. The contralateral hind limb served as an internal control within each mouse. Non-diabetic ischaemic mice were used as experiment controls. At the hind-limb ischemia surgical procedure, different types of biomaterials were placed in the blood vessels gap. Blood flow was estimated by Laser Doppler perfusion imager, right after surgery and then weekly. After 28 days of implantation, surrounding muscle was excised and evaluated by histological analysis for arteriogenesis and angiogenesis. The results showed that implanted biomaterials were promote faster restoration of blood flow in the ischemic limbs and improved neovascularization in the diabetic mice. Therefore, we herein demonstrate that the combined model of hind-limb ischemia in type 1 diabetes mice is suitable to evaluate the neovascularization potential of biomaterials and eventually tissue engineering constructs.  

Development of advanced cell/tissue culture systems, based on enhanced polymeric scaffolds and sophisticated bioreactors, for tissue engineering applications

Programa Doutoral em Engenharia Biomédica

Possibilidades de reabilitação urbana em bairros sociais: estudo de caso: Bairro Social de Santa Tecla, Braga

Dissertação de mestrado integrado em Engenharia Civil

Semipermeable capsules wrapping a multifunctional and self-regulated co-culture microenvironment for osteogenic differentiation

A new concept of semipermeable reservoirs containing co-cultures of cells and supporting microparticles is presented, inspired by the multi-phenotypic cellular environment of bone. Based on the deconstruction of the â stem cell nicheâ , the developed capsules are designed to drive a self-regulated osteogenesis. PLLA microparticles functionalized with collagen I, and a co-culture of adipose stem (ASCs) and endothelial (ECs) cells are immobilized in spherical liquified capsules. The capsules are coated with multilayers of poly(L-lysine), alginate, and chitosan nano-assembled through layer-by-layer. Capsules encapsulating ASCs alone or in a co-culture with ECs are cultured in endothelial medium with or without osteogenic differentiation factors. Results show that osteogenesis is enhanced by the co-encapsulation, which occurs even in the absence of differentiation factors. These findings are supported by an increased ALP activity and matrix mineralization, osteopontin detection, and the up regulation of BMP-2, RUNX2 and BSP. The liquified co-capsules also act as a VEGF and BMP-2 cytokines release system. The proposed liquified capsules might be a valuable injectable self-regulated system for bone regeneration employing highly translational cell sources.

Valorização patrimonial e urbana de Braga através da criação de circuitos turísticos

Projeto de mestrado em Património e Turismo Cultural

Plano de revitalização do centro histórico de Braga

Dissertação de mestrado em Políticas Comunitárias e Cooperação Territorial

Biomedical applications of human cathelicidin

[Excerpt] Antimicrobial peptides (AMPs) are good candidates to treat burn wounds, a major cause of morbidity, impaired life quality and resources consumption in developed countries. Tuberculosis (TB), a disease caused by the human pathogen Mycobacterium tuberculosis, represents the second world’s deadliest infectious disease, affecting around 9 million people worldwide in 2013. Of those, about 1.1 million died from the disease. The potential of cathelicin, a human AMP, in the treatment of mycobacteriosis and wound regeneration was assessed in pre-clinical trials. (...)

O eletrocardiograma de alta resolução da ativação atrial em pacientes com ou sem fibrilação atrial paroxística

OBJETIVO: Analisar os parâmetros do eletrocardiograma de alta resolução da onda P no domínio do tempo (ECGAR-P) e compará-los com: a duração da onda P no eletrocardiograma clássico (P no ECG), o diâmetro atrial esquerdo (AE) e a fração de ejeção do ventrículo esquerdo (FE) obtidos no ecocardiograma, para avaliar pacientes com fibrilação atrial paroxística (FAP). MÉTODOS: Foram estudados 181 pacientes: 117 com FAP comprovada e 64 sem FAP. Os parâmetros do ECGAR-P foram: a duração da onda P filtrada (DPF), as voltagens da raiz quadrada média dos últimos 40, 30 e 20 ms da onda P filtrada (RMS 40, RMS 30 e RMS 20), a voltagem da raiz quadrada média dos potenciais da onda P filtrada (RMS P), a integral dos potenciais da onda P filtrada (Integral P) e a duração dos potenciais tardios da onda P filtrada abaixo de 3 µV (PT<3). RESULTADOS: Os parâmetros que apresentaram diferenças estatisticamente significantes entre os grupos foram: DPF, RMS 40, 30 e 20, PT<3, P no ECG e AE. Os cálculos pela curva ROC mostraram, para cada parâmetro, o melhor valor de corte e os estimadores de desempenho: sensibilidade, especificidade, área sob a curva e p-value (p) ou nível descritivo. CONCLUSÃO: O ECGAR-P no domínio do tempo mostrou-se melhor que o eletrocardiograma clássico e o ecocardiograma para identificar pacientes com fibrilação atrial paroxística.

An Investigation into Irish State patronage of the visual arts.

The focus of this research is in ascertaining how and why the Irish state patronises the visual arts. The framework that the state puts in place for the support of the arts influences the creation of art. The initial standpoint from which to evaluate the system is founded on the belief that art is of social good. The second source of belief is in the necessity for an autonomous setting in which it can be created. These two beliefs underpin state patronage of the arts in contemporary society. Therefore they have to be examined carefully in order to see if they hold up. This requires an investigation into the formation of value. This is undertaken by looking into the social development of western society and its influence on the placement of arts value. Establishing how arts value to society is defined provides a means by which to investigate the manner in which the state patronises the visual arts. Two case studies provide the evidence in how the state supports the arts and why it chooses to do so. The Irish Museum of Modem Art is used as an example of the state’s role in the maintenance of the canon of art. The second case study looks at the work being done by Breaking Ground. Breaking Ground is an extensive art project as an element of the regeneration process happening in Ballymun funded by the state. It provides and insight into how the state utilises art in the unification of a social group.

Ireland’s understudied flatfish: reproduction, age and growth of the dab Limanda limanda (L.) in Irish coastal waters

In Ireland, although flatfish form a valuable fishery, little is known about the smallest, the dab Limanda limanda. In this study, a variety of parameters of reproductive development, including ovarian phase description, gonadosomatic index (GSI), hepatosomatic index (HSI), relative condition (Kn) and oocyte size were analysed to provide information on the dab’s reproductive cycle and spawning periods. Sampling were collected monthly over an 18-month period using bottom trawls of the Irish coastline. A six phase macroscopic guide was developed for both sexes of dab, and verified using histology. In comparisons of macroscopic and microscopic phases, there was high agreement in the proposed female guide (86%), with males demonstratively lower (62%). No significant bias was observed between the the two reproductive methods. When the male macroscopic guide was examined, misclassification was high in phase 5 and phase 5 (41%), with 96% of misclassification occurring in adjacent phases. The sampled population was primarily composed of females, with ratios of females to males 1:0.6, although the predominance of females was less noticeable during the reproductive season. Oocyte growth in dab follows asynchronous development, and spawn over a protracted period indicating a batch spawning strategy. Spawning occurred mainly in early spring, with total regeneration of gonads by May. The length at which 50% of the population was reproductively mature was identified as 14cm and 17cm, for male and female dab, respectively. Precision and bias in age determinations using whole otoliths to age dab was investigated using six age readers from various institutions. Low levels of precision were obtained (CV: 10-23%) inferring the need for an alternative methodology. Precision and bias was influence by the level of experience of the reader, with ageing error attributed to interpretative differences and difficulty in edge determination. Sectioned otolith age determinations were subsequently compared to whole otolith age determinations using two age readers experienced in dab ageing. Although increased precision was observed in whole otoliths from previous estimates (CV=0%, 0% APE), sectioned otoliths were used for growth models. This was based on multinominal logistic regression on age length keys developed using both ageing methods. Biological data (length and age) for both sexes was applied to four growth models, where the Akaike criterion and Multi model Inference indicated the logistic model as having the best fit to the collected data. In general, female dab attained a longer length then males, with growth rates significantly different between the two sexes. Length weight relationships between the two sexes were also significantly different.