Development of hyaluronic acid, dextrin and extracellular matrix hydrogels for cell expansion

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

Single-function approach to calibrating whatman n.º 42 filter paper based on suction versus water content relationships

Several suction–water-content (s-w) calibrations for the filter paper method (FPM) used for soil-suction measurement have been published. Most of the calibrations involve a bilinear function (i.e., two different equations) with an inflection point occurring at 60 kPa

Effect of acid or alkaline catalyst and of different capping agents on the optical properties of CdS nanoparticles incorporated within a diureasil hybrid matrix

CdS nanoparticles (NPs) were synthesized using colloidal methods and incorporated within a diureasil hybrid matrix. The surface capping of the CdS NPs by 3-mercaptopropyltrimethoxysilane (MPTMS) and 3-aminopropyltrimethoxysilane (APTMS) organic ligands during the incorporation of the NPs within the hybrid matrix has been investigated. The matrix is based on poly(ethylene oxide)/poly(propylene oxide) chains grafted to a siliceous skeleton through urea bonds and was produced by sol–gel process. Both alkaline and acidic catalysis of the sol–gel reaction were used to evaluate the effect of each organic ligand on the optical properties of the CdS NPs. The hybrid materials were characterized by absorption, steady-state and time-resolved photoluminescence spectroscopy and High Resolution Transmission Electron Microscopy (HR-TEM). The preservation of the optical properties of the CdS NPs within the diureasil hybrids was dependent on the experimental conditions used. Both organic ligands (APTMS and MPTMS) demonstrated to be crucial in avoiding the increase of size distribution and clustering of the NPs within the hybrid matrix. The use of organic ligands was also shown to influence the level of interaction between the hybrid host and the CdS NPs. The CdS NPs showed large Stokes shifts and long average lifetimes, both in colloidal solution and in the xerogels, due to the origin of the PL emission in surface states. The CdS NPs capped with MPTMS have lower PL lifetimes compared to the other xerogel samples but still larger than the CdS NPs in the original colloidal solution. An increase in PL lifetimes of the NPs after their incorporation within the hybrid matrix is related to interaction between the NPs and the hybrid host matrix.

Biological behaviour of thin films consisting of Au nanoparticles dispersed in a TiO2 dielectric matrix

In this work it was studied the possible use of thin films, composed of Au nanoparticles (NPs) embedded in a TiO2 matrix, in biological applications, by evaluating their interaction with a well-known protein, Bovine Serum Albumin (BSA), as well as with microbial cells (Candida albicans). The films were produced by one-step reactive DC magnetron sputtering followed by heat-treatment. The samples revealed a composition of 8.3 at.% of Au and a stoichiometric TiO2 matrix. The annealing promoted grain size increase of the Au NPs from 3 nm (at 300 °C) to 7 nm (at 500 °C) and a progressive crystallization of the TiO2 matrix to anatase. A broad localized surface plasmon resonance (LSPR) absorption band (λ = 580–720 nm) was clearly observed in the sample annealed at 500 °C, being less intense at 300 °C. The biological tests indicated that the BSA adhesion is dependent on surface nanostructure morphology, which in turn depends on the annealing temperature that changed the roughness and wettability of the films. The Au:TiO2 thin films also induced a significant change of the microbial cell membrane integrity, and ultimately the cell viability, which in turn affected the adhesion on its surface. The microstructural changes (structure, grain size and surface morphology) of the Au:TiO2 films promoted by heat-treatment shaped the amount of BSA adhered and affected cell viability.

Acetylated bacterial cellulose coated with urinary bladder matrix as a substrate for retinal pigment epithelium

This work evaluated the effect of acetylated bacterial cellulose (ABC) substrates coated with urinary bladder matrix (UBM) on the behavior of Retinal Pigment Epithelium (RPE), as assessed by cell adhesion, proliferation and development of cell polarity exhibiting transepithelial resistance and polygonal shaped-cells with microvilli. Acetylation of bacterial cellulose (BC) generated a moderate hydrophobic surface (around 65°) while the adsorption of UBM onto these acetylated substrates did not affect significantly the surface hydrophobicity. The ABS substrates coated with UBM enabled the development of a cell phenotype closer to that of native RPE cells. These cells were able to express proteins essential for their cytoskeletal organization and metabolic function (ZO-1 and RPE65), while showing a polygonal shaped morphology with microvilli and a monolayer configuration. The coated ABC substrates were also characterized, exhibiting low swelling effect (between 1.52.0 swelling/mm3), high mechanical strength (2048 MPa) and non-pyrogenicity (2.12 EU/L). Therefore, the ABC substrates coated with UBM exhibit interesting features as potential cell carriers in RPE transplantation that ought to be further explored.

Investigating a galactomannan gel obtained from Cassia grandis seeds as immobilizing matrix for Cramoll lectin

Characterization, with emphasis on the rheological properties, of Cassia grandis seeds galactomannan gel containing immobilized Cramoll 1,4 is presented. The gels, with and without immobilized Cramoll 1,4, were evaluated along time by rheometry, pH, color, microbial contamination and lectin hemagglutinating activity (HA). Rheological determinations confirmed the gels to be very stable up to 30 days with variations occurring after this period. Rheological data also showed that the gel/Cramoll 1,4 immobilizing matrix loses its elastic modulus substantially after 60 days. Both gels presented no microbial contamination as well as a pH close to neutral. Colorimetric parameters demonstrated the gels transparency with occasional yellowness. The opacity of the galactomannan gel did not change significantly along the study; the same did not occur for the gel with immobilized Cramoll 1,4 as a statistically significant reduction of its opacity was observed. In what concerns immobilized Cramoll 1,4HA, up to 90% of its initial HA was maintained after 20 days, with a decrease to 60% after 60 days. These results combined with the thickening and stabilizing characteristics of the galactomannan gel make this gel a promising immobilizing matrix for Cramoll 1,4 that can be further exploited for clinical and cosmetic applications.

Enalapril alters the formation of the collagen matrix in spontaneously hypertensive rats

OBJECTIVE: To assess the effect of the inhibition of the angiotensin-converting enzyme on the collagen matrix (CM) of the heart of newborn spontaneously hypertensive rats (SHR) during embryonic development. METHODS: The study comprised the 2 following groups of SHR (n=5 each): treated group - rats conceived from SHR females treated with enalapril maleate (15 mg. kg-1.day-1) during gestation; and nontreated group - offspring of nontreated females. The newborns were euthanized within the first 24 hours after birth and their hearts were removed and processed for histological study. Three fields per animal were considered for computer-assisted digital analysis and determination of the volume densities (Vv) of the nuclei and CM. The images were segmented with the aid of Image Pro Plus® 4.5.029 software (Media Cybernetics). RESULTS: No difference was observed between the treated and nontreated groups in regard to body mass, cardiac mass, and the relation between cardiac and body mass. A significant reduction in the Vv[matrix] and a concomitant increase in the Vv[nuclei] were observed in the treated group as compared with those in the nontreated group. CONCLUSION: The treatment with enalapril of hypertensive rats during pregnancy alters the collagen content and structure of the myocardium of newborns.

Importância da interação entre a integrina Mac-1 dos leucócitos e a glicoproteína Iba das plaquetas para o recrutamento de leucócitos pelas plaquetas e para a resposta inflamatória à lesão vascular

OBJETIVO: Avaliar a importância da interação entre a integrina Mac-1 dos leucócitos (a Mb 2) e a glicoproteína (GP) Iba das plaquetas para o recrutamento de leucócitos após a lesão vascular e o efeito da neutralização da interação Mac-1-GPIba sobre a proliferação celular e a hiperplasia neointimal desencadeadas por lesão vascular. MÉTODOS: Um peptídeo denominado M2 ou anticorpo anti-M2 foi desenvolvido para bloquear a interação Mac-1-GPIba . Esse peptídeo foi injetado e comparado com anticorpo-controle em camundongos C57B1/6J submetidos a lesão vascular da artéria femoral com corda-guia. Um, cinco ou 28 dias após a lesão vascular, as artérias femorais foram retiradas para a realização de morfometria e imuno-histoquímica. RESULTADOS: O bloqueio da interação Mac-1-GPIba promoveu uma redução estatisticamente significativa do número de leucócitos na camada média no primeiro dia após a lesão vascular (controle: 7,9±5,0% do total de células versus anti-M2: 2,0±1,6%, p=0,021), bem como determinou uma diminuição estatisticamente significativa do acúmulo de leucócitos na neoíntima em cinco e 28 dias (controle: 42,3±12,9% versus anti-M2: 24,6±10,8%, p=0,047 e controle: 7,9±3,0% versus anti-M2: 3,3±1,3%, p=0,012; respectivamente). A proliferação celular na camada média do vaso em cinco dias pós-lesão foi reduzida com o bloqueio da interação Mac-1-GPIba (controle: 5,0±2,9% do total de células versus anti-M2: 1,8±0,5%; p=0,043), assim como houve diminuição significativa da proliferação celular na camada íntima do vaso em 28 dias (controle: 3,8±1,7% versus anti-M2: 2,0±1,2%; p=0,047). O bloqueio da interação Mac-1-GPIba também determinou uma redução estatisticamente significativa do espessamento intimal em 28 dias pós-lesão (controle: 10.395±3.549 µm² versus anti-M2: 4.561±4.915 µm²; p=0,012). CONCLUSÃO: O recrutamento de leucócitos após a lesão vascular é dependente da interação Mac-1-GPIba e a neutralização dessa interação inibe a proliferação celular e a formação neointimal.

Influence of Spironolactone on Matrix Metalloproteinase-2 in Acute Decompensated Heart Failure

Background: Matrix metalloproteinases (MMPs) are a family of enzymes important for the resorption of extracellular matrices, control of vascular remodeling and repair. Increased activity of MMP2 has been demonstrated in heart failure, and in acutely decompensated heart failure (ADHF) a decrease in circulating MMPs has been demonstrated along with successful treatment. Objective: Our aim was to test the influence of spironolactone in MMP2 levels. Methods: Secondary analysis of a prospective, interventional study including 100 patients with ADHF. Fifty patients were non-randomly assigned to spironolactone (100 mg/day) plus standard ADHF therapy (spironolactone group) or standard ADHF therapy alone (control group). Results: Spironolactone group patients were younger and had lower creatinine and urea levels (all p < 0.05). Baseline MMP2, NT-pro BNP and weight did not differ between spironolactone and control groups. A trend towards a more pronounced decrease in MMP2 from baseline to day 3 was observed in the spironolactone group (-21 [-50 to 19] vs 1.5 [-26 to 38] ng/mL, p = 0.06). NT-pro BNP and weight also had a greater decrease in the spironolactone group. The proportion of patients with a decrease in MMP2 levels from baseline to day 3 was also likely to be greater in the spironolactone group (50% vs 66.7%), but without statistical significance. Correlations between MMP2, NT-pro BNP and weight variation were not statistically significant. Conclusion: MMP2 levels are increased in ADHF. Patients treated with spironolactone may have a greater reduction in MMP2 levels.

Untersuchung der Auswirkung der MAC-Zugriffsverfahren im IEEE 802.11 auf die L4-Performance in mobilen IP-Netzen

Gegenstand der hier vorliegenden Arbeit ist die Untersuchung der Auswirkung der MAC-Zugriffsverfahren CSMA/CA auf das Transportschicht-Protokoll in WLAN-Netzen. Zum Erreichen des Ziels dieser Arbeit wurde es zuerst nachgewiesen, dass CSMA/CA zum TCP-Retransmission führen könnte. Dann die kumulative Verteilung der OWDs auf jedem Szena-rio analysiert. Am Ende wurde die kumulative Verteilung der Abstände zwischen den Spitzenwerten der OWD nach der Datenverarbeitung auf jedem Szenario analysiert.

Freshwater sponges (Porifera, Demospongiae) in a benthic filter feeding community at the Guanacaste Dry Forest, Costa Rica

The freshwater sponges Trochospongilla variabilis Bonetto & Ezcurra de Drago (1973), Radiospongilla crateriformis (Potts, 1882), Spongilla cenota Penney & Racek (1968) and Corvoheteromeyenia heterosclera (Ezcurra de Drago, 1974) compose with the sphaerid bivalve Eupera cubensis (Prime, 1865) and several Phylactolaemata bryozoans a benthic filter feeding community living in seasonal lentic and lotic habitats with high Particulate Organic Carbon (POC), low conductivity and acid pH within the Costa Rica Dry Forest biome. The sponge specimens gathered led to the re-description of the four species.

Contribuição ao estudo dos Embiídeos: IV. Polimorfismo sexual da região cefálica de Embolyntha batesi Mac lachlan, 1877 (Embiidina, Embiidae)

Em continuação as pesquisas que vimos realizando nos Embiídeos é feito um estudo comparado das peças bucais entre machos e fêmeas de Embolyntha batesi. A cabeça é prognata recoberta por diminutas cerdas. É a região mais resistente do inseto, devido proteger, além de outros órgãos, principalmente, o sistema nervoso. Varia de tamanho nos dois sexos com os índices (comprimento : largura) na fêmea de 1,06 e nos machos de 1,36; a cabeça da fêmea é achatada, enquanto que a dos machos é alongada. Quase tôdas as suturas são visíveis nos sexos, com excessão de algumas, como é o caso da coronal e post-frontal dos machos. De tôdas as suturas, a temporal é a mais interessante, limita a região do vertex com as genas, ao mesmo tempo que origina um sulco profundo, que penetra na cápsula craniana fazendo parte do esqueleto interno da cabeça, e sendo responsável pelo aspecto diferente das mesmas. A sutura temporal, na região ventral, separa as genas das subgenas. A sutura hipostomal, em ambos os sexos, é muito acentuada, e na sua parte mais interna, vêm se inserir os ramos posteriores do tentório, e, ainda lateralmente, as maxilas. O tentório é primitivo, tendo um corpo central, de forma quadrangular e, de cada ângulo parte um ramo; dois anteriores, menores, que se dirigem para a região dorsal onde se bifurcam, indo ter próximo á base das antenas e mandíbulas, e dois ramos posteriores que seguem a direção ventral, indo ter á região hipostomal. As antenas são filiformes, variando o número de segmentos. Os olhos dos machos são reniformes, salientes e grandes, enquanto que os das fêmeas são pequenos, ovais e achatados. O número de omatídeos de macho é 34, e, na fêmea é 41, em uma determinada área. O clípeo quase não se diferencia da fronte, porém encontra-se dividido em anti-clípeo e post-clípeo. A sutura do clípeo-labro é bem acentuada, deixando transparecer, após a diafanização do material, um espessamento da cutícula na sua região mais interna, destinada a implantação dos músculos que movimentam o labro. Na parte ventral o labro apresenta sensilas, que variam quanto a forma, tamanho e estrutura nos dois sexos. As mandíbulas apresentam-se muito diferentes devida sua função, isto é, trituradora nas fêmeas e preensora nos machos. Pela simples morfologia das mandíbulas podemos identificar o sexo nos Embiídeos. Em ambos temos dentes incisivos e molares, porém mais acentuados nas fêmeas. Nos machos a região interna da mandíbula tem a forma côncava, com cutícula...

Avaluació d’un nou protocol MAC per IEEE 802.11 en un entorn multicel·lular exterior

Estudi realitzat a partir d’una estada al laboratori LSR (Logiciels, Systèmes, Réseaux) – IMAG a Grenoble, França, entre els mesos de setembre del 2005 i febrer del 2006. El grup de recerca Drakkar d’aquest laboratori va desenvolupar un nou mecanisme d’accés al medi per a xarxes d’àrea local sense fils (WLAN) anomenat Idle Sense. S’ha avaluat aquest mecanisme en entorns cel·lulars i s’ha comparat del seu comportament amb l’estàndard IEEE 802.11 DCF mitjançant la implementació en un simulador. En els entorns cel·lulars es troben condicions de transmissió molt allunyades de les ideals, i de les analitzades fins al moment, que consisteixen en escenaris formats per una única cel·la aïllada. Per aquest motiu, primer s’ha estudiat els mecanismes d’accés al medi d’Idle Sense en una cel·la i en condicions de transmissió adverses. Després, s’ha estudiat en entorns multicel·lulars, on les comunicacions entre estacions es veuen influenciades pel problema de solapament de cel·les i per les interferències provocades per les transmissions d’estacions d’altres cel·les. També s’ha comparat els resultats obtinguts amb el comportament d’IEEE 802.11 DCF i d’altres propostes de millora: Asymptotically Optimal Backoff (AOB) i Slow Decrease. Finalment, s’ha realitzat una modificació sobre Idle Sense, que s’ha anomenat Weighted Idle Sense, que resulta més adecuat per a treballar en xarxes d’àrea local sense fils en mode infraestructura.

Determinació de la ubicació estructural dels ions Yb3+ i Nb3+ en Yb:RTP, Nb:RTP i (Nb,Yb):RTP

Estudi elaborat a partir d’una estada al Stony Brook University al juliol del 2006. El RbTiOPO4 (RTP) monocristal•lí és un material d' òptica no lineal molt rellevant i utilitzat en la tecnologia làser actual, químicament molt estable i amb unes propietats físiques molt destacades, entre elles destaquen els alts coeficients electro-òptics i l'alt llindar de dany òptic que presenta. En els últims anys s’està utilitzant tecnològicament en aplicacions d'òptica no lineal en general i electro-òptiques en particular. En alguns casos ja ha substituït, millorant prestacions, a materials tals com el KTP o el LNB(1). Dopant RTP amb ions lantànids (Ln3+) (2-4), el material es converteix en un material làser auto-doblador de freqüència, combinant les seves propietats no lineals amb les de matriu làser. El RTP genera radiació de segon harmònic (SHG) a partir d’un feix fonamental amb longituds d’ona inferiors a 990 nm, que és el límit que presenta el KTP.La determinació de la ubicació estructural i l’estudi de l'entorn local del ions actius làser és de fonamental importància per a la correcta interpretació de les propietats espectroscòpiques d’aquest material. Mesures de difracció de neutrons sobre mostra de pols cristal•lí mostren que els ions Nb5+ i Ln3+ només substitueixin posicions de Ti4+ (8-9). Estudis molt recents d'EPR (electron paramagnetic resonance) semblen indicar que quan la concentració d'ió Ln3+ es baixa, aquest ió presenta la tendència a substituir l'ió alcalí present a l'estructura (10).Després dels resultats obtinguts en el present treball a partir de la tècnica EXAFS a la instal•lació sincrotò del Brookhaven National Laboratory/State University of New York (Stony Brook) es pot concloure definitivament que els ions Nb s’ubiquen en la posició Ti (1) i que els ions Yb3+ es distribueixen paritariament en les dues posicions del Ti (1 i 2). Aquests resultats aporten una valuosa informació per a la correcta interpretació dels espectres, tant d’absorció com d’emissió, del material i per la avaluació dels paràmetres del seu comportament durant l'acció làser.