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Made available in DSpace on 2014-12-17T15:42:27Z (GMT). No. of bitstreams: 1 MariaSPL.pdf: 2188057 bytes, checksum: 3142bea2af0c353bf960b7b878efd707 (MD5) Previous issue date: 2006-11-17 / The aim of this study was to generate an asymmetric biocompactible and biodegradable chitosan membrane modified by the contact with a poly(acrylic acid) solution at one of its sides at room temperature and 60◦C. The pure chitosan membrane, as well as the ones treated with poly(acrylic acid) were characterized by infrared spectroscopy (FTIRATR) at angles of 39◦, 45◦ and 60◦ , swelling capacity in water, thermal analysis (TG/DTG), scanning electronic microscopy (SEM) and permeation experiments using metronidazole at 0,1% and 0,2% as a model drug. The results confirmed the presence of ionic interaction between chitosan and poly(acrylic acid) by means of a polyelectrolyte complex (PEC) formation. They also showed that such interactions were more effective at 60◦C since this temperature is above the chitosan glass transition temperature wich makes the diffusion of poly(acrylic acid) easier, and that the two treated membranes were asymmetrics, more thermically stable and less permeable in relation to metronidazole than the pure chitosan membrane / O prop?sito deste estudo foi produzir uma membrana assim?trica biocompat?vel e biodegrad?vel de quitosana modificada pelo contato com uma solu??o de poli(?cido acr?lico) em uma de suas superf?cies ? temperatura ambiente e a 60◦C. As membranas de quitosana pura, quitosana com poli(?cido acr?lico) a 25◦C e quitosana com poli(?cido acr?lico) a 60◦C foram caracterizadas por: espectroscopia no infravermelho (FTIRATR) em ?ngulos de incid?ncia de 39◦, 45◦ e 60◦, ganho de massa em ?gua, an?lise t?rmica (TG/ DTG), microscopia eletr?nica de varredura (MEV) e, ainda, atrav?s dos ensaios de permea??o in vitro utilizando como f?rmaco modelo o metronidazol em solu??o aquosa nas concentra??es de 0,1 e 0,2%. Os resultados obtidos comprovaram a exist?ncia de intera??es i?nicas entre os dois pol?meros, atrav?s da forma??o dos chamados complexos polieletrol?ticos. Tamb?m mostraram que a reticula??o foi mais efetiva a 60◦C , uma vez que essa temperatura est? acima da temperatura de transi??o v?trea da quitosana, o que facilita a difus?o do poli(?cido acr?lico) e que as membranas resultantes s?o assim?tricas, mais est?veis termicamente e menos perme?veis ao metronidazol do que a membrana de quitosana pura

Komplexe aus entgegengesetzt geladenen Polyelektrolyten haben sowohl in der Biologie als auch in den Materialwissenschaften eine große Bedeutung. Im Mittelpunkt des Interesses stehen besonders die Kondensation der DNA in vitro, die Struktur des Nukleosoms im Zellkern, nicht-virale Systeme zur Transfektion von DNA in Zellen oder der Vorgang der layer-by-layer Adsorption. Verschiedene Theorien befassen sich mit den treibenden Kräften solcher Komplexbildungen. Allerdings standen experimentelle Untersuchungen auf diesem Gebiet bisher noch aus. Dieser Arbeit liegt die Fragestellung zu Grunde, ob es mit Hilfe der Rasterkraftmikroskopie möglich ist, die Struktur einzelner Polyelektrolytkomplexe, bestehend aus den beiden Polyelektrolyten DNA und dendronisierten Polymer, aufzuklären und ihre Komplexbildung zu untersuchen. Die Komplexe bildeten sich in Lösung und wurden anschließend auf einer unbeschichteten oder mit positiven Polymeren beschichteten Glimmeroberfläche adsorbiert. Auf der positiv beschichteten Glimmeroberfläche hafteten DNA-dendronisierte Polymer Komplexe mit einem Ladungsverhältnis von 1:1 bis 1:0.7 (DNA:dendronisiertes Polymer). Anhand der hochaufgelösten rasterkraftmikroskopischen Aufnahmen wurde ein Modell entwickelt, das die Umwicklung der DNA um das dendronisierte Polymer beschreibt. Der DNA-DNA Abstand ergab sich zu (2.30 ± 0.27) nm für den Komplex mit DNA und zweiter Generation dendronisierter Polymere und zu (2.16 ± 0.27) nm mit vierter Generation. Die theoretische Vorhersage der Überladung der Komplexe konnte experimentell bestätigt werden. Mit Hilfe der Rasterkraftmikroskopie konnte überdies der Einfluss des Salzgehaltes der Lösung auf die Bildung der Komplexe mit DNA und zweiter Generation dendronisierter Polymere untersucht werden. Wie man anhand des Zusammenwirkens von elektrostatischen Kräften und entropischen Wechselwirkungen bei der Adsorption von Polyelektrolyten vorhersagen kann, durchlief der DNA-DNA Abstand ein Minimum bei ansteigendem Salzgehalt. Bei sehr hohem Salzgehalt (2.4 M NaCl) konnte das Ablösen der DNA von dem Komplex beobachtet werden. Die untersuchten DNA/dendroniserten Polymer Komplexe bilden ein neues Modellsystem, mit dem einzelne Polyelektrolyt-Wechselwirkungen direkt untersucht werden können. Ein Vergleich der experimentellen Daten mit den vorhandenen Theorien zeigte, dass der Prozess des Überladens weitgehend durch elektrostatische Wechselwirkung zwischen den beiden Polyelektrolyten beschrieben werden kann. Sowohl entropische Beiträge als auch die Biegeenergie der umwickelnden DNA sind vernachlässigbar. Basierend auf diesen Ergebnissen können neue Trägerstrukturen für effizientere nicht-virale DNA-Transfektionssysteme entwickelt werden. / Complexes of oppositely charged polyelectrolytes play an important role in both biology and material science, for instance DNA condensation in vitro, nucleosomal structure, non-viral gene transfection systems as well as layer-by-layer adsorption. Although there are theories predicting overcharging of polyelectrolyte complexes, the driving forces are still under debate and systematic experimental studies on single polyelectrolytes remain challenging. Therefore the question arose if it is possible to analyze single polyelectrolyte complexes, using DNA and dendronized polymers, with the scanning force microscope in order to investigate the complexation in detail. For the complex analysis, the polyelectrolytes were allowed to interact in solution and then to adsorb on negatively charged mica or on mica coated with a positively charged polymer. Scanning force microscopy was used to investigate the adsorbed species. DNA/dendronized polymer complexes of charge ratio of 1/1 through 1/0.7 adsorbed on mica coated with a positively charged polymer. The analysis of high resolution molecular images indicated that DNA wraps around the dendronized polymer with an estimated pitch of (2.30 ± 0.27) nm and (2.16 ± 0.27) nm for dendronized polymers of generation two and four, respectively. In the proposed model the polyelectrolyte with the smaller linear charge density is wrapped around the more highly charged dendronized polymer, resulting in a negatively overcharged complex. This overcharging is consistent within recent theories of spontaneous overcharging of complexes of one polyelectrolyte wrapping around the other. Using the complex of DNA and dendronized polymers of second generation, the influence of monovalent salt concentration on the molecular structure was studied. By increasing the salt concentration the pitch showed a minimum as predicted by the interplay of electrostatic forces and entropic interactions of polyelectrolyte adsorption. At high salt concentration (2.4 M NaCl) the release of DNA from the complex can be observed. The results showed that the DNA/dendronized polymer system can be used as a new, high potential model system to investigate single polyelectrolyte interactions. With regard to recent theories, the experimental results indicate that the overcharging of the complex is mainly driven by electrostatic forces whereas contributions of counterion entropy and bending energy seem to be negligible. This understanding may be useful for the design of single polyelectrolyte complexes for non-viral gene delivery systems and might help to optimize the transfection efficiency based on the structure of the vector system.

<p>Synthetic carbohydrate carrying polymers have gained substantial attention recently due to their biocompatibility and their wide range of applications such as targeted drug delivery systems, gene therapy and cell-specific biomedical materials. The overall objective of this research is to develop novel carbohydrate bearing polymers through modification of polyvinylamine (PVAm) backbone with galactose groups and to discover potential applications for this new category of glycopolymers.</p> <p>PVAm-g-galactose (PVAm-GAL) with various molecular weights and grafting extents were prepared and characterized by nuclear magnetic resonance (NMR) and potentiometric and conductometric titration. Bonding of PVAm-GAL with phenylboronic acid modified PVAm (PVAm-PBA) and phenylboronate modified surfaces were studied on a quartz crystal microbalance with dissipation (QCM-D). Multilayer assembly of alternating layers of PVAm-GAL and PVA-PBA was formed on the silica sensor.</p> <p>Interaction of PVAm-GAL with RCA₁₂₀ lectin, a galactose specific protein, was studied on a silica sensor using QCM-D. Galactose binding proteins are overexpressed in hepatocyte and have been widely exploited for targeting the liver tissue with the help of galactosylated polymeric carriers. RCA₁₂₀lectin shows spontaneous adsorption on galactose rich surfaces obtained by the adsorption of PVAm-GAL on silica sensors. Association constant of the interaction was calculated . Effect of pretreatment with bovine serum albumin (BSA) was also examined.</p> <p>Cationic polymers can form polyelectrolyte complexes (PECs) with negatively charged DNA, resulting in formation of nano-sized complexes for gene delivery purposes. PECs based on PVAm-GAL and different DNA samples were prepared and their physicochemical properties were investigated using dynamic light scattering (DLS) and electrophoretic mobility measurements. Furthermore, PVAm-GAL was studied as coating for Ca-alginate beads which are widely used for cell encapsulation purposes. PVAm-GAL can strengthen the capsule’s surface and increase the physicochemical stability of the beads against chemical degradations. PVAm-GAL coated alginate beads successfully survived treatment with sodium citrate and high ionic strength solutions.</p> / Master of Applied Science (MASc)

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / Este trabalho tem como objetivo a sÃntese e caracterizaÃÃo de nano e micropartÃculas para liberaÃÃo de fÃrmaco para tratamento da malÃria. Microesferas de quitosana de alta massa molar (QTa) e goma do chichà (CH) foram sintetizadas por complexaÃÃo polieletrolÃtica e reticuladas com glutaraldeÃdo. Os diÃmetros das microesferas reticuladas e nÃo-reticuladas foram de 544 &#61617; 3 &#956;m e 558 &#61617; 2 &#956;m, respectivamente. As esferas reticuladas nÃo foram solÃveis em meio Ãcido (pH 1,2). Ensaios de intumescimento mostraram que as microesferas intumesciam mais em pH 1,2 do que em pH 7,4 e que as reticuladas possuÃam menor intumescimento do que as nÃo reticuladas. A liberaÃÃo sequenciada de cloroquina, a partir das microesferas, foi realizada por 2 h em pH 1,2 seguida por uma liberaÃÃo em pH 7,4. A microesfera reticulada liberou 64% da cloroquina em pH 1,2, com um total do fÃrmaco liberado de 92%. O perfil de liberaÃÃo da mesma amostra em pH 7,4 apresenta uma liberaÃÃo controlada do fÃrmaco por cerca de 50 h. NanopartÃculas de QT e CH foram produzidas utilizando como rotas de sÃntese a complexaÃÃo polieletrolÃtica e formaÃÃo de base de Schiff. Na formaÃÃo de complexos polieletrolÃticos parÃmetros como massa molar da quitosana, razÃo molar de carga (n+/n-), ordem de adiÃÃo e concentraÃÃo dos polieletrÃlitos influenciam no tamanho, potencial zeta, Ãndice de polidispersividade e estabilidade das nanopartÃculas em soluÃÃo. O potencial zeta das partÃculas com excesso de QT à positivo e quando se diminui a razÃo molar de carga (n+/n-) para 0,1 o potencial torna-se negativo devido o excesso de chichÃ. Os diÃmetros das nanopartÃculas variaram de 80 a 1200 nm dependendo da concentraÃÃo dos polieletrÃlitos e da quitosana utilizada. NanopartÃculas formadas por quitosana de baixa massa molar (QTb) possuem tamanho maior do que as formadas por quitosana de alta massa molar (QTa). Quando a razÃo de cargas (n+/n-) e a concentraÃÃo dos polieletrÃlitos diminuem o tamanho das nanopartÃculas tambÃm diminui. Na liberaÃÃo da cloroquina em matrizes de CH, QTa e QTb de razÃo 5 e 0,1 duraram cerca 15 dias liberando atà 99% do fÃrmaco, porÃm apenas a razÃo de cargas influenciou no perfil da liberaÃÃo. NanopartÃculas formadas via base de Schiff foram preparadas. A influÃncia de parÃmetros tais como: grau de oxidaÃÃo da goma do chichÃ, massa molar da quitosana, ordem de adiÃÃo e razÃo entre as massas dos polissacarÃdeos foram investigados em relaÃÃo ao tamanho, potencial zeta e estabilidade. O potencial zeta mostrou-se positivo para partÃculas com excesso de QT e negativo para partÃculas com excesso de CH. Os diÃmetros das partÃculas variaram de 30 a 450 nm, dependendo do grau de oxidaÃÃo do CH e da massa molar de QT. Para goma do chichà com menor grau de oxidaÃÃo nanopartÃculas de QTa apresentaram-se maiores do que as formadas por QTb, e para a goma com maior grau de oxidaÃÃo nanopartÃculas de QTb possuÃam maiores tamanhos do que as formadas por QTa. / The aim of this work was the synthesis and characterization of nano and microparticles for malaria drug delivery system. Chitosan microspheres of high molar mass (QTa) and chichà gum (CH) were synthesized by polyelectrolyte complexation and crosslinked with glutaraldehyde. The diameters of the microspheres crosslinked and non-crosslinked were 544 &#61617; 3 &#956;m and 558 &#61617; 2 &#956;m, respectively. The crosslinked beads were not soluble in acidic medium (pH 1.2). The swelling of microspheres was higher in pH 1.2 and that the crosslinked beads have less swelling than non-crosslinked. The sequential release of chloroquine from the microspheres was performed for 2 h followed by a release in pH 7.4. The crosslinked microsphere released 64% of chloroquine at pH 1.2, with a total of drug released of 92%. The release profile of the same sample at pH 7.4 provides a controlled release of the drug for about 50h. QT and CH nanoparticles were prepared using polyelectrolyte complexation and formation of Schiff base. In the formation of polyelectrolyte complex, parameters such as molecular weight of chitosan, the molar ratio of charge (n+/ n-), order of addition and concentration of the polyelectrolyte influence the size, zeta potential, polydispersity index and stability of the nanoparticles in solution. The zeta potential of particles in excess of QT was positive and when the charg molar ratio (n+/ n ) decreases to 0.1 the potential becomes negative due to the excess of CH. The nanoparticles diameters vary from 80 to 1,200 nm depending on the concentration of the polyelectrolyte and the chitosan used. Chitosan nanoparticles formed by a low molecular weight (QTb) were larger than those formed by chitosan of high molecular weight (QTa). The decrease of the charge ratio (n+/n-) and the polyelectrolyte concentrations lead to small size nanoparticle. The release of chloroquine in matrices of CH, QTa and QTb ratio ratio 5 and 0.1 lasted 15 days by releasing up to 99% of the drug, however only the ratio influenced the release profile. Nanoparticles formed by Schiff base reaction were produced. The influence of parameters such as degree of oxidation of CH, chitosan molar mass, addition and masses ration of polysaccharides on size, zeta potential and stability were investigated. The zeta-potential was positive for particles with an excess of QT and negative with excess CH. The particle diameters ranged from 30 to 450 nm, depending on the degree of oxidation of CH and the molar mass of QT. Particle formed with low oxidation of CH and high molar mass chitosan are bigger than those formed with low molar mass chitosan. A inverse behavior was observed when high oxidated CH was used.

Made available in DSpace on 2014-12-17T15:42:31Z (GMT). No. of bitstreams: 1 ClaudioLV.pdf: 1496908 bytes, checksum: 9da02c26d9351b21019488117fc27924 (MD5) Previous issue date: 2007-05-28 / Dispersions composed of polyelectrolyte complexes based on chitosan and poly(methacrylic acid), PMAA, were obtained by the dropping method and template polymerization. The effect of molecular weight of PMAA and ionic strength on the formation of chitosan/poly(methacrylic acid), CS/PMAA, complexes was evaluated using the dropping method. The increase in molecular weight of PMAA inhibited the formation of insoluble complexes, while the increase in ionic strength &#64257;rst favored the formation of the complex followed by inhibiting it at higher concentrations. The polyelectrolyte complexation was strongly dependent on macromolecular dimensions, both in terms of molecular weight and of coil expansion/contraction driven by polyelectrolyte effect. The resultant particles from dropping method and template polymerization were characterized as having regions with different charge densities: chitosan predominating in the core and poly(methacrylic acid) at the surface, the particles being negatively charged, as a consequence. Albumin was adsorbed on templatepolymerized CS/PMAA complexes (after crosslinking with glutardialdehyde) and pH was controlled in order to obtain two conditions: (i) adsorption of positively charged albumin, and (ii) adsorption of albumin at its isoelectric point. Adsorption isotherms and zeta potential measurements showed that albumin adsorption was controlled by hydrogen bonding/van der Waals interactions and that brushlike structures may enhance adsorption of albumin on these particles / Dispers?es formadas a partir de complexos polieletrol?ticos de quitosana e de poli(?cido metacr?lico), PMAA, foram obtidas tanto pelo m?todo de gotejamento, como pelo m?todo de polimeriza??o em molde. O efeito da massa molar do PMAA e da for?a i?nica na forma??o dos complexos de quitosana/poli(?cido metacr?lico), CS/PMAA, foi avaliado usando o m?todo de gotejamento. O aumento da massa molar do PMAA inibiu a forma??o dos complexos insol?veis, enquanto o aumento da for?a i?nica primeiramente favoreceu a forma??o dos complexos, depois a inibiu, em altas concentra??es de eletr?litos de baixa massa molar. A complexa??o dos polieletr?litos foi fortemente dependente das dimens?es macromoleculares, tanto em termos da massa molar quanto do efeito de expans?o/contra??o dos novelos, devido ao efeito polieletrol?tico. As part?culas resultantes tanto do m?todo de gotejamento, como da polimeriza??o em molde foram caracterizadas por apresentarem regi?es com diferentes densidades de carga: a quitosana predominantemente presente na regi?o central e o poli(?cido metacr?lico), na superf?cie, sendo, portanto, as part?culas carregadas negativamente. A albumina foi adsorvida nos complexos de CS/PMAA obtidos por polimeriza??o em molde (depois de sofrerem reticula??o covalente usando glutaralde?do) e o pH foi controlado a &#64257;m de se obter duas condi??es: (i) adsor??o de albumina carregada positivamente e (ii) adsor??o de albumina em seu ponto isoel?trico. As isotermas de adsor??o e as medidas de potencial zeta mostraram que a adsor??o da albumina foi controlada por liga??es de hidrog?nio/intera??es de van der Waals e que as estruturas em forma de escova puderam aumentar a adsor??o da albumina nessas part?culas

Yes / A new type of self-assembled polyelectrolyte complex nanocarrier composed of chondroitin (CHON) and protamine (PROT) was designed and the ability of the carriers to bind salmon calcitonin (sCT) was examined. The response of sCT-loaded CHON/PROT NPs to a change in the properties of the liquid medium, e.g. its pH, composition or ionic strength was studied and in vitro peptide release was assessed. The biocompatibility of the NPs was evaluated in Caco-2 cells. CHON/PROT NPs were successfully obtained with properties that were dependent on the concentration of the polyelectrolytes and their mixing ratio. X-ray diffraction determined the amorphous nature of the negatively charged NPs, while those with the positive surface potential were semi-crystalline. sCT was efficiently associated with the nanocarriers (98-100%) and a notably high drug loading (13-38%) was achieved. The particles had negative zeta potential values and were homogenously dispersed with sizes between 60 and 250 nm. CHON/PROT NPs released less than 10% of the total loaded peptide in the first hour of the in vitro release studies. The enthalpy of the decomposition exotherm correlated with the amount of sCT remaining in NPs after the release experiments. The composition of medium and its ionic strength was found to have a considerable influence on the release of sCT from CHON/PROT NPs. Complexation to CHON markedly reduced the toxic effects exerted by PROT and the NPs were compatible and well tolerated by Caco-2 cells.