L'objectif de cette thèse est d'évaluer le concept d'un dispositif intégré comprenant un bioréacteur à biofilm fluidisé bio-augmenté, couplé à une membrane de nanofiltration de type polyelectrolyte multicouche, destiné à éliminer les micropolluants en traitement tertiaire des eaux usées domestiques, traitées conventionnellement. Les résultats montrent que, pour des micropolluants ciblés, chacun des procédés est efficace comme traitement tertiaire. Les mécanismes biologiques et de rétention membranaires sont explicités. Cependant des challenges restent à relever en particulier pour l'étape de bio augmentation (survie et implantation de la souche apportée) pour une exploitation de cette étape. D'autre part, des investigations plus poussées sont nécessaires à l'élaboration d'une membrane fiable et robuste. Un tel procédé couplé MBBR-NF pourra alors être entièrement justifié dans le contexte d'une élimination performante de micropolluants ciblés. Il aura toute sa place dans le panel des technologies vertes pour la préservation de l'environnement. / This thesis aims at answering whether the concept of an integrated layout comprised of a coupled "bioaugmented moving bed biofilm reactors (bMBBRs) - polyelectrolyte multilayer (PEM)-based nanofiltration (NF) membrane" can be considered as a promising technology to eliminate target MPs from conventionally-treated municipal wastewater. Results presented herein indicate that each given component of the layout is efficient in the tertiary removal of MPs. Still, several challenges ahead of the process bioaugmentation (such as the survival and maintenance of inoculated strains) must be in-depth studied to find convenient operating solutions. On the other hand, further investigations are definitely needed to achieve a robust PEM-based membrane as a long-lasting technology. Even though a coupled bMBBR-NF system for enhanced MPs removal can be experimentally justified is, however, practically questionable. "The tale of bMBBR-NF" deserves much more scientific endeavors as plenty of environmental considerations are placed in, whereby achieving a future Green technology will not be far from our expectation.

Thesis advisor: Udayan Mohanty / The stabilization of particular conformations of protein and nucleic acid structure is believed to play an important role in many important biological functions. In chapter one, the α -helical conformation and structural stability of single and double stapled all- hydrocarbon cross-linked p53 peptides when bound and unbound to MDM2 are investigated. Our study provides a comprehensive rationalization of the relationship between peptide stapling strategy, the secondary structural stability, and the binding affinity of p53-MDM2 complex. In chapter two, we study counterion-mediated collapse of a strongly charged model polyelectrolyte chain by Group-II divalent metal cations using coarse-grained Brownian dynamics simulations. Polyelectrolyte effects govern the association of counterions with the chain. Large ions are less effective in counterion condensation than small ions. However, upon counterion condensation, the reduction of the backbone charge is independent of size of the metal cations. Above a threshold value of Coulomb strength parameter, counterion release entropy drives the formation of counterion-induced compact states. In chapter three, the nature of surface tension in the random first order theory of supercooled liquid is analyzed within the framework of Landau-Lifshitz fluctuation theory. We show that the surface tension of a droplet satisfies the differential equation 4πr2(dσ)+ 8πrσ(r)− Br1/2 = 0 , where B/ T = 12πkBcv , T is temperature, kB is dr Boltzmann constant, and cv is heat capacity. A consequence is that the slope of the relaxation time at the glass transition temperature, i.e., the fragility index, is expressed as the square of the ratio of heat capacity and configurational entropy of the supercooled liquid. When backbone extended nucleosides are incorporated into a double helix, a unique helical structure is formed. In chapter four, we find that the predicted stability of modified backbone DNA strands in aqueous solution is in good agreement with experimental melting temperature data. The incorporation of extended backbone nucleosides into a duplex results in elongation of the end-to-end chain distance due to the distortion of the B-DNA conformation at the mutated base-pair insertion. We also find that the modified backbone helical twist is approximately 40 degrees, larger than B-DNA helical twist and closer to the twist angle predicted for D-form DNA. The folding of RNA tertiary structure has been described as an equilibrium between partially folded I (intermediate) states, and the fully folded native conformation, or N state. RNA is highly sensitive to the ionic environment due to its negative charge, and tertiary structures tend to be strongly stabilized by Mg2+. There is a need for models capable of describing the ion atmosphere surrounding RNA with quantitative accuracy. In chapter 5, we present a generalized Manning condensation model of RNA electrostatics for studying the Mg2+-induced RNA folding of the 58mer ribosomal fragment. / Thesis (PhD) — Boston College, 2016. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Chemistry.

Polyelectrolyte microcapsules were first established in 1998 as a potential drug delivery vehicle. Despite being well-established, microcapsules have not yet been thoroughly considered as a viable means of targeted drug delivery. This is largely due to the fact that microcapsules are inherently prone to unspecific binding to cells and proteins. Targeted delivery of drugs to specific diseased sites in the body is an area of research that has attracted many studies, particularly in drug deliveries that utilise microparticles. By achieving targeted delivery of a drug, one can increase the efficacy of the treatment, thus, reducing unwanted side effects. This thesis investigates methods which can modify these microcapsules in order to fine tune the release of the encapsulated drug as well as site-specific delivery of these vesicles i.e. obtain spatiotemporal control. To this end, biodegradable microcapsules of varying constituents are manufactured and their biodegradability is indirectly measured through quantification of the release of an encapsulated fluorescent protein (Rhodamine B-BSA). Fluorometry analysis of the supernatants of these microcapsule suspensions indicated that microcapsules synthesised from poly-L-arginine and poly-L-glutamic acid have the ability to encapsulate bovine serum albumin (BSA) with a high encapsulation efficiency (79.7%). Furthermore, they are able to produce a sustained release of BSA over a period of 5 Days. To complement this controlled-release study, an investigation into self-degradable microcapsules was undertaken. To achieve this, proteinase was encapsulated in both biodegradable and non-biodegradable microcapsules of different thickness. Analysis of the protein release over a period of 24 hours revealed that the release profiles of these microcapsules can be successfully controlled. Biodegradable microcapsules released 87% more protein than their non-biodegradable counterpart after 2 hours of incubation in deionised water. This provides conclusive evidence that the biodegradable microcapsules were, indeed, self-degradable. The latter part of this thesis focuses on achieving specific and exclusive targeted delivery using polyelectrolyte microcapsules, with respect to protein substrates. This is accomplished by creating an antibody-functionalised poly(ethylene glycol) (PEG) assembly within the microcapsule structure. Site-specific adsorption of these microcapsules is tested using protein micropatterns. Results obtained from adsorption assays using anti-collagen type IV-functionalised microcapsules show a 600-fold increase in binding to collagen type IV islands, compared to control proteins (fibronectin and BSA). This proves that significant adsorption was achieved on the target protein, with unspecific adsorptions being heavily suppressed on control proteins. Furthermore, similar results were found when microcapsules were functionalised with anti-fibronectin and exposed to fibronectin, highlighting the versatility of this type of biofunctionalisation.

In this work we have explored the possibility of using surfactant/polyelectrolyte complexes as templates to synthesize inorganic mesostructured materials mainly with a film morphology. Inorganic species deposit in those regions of the films which are filled with a polymer hydrogel, surrounding the arrays of ordered surfactant micelles. This method produced thick robust films where the inorganic regions are reinforced and functionalized by the polymer, thus these materials are expected to have potential applications in separation, absorption, catalysis and chemical sensing. Initial work involved mixing silicate precursors directly into CTAB/PEI solutions to form highly ordered 2D hexagonal silica films at the air/water interface. Time resolved synchrotron SAXS allowed investigation of processes occurring in solution during the reaction, from which a film growth mechanism was proposed. Films had good thermal properties and after post-synthesis TMOS vapour treatment, retained structure upon template removal. Silica gel monoliths with various mesostructures were also rapidly fabricated in one minute with surfactant/LPEI complexes. Cat-ionic surfactant mixtures with polymers were also employed to template silica films with different cubic mesostructures at the air/water interface. The mesophases of the interfacial films were enriched due to more complicated interactions between the polymers and two surfactants. Polymer molecular weight, total composite concentration, chemical nature of the polymer as well as the cationic-anionic surfactant molar ratio was used to systematically control the silica film mesophase. Robust titania films were also prepared with cat-anionic surfactant mixtures and polymers at the air/water interface. Although the film mesostructure was lost after calcination, the in-situ and dry free-standing films display ordered cubic mesostructures and the films are stable to calcination after post-synthesis treatment. Ordered lamellar iron oxide films templated by SDS/LPEI complexes were also prepared at the air/water interface.

This thesis reports an investigation from the “bedside” back to the “bench”. That is, from the economic evaluation of a medical intervention to basic research and development of a contractility assay. The underlying theme of this thesis is cellular contractility, which was stimulated from our laboratory’s work in the microvascular complications of Diabetic Retinopathy (DR). The health economic perspective of this thesis evaluates the cost effectiveness and cost utility of DR prognosis using the prog-DR test. This novel prognostic test developed in our laboratory relies on the contractile response of blood vessels to detect subjects with high risk of developing DR. Markov modeling based on information in the literature was used to estimate the outcomes of a hypothetical population. The costs, health and utility outcomes of DR were compared to the potential outcomes if the prog-DR test was used. The model show that the prog-DR test can improve the health of the hypothetical population as measured in the number of life years (LY), sight years (SY) and quality-adjusted life years (QALY). The prog-DR test was more cost effective than the benchmark of annual or bi-annual screening and the incremental cost effectiveness ratio (ICER) appears to be at an acceptable level. Scenario and sensitivity analysis also show that the cost effectiveness of the prog-DR test can be improved by (i) better blood glucose management post prog-DR test, (ii) targeted screening (as opposed to population-wide screening) and (iii) reduced costs of both screening and management of DM and DR. The physiological perspective of the thesis aimed to develop a contractility assay for DR that was based on a 3D scaffold, which was affordable, easy to make and mimicked the three dimensional physiological environment of blood vessels. The contractility assay was developed using a 3D, hollow scaffold (PE-PAH capsule) and involved (i) the selection of the optimal core material, (ii) optimisation of the manufacturing process, (iii) characterisation of the scaffold and (iv) ensuring that cells can be grown on it. The cyto-biocompatibility of the candidate polyelectrolyte Poly(Sodium 4-Styrene Sulfonate) (PSS) and Poly(Allylamine Hydrochloride) (PAH) in the thin films format were investigated using three different cell lines and the effects of these thin films were also compared to titanium and titanium nitride thin films. In essence, PSS and PAH are not cytotoxic and was used to develop the contractile scaffold, PE-PAH capsule. This scaffold is relative elastic and the contractile force exerted by the 3T3-L1 cells was calculated based on the deformation of the PE-PAH capsule. The contractility assay was sufficiently sensitive to detect the nano-Newton magnitude of force developed by individual cells and discriminated the change in force due to disruption of the F-actin cytoskeleton by forskolin and cytochalasin D.

The formation of colloids by the controlled reduction, nucleation, and growth of inorganic precursor salts in different media has been investigated for more than a century. Recently, the preparation of ultrafine particles has received much attention since they can offer highly promising and novel options for a wide range of technical applications (nanotechnology, electrooptical devices, pharmaceutics, etc). The interest derives from the well-known fact that properties of advanced materials are critically dependent on the microstructure of the sample. Control of size, size distribution and morphology of the individual grains or crystallites is of the utmost importance in order to obtain the material characteristics desired. Several methods can be employed for the synthesis of nanoparticles. On the one hand, the reduction can occur in diluted aqueous or alcoholic solutions. On the other hand, the reduction process can be realized in a template phase, e.g. in well-defined microemulsion droplets. However, the stability of the nanoparticles formed mainly depends on their surface charge and it can be influenced with some added protective components. Quite different types of polymers, including polyelectrolytes and amphiphilic block copolymers, can for instance be used as protecting agents. The reduction and stabilization of metal colloids in aqueous solution by adding self-synthesized hydrophobically modified polyelectrolytes were studied in much more details. The polymers used are hydrophobically modified derivatives of poly(sodium acrylate) and of maleamic acid copolymers as well as the commercially available branched poly(ethyleneimine). The first notable result is that the polyelectrolytes used can act alone as both reducing and stabilizing agent for the preparation of gold nanoparticles. The investigation was then focused on the influence of the hydrophobic substitution of the polymer backbone on the reduction and stabilization processes. First of all, the polymers were added at room temperature and the reduction process was investigated over a longer time period (up to 8 days). In comparison, the reduction process was realized faster at higher temperature, i.e. 100°C. In both cases metal nanoparticles of colloidal dimensions can be produced. However, the size and shape of the individual nanoparticles mainly depends on the polymer added and the temperature procedure used. In a second part, the influence of the prior mentioned polyelectrolytes was investigated on the phase behaviour as well as on the properties of the inverse micellar region (L2 phase) of quaternary systems consisting of a surfactant, toluene-pentanol (1:1) and water. The majority of the present work has been made with the anionic surfactant sodium dodecylsulfate (SDS) and the cationic surfactant cetyltrimethylammonium bromide (CTAB) since they can interact with the oppositely charged polyelectrolytes and the microemulsions formed using these surfactants present a large water-in-oil region. Subsequently, the polymer-modified microemulsions were used as new templates for the synthesis of inorganic particles, ranging from metals to complex crystallites, of very small size. The water droplets can indeed act as nanoreactors for the nucleation and growth of the particles, and the added polymer can influence the droplet size, the droplet-droplet interactions, as well as the stability of the surfactant film by the formation of polymer-surfactant complexes. One further advantage of the polymer-modified microemulsions is the possibility to stabilize the primary formed nanoparticles via a polymer adsorption (steric and/or electrostatic stabilization). Thus, the polyelectrolyte-modified nanoparticles formed can be redispersed without flocculation after solvent evaporation. / Die Bildung von Kolloiden durch kontrollierte Reduktion, durch Keimbildung und durch Wachstum anorganischer Precurser in unterschiedlichen Medien wird seit mehr als einem Jahrhundert intensiv beforscht. Vor kurzem hat die Herstellung ultrafeiner Partikel viel Aufmerksamkeit errungen, da sich hieraus vielversprechende neue Möglichkeiten für ein breites Spektrum an technischen Anwendungen (Nanotechnologie, elektrooptische Materialen, Pharmazeutik, usw.) ergeben. Das Interesse leitet sich von der weithin bekannten Tatsache ab, dass die Eigenschaften der „advanced materials“ von der Mikrostruktur der Probe deutlich abhängig sind. Die gezielte Steuerung der Größe, der Größenverteilung und der Morphologie der einzelnen Keime oder Kristallite ist von größter Wichtigkeit, um die gewünschten Eigenschaften zu erreichen. Verschiedene Methoden können für die Synthese von Nanopartikel verwendet werden. Einerseits kann eine Reduktion in verdünnten wässrigen oder alkoholischen Lösungen stattfinden, andererseits kann der Reduktionsprozess in einer Templatphase, z.B. in definierten Mikroemulsionströpfchen stattfinden. Die Stabilität der produzierten Nanopartikel hängt hauptsächlich von ihrer Oberflächenladung ab, welche durch schützende Komponenten zusätzlich beeinflusst werden kann. Verschiedene Arten von Polymeren, einschließlich Polyelektrolyte und amphiphile Blockcopolymere, können als solche Komponenten benutzt werden. Die Reduktion und Stabilisierung von Metallkolloiden in der wässrigen Lösung durch Addition von hydrophob modifizierten Polyelektrolyten werden bereits ausführlich studiert. Die verwendeten Polymere sind hydrophob modifizierte Derivate des Natrium-Polyacrylat, der Maleinsäure Copolymere sowie das verzweigte Poly(ethylenimin). Erstaunlicherweise genügt bereits die Anwesenheit die verwendeten Polyelektrolyte zu Reduzierung und Stabilisierung der Goldnanopartikel. Darüber hinaus wurde der Einfluss der hydrophoben Seitenkette des Polymer auf den Reduktions- und Stabilisierungsprozess bei unterschiedliche Reaktionstemperatur untersucht. In beiden Fällen können Metallnanopartikel kolloidaler Größe hergestellt werden, jedoch hängt die Größe und die Form der einzelnen Nanopartikel hauptsächlich vom dem zugefügten Polymer und vom verwendeten Temperaturverfahren ab. Im zweiten Teil wurde der Einfluss der vorher erwähnten Polyelektrolyte auf das Phaseverhalten sowie auf die Eigenschaften der inversen mizellaren Region (L2 Phase) der quaternären Systeme untersucht, die aus einem Tensid, Toluol-Pentanol – Gemisch (1:1) sowie Wasser bestehen. Dabei wurden hauptsächlich ionische Tenside, wie z.B. das anionische Natriumdodecylsulfate (SDS) und das kationische Cetyltrimethylammonium-bromid (CTAB) verwendet, da sie mit den geladenen Polyelektrolyten wechselwirken können. Darüber hinaus wurden die polymer-modifizierten Mikroemulsionen als neue Template für die Synthese von anorganischen Nanopartikeln verwendet. Die Wassertröpfchen können in der Tat als Nanoreaktoren für die Keimbildung und das Wachstum der Partikel dienen, und das zugefügte Polymer kann die Tröpfchengröße, die Tröpfchen-Tröpfchen Wechselwirkungen, sowie die Stabilität des Tensidfilms durch Polyelektrolyt-Tensid Komplexbildung beeinflussen. Ein weiterer Vorteil der polymer-modifizierten Mikroemulsionen ist die Stabilizierung der produzierten Primärteilchen über eine Polymeradsorption (durch sterische bzw. elektrostatische Stabilisierung), welche eine Redispergierung der Polyelektrolyte-modifiziert Nanopartikel, nach Lösungsmittel-verdampfung, ohne Aggregation der Partikel erlaubt.

Polyelectrolyte microcapsules containing stimuli-responsive polymers have potential applications in the fields of sensors or actuators, stimulable microcontainers and controlled drug delivery. Such capsules were prepared, with the focus on pH-sensitivity and carbohydrate-sensing. First, pH-responsive polyelectrolyte capsules were produced by means of electrostatic layer-by-layer assembly of oppositely charged weak polyelectrolytes onto colloidal templates that were subsequently removed. The capsules were composed of poly(allylamine hydrochloride) (PAH) and poly(methacrylic acid) (PMA) or poly(4-vinylpyridine) (P4VP) and PMA and varied considerably in their hydrophobicity and the influence of secondary interactions. These polymers were assembled onto CaCO3 and SiO2 particles with diameters of ~ 5 µm, and a new method for the removal of the silica template under mild conditions was proposed. The pH-dependent stability of PAH/PMA and P4VP/PMA capsules was studied by confocal laser scanning microscopy (CLSM). They were stable over a wide pH-range and exhibited a pronounced swelling at the edges of stability, which was attributed to uncompensated positive or negative charges within the multilayers. The swollen state could be stabilized when the electrostatic repulsion was counteracted by hydrogen-bonding, hydrophobic interactions or polymeric entanglement. This stabilization made it possible to reversibly swell and shrink the capsules by tuning the pH of the solution. The pH-dependent ionization degree of PMA was used to modulate the binding of calcium ions. In addition to the pH-sensitivity, the stability and the swelling degree of these capsules at a given pH could be modified, when the ionic strength of the medium was altered. The reversible swelling was accompanied by reversible permeability changes for low and high molecular weight substances. The permeability for glucose was evaluated by studying the time-dependence of the buckling of the capsule walls in glucose solutions and the reversible permeability modulation was used for the encapsulation of polymeric material. A theoretical model was proposed to explain the pH-dependent size variations that took into account an osmotic expanding force and an elastic restoring force to evaluate the pH-dependent size changes of weak polyelectrolyte capsules. Second, sugar-sensitive multilayers were assembled using the reversible covalent ester formation between the polysaccharide mannan and phenylboronic acid moieties that were grafted onto poly(acrylic acid) (PAA). The resulting multilayer films were sensitive to several carbohydrates, showing the highest sensitivity to fructose. The response to carbohydrates resulted from the competitive binding of small molecular weight sugars and mannan to the boronic acid groups within the film, and was observed as a fast dissolution of the multilayers, when they were brought into contact with the sugar-containing solution above a critical concentration. It was also possible to prepare carbohydrate-sensitive multilayer capsules, and their sugar-dependent stability was investigated by following the release of encapsulated rhodamine-labeled bovine serum albumin (TRITC-BSA). / Die Entwicklung von stimulisensitiven Systemen hat in den vergangenen Jahren großes Interesse hervorgerufen. Diese Systeme verändern ihre Eigenschaften in Abhängigkeit von äußeren Einflüssen und haben potenzielle Anwendungsgebiete im Bereich von Sensoren und der kontrollierten Wirkstofffreisetzung. Beispiele für äußere Einflüsse sind die Temperatur, die Salzkonzentration der Lösung, der pH-Wert oder die Gegenwart bestimmter chemischer Substanzen. Polyelektrolyt-Multischicht-Hohlkugeln stellen viel versprechende stimulisensitive Systeme dar, da sie aus dünnen Membranen mit veränderbaren Eigenschaften aufgebaut sind, und eingekapselte Substanzen definiert freigesetzt werden können. Im Rahmen dieser Arbeit wurden zum einen Polyelektrolytkapseln untersucht, deren Stabilität, Größe und Durchlässigkeit abhängig ist vom pH-Wert und der Salzkonzentration der Lösung. Zum anderen wurden Hohlkugeln entwickelt, deren Stabilität durch die Gegenwart verschiedener Zucker in der umgebenden Lösung beeinflusst wurde. Hierbei war es möglich, eine eingekapselte Substanz in Abhängigkeit der Zuckerkonzentration der Lösung freizusetzen. pH-sensitive Polyelektrolyt-Hohlkugeln wurden mit Hilfe der elektrostatischen Layer-by-Layer Methode aufgebaut. Hierbei wurden abwechselnd schwache Polysäuren und Polybasen auf kolloidalen Partikeln adsorbiert und in einem weiteren Schritt wurde das Templat entfernt. Als Polyelektrolyte fanden Poly(allylaminhydrochlorid) (PAH) und Poly(methacrylsäure) (PMA) bzw. Poly(4-vinylpyridin) (P4VP) und PMA Verwendung. Diese zwei Systeme unterscheiden sich in ihren hydrophoben Eigenschaften und dem Einfluss von Sekundärwechselwirkungen. Die Polymere wurden auf CaCO3 und SiO2 Partikeln mit Durchmessern von ~ 5 µm aufgebracht, und eine neue Methode zum Entfernen des Silica-Templats wurde vorgestellt. Die pH-abhängige Stabilität von PAH/PMA und P4VP/PMA Hohlkugeln wurde mit der konfokalen Fluoreszenzmikroskopie untersucht. Beide Systeme waren über einen breiten pH-Bereich stabil und zeigten an der Grenze der Stabilität ein ausgeprägtes Schwellen, das den unkompensierten positiven oder negativen Ladungen der Mulitschichten zugeordnet werden konnte. Gab es innerhalb der Multischichten stabilisierende Wechselwirkungen, z.B. Wasserstoffbrückenbindungen oder hydrophobe Anziehungskräfte, welche der elektrostatischen Destabilisierung entgegenwirkten, so ließ sich der geschwollene Zustand stabilisieren. Diese Stabilisierung ermöglichte das reversible Schwellen der Hohlkugeln durch Veränderung des pH-Wertes der Lösung. Der pH-abhängige Ionisationsgrad von PMA wurde zur Bindung von Calcium-Ionen verwendet. Die Stabilität und das Schwellen der Hohlkugeln konnten durch Variation der Ionenstärke der Lösung modifiziert werden. Das reversible Schwellen wurde von reversiblen Permeabilitätsänderungen für nieder- und hochmolekulare Substanzen begleitet. Die Permeabilität für Glucose wurde mit Hilfe der Zeitabhängigkeit der Verformung der Kapselmembranen in Glucoselösungen untersucht, und die reversible Veränderung der Permeabilität wurde zur Einkapselung von Polymeren verwendet. Die pH-abhängigen Größenänderungen ließen sich mit einem theoretischen Modell beschreiben, das von einem Kräftegleichgewicht zwischen einer osmotischen expansiven Kraft und einer elastischen kontraktiven Kraft ausging. Als zweites stimulisensitives System wurden zuckersensitive Multischichten untersucht. Diese wurden über reversible kovalente Esterbindungen stabilisiert. Als Polymere für den Schichtaufbau fanden das Polysaccharid Mannan und Polyacrylsäure, die mit Aminophenylboronsäure modifiziert war, Verwendung. Die daraus entstehenden Multischichten waren sensitiv gegenüber verschiedenen Kohlenhydraten, mit der größten Selektivität für Fructose. Diese Affinitäten ergaben sich aus der kompetitiven Bindung zwischen niedermolekularen Zuckern und Mannan an die Boronsäuregruppen. Oberhalb einer kritischen Zuckerkonzentration kam es zur schnellen Auflösung der Multischichten. Darüber hinaus war es möglich zuckersensitive Polymerkapseln herzustellen, deren zuckerabhängige Stabilität durch die Freisetzung von eingekapseltem Rinderserumalbumin verfolgt wurde.

Contact-active antibacterial surfaces with irreversibly attached antibacterial com-pounds are a sustainable alternative to traditional biocides. No chemicals are released into nature and the antibacterial mechanism reduces the risk of the evolution of re-sistant bacteria. However, the preparation of such surfaces is far from sustainable, as organic solvents and harsh reaction conditions commonly are required. An alter-native option is to use polyelectrolyte multilayers (PEM), based on physical ad-sorption, which can be performed in water-based solutions at room temperature. Although contact-active antibacterial PEMs have been reported previously, this is the first study of renewable cellulosic wood fibres. The build-up of cationic polymer polyvinylamine (PVAm) and anionic polyacrylic acid (PAA) multilayers on model surfaces was studied to optimise adsorption. The amount of adsorbed polyelectrolytes was continuously growing with increasing number of layers, but remained dense and flat as the number of layers increased. The largest adsorption was obtained at a high salt concentration, which shielded the repulsion between the polymers. Model surfaces were also used to evaluate the influence of the polymer and number of layers on the antibacterial properties. Multilayers on model surfaces showed a low bacteriostatic effect, with up to approximately 40 % inhibition for 3 layers of un-modified PVAm/PAA. In contrast, when the same multilayers were applied on cel-lulosic fibres, bacterial-growth inhibition of > 99.9% was obtained. Hydrophobically modified PVAm did not yield better results, despite being superior in solution. An increase in fibre charge by fibre oxidation led to the largest amount of adsorbed pol-ymer and the best antibacterial properties, an effect that lasted for weeks. Electron microscopy study of bacteria on the fibres showed that the bacteria interacted more on a highly charged surface and that the morphology of the bacterial cell could be affected. The effect was suggested to be due to electrostatic interaction with the pos-itively charged modified fibres. The promising results offer the possibilities of a new generation of antibacterial surfaces based on a renewable resource. / Antibakteriella kontaktaktiva ytor som har ett antibakteriellt ämne permanent fäst på ytan är ett miljövänligt och säkrare alternativ till traditionell biocidanvändning. Inget utsläpp av giftiga ämnen sker från ytorna och detta tillsammans med den anti-bakteriella mekanismen minskar risken för att bakterierna utvecklar resistens. Till-verkningsprocessen i sig har dock hittills varit allt annat än miljövänlig, då den ke-miska modifieringen kräver organiska lösningsmedel och har reaktioner som kräver speciella villkor, t ex höga temperaturer. En alternativ ytmodifiering är att använda sig av fysikalisk adsorption av polyelektrolyter i multiskikt, eftersom detta kan göras i vat-tenlösningar och i rumstemperatur. Det här arbetet är det första som beskriver kon-takt-aktiva multilager på förnyelsebara svedbaserade cellulosafiber. Som ett första steg gjordes en adsorptionsstudie på modellytor för att optimera ad-sorptionen av katjonisk polyvinylamin (PVAm) och anjonisk polyakrylsyra (PAA). Med ökande antal lager ökade totala mängden adsorberad polymer samtidigt som multilagerna förblev platta och täta. Den högsta adsorptionen skedde vid en hög salt-halt som minimerade den elektrostatiska repulsionen mellan polymerkedjorna. Modellytor användes även för att studera hur de antibakteriella egenskaperna påver-kades av polymermodifiering och av antal lager. På dessa ytor uppmättes en låg bakte-riostatisk effekt med upp till 40 % inhibering av bakterietillväxten för tre lager av PVAm./PAA När däremot samma multilager fanns på cellulosafiber ökade in-hiberingen till uppemot 99.9 %. Hydrofobmodifiering av PVAm påverkade inte det antibakteriella resultatet när de var i multilager, trots bevisad ökad verkan i lösning. Genom att via oxidering öka fiberladdningen kunde mängden adsorberad polymer yt-terligare öka och resulterade i en förbättrad antibakteriell verkan som höll i sig i flera veckor. Elektronmikroskopi av bakterier på fiber visade en ökad interaktion med hög-laddade ytor och att bakteriernas cellmorfologi kan påverkas av ytorna.Den observerade antibakteriella effekten föreslås vara en följd av elektrostatisk inter-aktion mellan de negativt laddade bakterierna och positivt laddade modifierade fibrena. Resultaten är lovande och banar väg för nya kontakt-aktiva antibakteriella material. / Biointeractive fibres with antibacterial properties

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Conjugated polymers have been found useful in a wide range of applications such as solar cells, sensor elements and printed electronics, due to their optical and electronic properties. Functionalization with charged side chains has enabled water solubility, resulting in an enhanced interaction with biomolecules. This thesis focus on the emerging research fields, where these conjugated polyelectrolytes (CPEs) are combined with biomolecules for biological sensing and DNA nanowire assembling. CPEs have shown large potential in biomolecular detection where the optical read out is due to the geometrical alternation in the backbone and aggregation state. This thesis focused on transferring the biomolecular detection to a surface of CPEs. The characterization of the CPE layer show that a hydrogel can be formed, and how the layer can undergo geometrical changes upon external stimulus such as pH change. A selective sensor surface can be created by imprinting ssDNA or an antibody in the CPE layer. The discrimination for complementary DNA hybridization and specific antibody interaction can be monitored by surface plasmon resonance or quartz crystal microbalance. We have also taken the step out from the controlled test tube experiments to the complex environment of the cell showing the potential for staining of compartments and structures in live and fixed cell. Depending on the conditions and CPE used, cell nuclei, acidic vesicles and cytoskeleton structure can be visualized. Furthermore, the live staining shows no sign of toxic effect on cultured fibroblasts. CPEs can also be a valuable element when assembling electronics in the true nano regime. I have used DNA as building template due to its attractive size features, with a width of around 2 nm and a length scale in the µm regime, and the inbuilt base-paring recognition elements. This thesis shows how DNA can be decorated with CPEs and stretched on surfaces into a model for aligned semiconducting nanowire geometries. Not only making the template structures is of importance, but also how to place them on the correct surface position, i.e. on electrodes. Strategies for positioning DNA nanowires using transfer printing and surface energy patterning methods have therefore been developed in the thesis. The stretched DNA decorated with CPE also offers a way to further study the molecular binding interaction between the two molecules. Single molecular spectroscopy in combination with polarization has given information of the variation of the CPE binding along a DNA chain.