No description available.

We report the design and assembly of three-dimensional (3D) multi-cellular liver models comprised of primary rat hepatocytes, liver sinusoidal endothelial cells (LSECs), and Kupffer cells (KCs). LSECs and KCs in the liver model were separated from hepatocytes by a nanoscale, detachable, optically transparent chitosan and hyaluronic acid (HA) polyelectrolyte multilayer (PEM) film. The properties of the PEM were tuned to mimic the Space of Disse found in liver. The thickness of the detachable PEM was 650 to 1000 nm under hydrated conditions. The Young's modulus of the PEM was approximately 42 kPa, well within the range of modulus values reported for bulk liver. The 3D liver models comprised of all three cell types and a detachable PEM exhibited stable urea production and increased albumin secretion over a 12 day culture period. Additionally, the 3D liver model maintained the phenotype of both LSECs and KCs over the 12 day culture period, verified by CD32b and CD163 staining, respectively. Additionally, CYP1A1 enzyme activity increased significantly in the 3D liver models. The number of hepatocytes in the 3D liver model increased by approximately 60% on day 16 of culture compared to day 4 indicating. Furthermore, only the 3D hepatic model maintained cellular compositions virtually identical to those found in vivo. DNA microarray measurements were conducted on the hepatocyte fractions of the 3D liver mimic to obtain insights into hepatic processes. Gene sets up-regulated in the 3D liver model were related to proliferation, migration, and deposition of extracellular matrix, all functions observed in regenerating hepatocytes. Taken together, these results suggest that inter-cellular signaling between the different cell types in the 3D liver model led to increased hepatic functions. To the best of our knowledge, this is the first study where three of the major hepatic cell types have been incorporated into a model that closely mimics the structure of the sinusoid. These studies demonstrate that the multi-cellular liver models are physiologically relevant. Such models are very promising to conduct detailed investigations into hepatic inter-cellular signaling. / Ph. D.

<p>In recent years, antibacterial surfaces have been a subject of increased interest. Especiallyinteresting are non-leaching, contact-active surfaces that physically disrupts the bacterialcell using immobilised cationic polymers. Thus the risks of bacterial resistance and discharge of hazardous biocides is minimised. The assembly of such surfaces is elaborate andusually involves organic solvents. Here, polyelectrolyte multilayers (PEM) are proposed as an effective surface modification method, with an overall goal of producing antibacterial cellulose fibres. The PEM process is based on physical adsorption of oppositely charged polymers in aqueous solutions. Multilayers were formed with the bactericidal polymer polyvinylamine (PVAm) and polyacrylic acid. PVAm compounds with hydrophobic modificationswere applied as well, as they possess increased antibacterial activity in solution.</p><p>In this work, the multilayer formation was studied on model surfaces of silicone oxide and glass in order to obtain fundamental knowledge of the polymer system. QCM-D and reflectometry, which detect total mass including bound water and polymer mass only, respectively, were used to analyse the layer formation. Salt-concentrations were varied at 1, 10 or 100 mM NaCl. A stepwise multilayer formation with exponential-like polymer adsorption but with decreasing water content for each layer was seen at all salt concentrations.A higher salt concentration resulted in an increased adsorbed mass. No significant differences in adsorption between the modified and unmodified PVAm could be detected. AFM imaging applied to multilayers having nine layers showed large surface aggregates under high salt conditions for the C6-modified PVAm. Dynamic light scattering showed that the polymer occurred as single molecules in solution; hence it was concluded that theaggregation is surface-associated.</p><p>The multilayers were then tested for bacterial growth inhibition. The relative bacterial inhibition was time-dependent, as the surface was saturated with bacteria over time. After two hours, a maximal inhibition of 99 % could be observed for the multilayers. After eight hours, a moderate inhibition of less than 40 % was detected. Using multilayers affected the results positively compared to single layers. After three layers, though, no further reductionwas seen. Viability staining of the surface-adhered bacteria revealed that the adhered bacteria had intact membranes. Therefore, the microbiological properties of the multilayers can at this point be described more as growth-inhibiting by bacterial adhesion effectsthan as biocidal. However, this work has shown the importance of combining surface characterisation and microbial testing to understand the bacteria-surface interaction.</p> / Biointeractive fibres

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 &gt; 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

A major challenge in hepatic tissue engineering is that liver cells rapidly lose their phenotype in in vitro cell culture systems. For this reason, it is necessary to design biomaterials that can support and enhance hepatic functions. Hepatocytes have a surface protein, called the asialoglycoprotein receptor (ASGP-R), which interacts with galactose via a specific receptor-ligand interaction. Polyelectrolyte multilayers (PEMs) were prepared by the layer by layer method, which is based on electrostatic attractions between oppositely charged polyelectrolytes (PEs). Anionic (hyaluronic acid) and cationic (chitosan and galactosylated chitosan) PEs were used in the fabrication of detachable, free-standing PEMs. The main focus of this study is the design of PEMs comprised of 50 bilayers of PEs. PEMs that contained galactose functional groups were assembled with either 5 or 10 bilayers of galactosylated chitosan (5 - 10 % of galactosylation). Optical properties, solvent stability and surface topography of the PEMs were measured. / Master of Science

"Polyelectrolyte multilayer (PEM) thin films prepared via the Layer-by-Layer (LbL) deposition technique are of special interest in this research. The purpose of this study is to replace current mechanical closure systems, based on hook-and-loop type fasteners (i.e. Velcro), with PEM thin film systems. The technique is simple, cheap, versatile and environmental friendly; as a consequence a variety of thin films can be easily fabricated. By proposing PEMs as non-mechanical and nanoscopic molecular closures, we aim to obtain hermetic sealing, good adhesive strength, and peel off ease. Atomic force microscopy (AFM) and colloidal probe techniques were used to characterize the morphology, roughness and adhesive properties of PEMs. AFM measurements were conducted in air, necessarily requiring careful control of ambient humidity. PEMs were formed by consecutive deposition of polyanions and polycations on a charged polyethylene terephthalate (PET) solid surface, the result of which was stable nanostructured films. By systemically varying the parameters of PEM build-up process: different combinations of polyelectrolytes, different numbers of bilayers (polyanion/polycation pairs), and miscellaneous types and concentrations of salts (NaCl, NaBr and NaF salts at 0.5 M and 1.0 M concentrations), the adhesion and morphology of PEMs were thoroughly investigated. The PEM thin films specifically investigated include poly(ethyleneimine) (PEI), poly(styrene sulfonate) (PSS), poly(allylamine hydrochloride) (PAH), poly(acrylic acid) (PAA), and poly(diallydimethylammonium chloride) (PDADMAC). Silica colloidal probes were utilized in the investigation, some of which were functionalized with COOH and/or coated with PEI-PSS. Silica colloidal probes were used in order to quantify interaction forces on the PEMs. A functionalized silica colloidal probe (a probe with COOH surface chemistry) and a silica colloidal probe coated with PEI-PSS were used to simulate PEM-PEM interactions. The results suggest that adhesion in the PEMs depend on the number of layers, the salt concentration and the salt type used during the build-up process, the environmental conditions where the adhesion force measurements were made, and the choice of probe. "

Die Diffusion durch Polyelektrolytschichten ist bei vielen biotechnologisch-pharmazeutischen Anwendungen im Nanometerbereich von Bedeutung. Bei Experimenten wurde gefunden, dass bei der Diffusion eines Quenchers in mit Fluoreszenzfarbstoff markierten Polyelektrolytmultischichten die Kinetik des Diffusionsprozesses Charakteristika einer anormalen Diffusion aufweist. In dieser Arbeit wird qualitativ und quantitativ gezeigt, dass die Diffusion des Quenchers mit Subdiffusion beschrieben werden kann. Der gemessene Diffusionsprozess kann durch eine Superposition von Markov‘schen Diffusionsprozessen dargestellt werden. Das wird mit einer Monte-Carlo-Simulation nachgewiesen, die auf der analytischen Lösung des vorliegenden Reaktions-Diffusionsgleichungssystems basiert. Die experimentellen Daten werden im Zusammenhang mit der strukturellen Basis der parallel ablaufenden Diffusionsprozesse diskutiert.

No description available.

In recent years, antibacterial surfaces have been a subject of increased interest. Especiallyinteresting are non-leaching, contact-active surfaces that physically disrupts the bacterialcell using immobilised cationic polymers. Thus the risks of bacterial resistance and discharge of hazardous biocides is minimised. The assembly of such surfaces is elaborate andusually involves organic solvents. Here, polyelectrolyte multilayers (PEM) are proposed as an effective surface modification method, with an overall goal of producing antibacterial cellulose fibres. The PEM process is based on physical adsorption of oppositely charged polymers in aqueous solutions. Multilayers were formed with the bactericidal polymer polyvinylamine (PVAm) and polyacrylic acid. PVAm compounds with hydrophobic modificationswere applied as well, as they possess increased antibacterial activity in solution. In this work, the multilayer formation was studied on model surfaces of silicone oxide and glass in order to obtain fundamental knowledge of the polymer system. QCM-D and reflectometry, which detect total mass including bound water and polymer mass only, respectively, were used to analyse the layer formation. Salt-concentrations were varied at 1, 10 or 100 mM NaCl. A stepwise multilayer formation with exponential-like polymer adsorption but with decreasing water content for each layer was seen at all salt concentrations.A higher salt concentration resulted in an increased adsorbed mass. No significant differences in adsorption between the modified and unmodified PVAm could be detected. AFM imaging applied to multilayers having nine layers showed large surface aggregates under high salt conditions for the C6-modified PVAm. Dynamic light scattering showed that the polymer occurred as single molecules in solution; hence it was concluded that theaggregation is surface-associated. The multilayers were then tested for bacterial growth inhibition. The relative bacterial inhibition was time-dependent, as the surface was saturated with bacteria over time. After two hours, a maximal inhibition of 99 % could be observed for the multilayers. After eight hours, a moderate inhibition of less than 40 % was detected. Using multilayers affected the results positively compared to single layers. After three layers, though, no further reductionwas seen. Viability staining of the surface-adhered bacteria revealed that the adhered bacteria had intact membranes. Therefore, the microbiological properties of the multilayers can at this point be described more as growth-inhibiting by bacterial adhesion effectsthan as biocidal. However, this work has shown the importance of combining surface characterisation and microbial testing to understand the bacteria-surface interaction. / Biointeractive fibres

Integration of living cells with novel microdevices requires the development of innovative technologies for manipulating cells. Chemical surface patterning has been proven as an effective method to control the attachment and growth of diverse cell populations. Patterning polyelectrolyte multilayers through the combination of layer-by-layer self-assembly technique and photolithography offers a simple, versatile and silicon compatible approach that overcomes chemical surface patterning limitations, such as short-term stability and low protein adsorption resistance. In this study, direct photolithographic patterning of PAA/PAAm and PAA/PAH polyelectrolyte multilayers was developed to pattern mammalian neuronal, skeletal and cardiac muscle cells. For all studied cell types, PAA/PAAm multilayers behaved as a negative surface, completely preventing cell attachment. In contrast, PAA/PAH multilayers have shown a cell-selective behavior, promoting the attachment and growth of neuronal cells (embryonic rat hippocampal and NG108-15 cells) to a greater extent, while providing a little attachment for neonatal rat cardiac and skeletal muscle cells (C2C12 cell line). PAA/PAAm multilayer cellular patterns have also shown a remarkable protein adsorption resistance. Protein adsorption protocols commonly used for surface treatment in cell culture did not compromise the cell attachment inhibiting feature of the PAA/PAAm multilayer patterns. The combination of polyelectrolyte multilayer patterns with different adsorbed proteins could expand the applicability of this technology to cell types that require specific proteins either on the surface or in the medium for attachment or differentiation, and could not be patterned using the traditional methods.