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Biodistribution of nanoparticle-based intracellular delivery systems is mediated primarily by particle size and physicochemical properties. As such, overcoming the rapid removal of these by the reticuloendothelial system remains a significant challenge. To date, a number of copolymer nanoparticle systems based on 2-methacryloyloxyethyl phosphorylcholine (MPC) with 2-(diisopropylamino)ethyl methacrylate (DPA), displaying biomimetic and pH responsive properties, have been published, however these have been predominately polymersome based, whilst micelle systems have remained relatively unexplored. This study utilised nanoprecipitation to investigate the effects of solvent and buffer choice upon micelle size and polydispersity, and found using methanol produced monodisperse micelles of circa 70 nm diameter, whilst ethanol produced polydisperse systems with nanoparticles of circa 128 nm diameter. The choice of aqueous buffer, dialysis of the systems, extended storage, and exposure to a wide temperature range (5-70 °C) had no significant effect on micelle size, and the systems were highly resistant to dilution, indicating excellent colloidal stability. Optimisation of the nanoprecipitation process, post precipitation, was investigated, and model drugs successfully loaded whilst maintaining system stability. Subsequent in vitro studies suggested that the micelles were of negligible cellular toxicity, and an apparent cellular uptake was observed via confocal laser scanning microscopy. This paper presents the first report of an optimised nanoprecipitation methodology for the formation of MPC-DPA nanoparticle micelles, and in doing so achieved monodisperse systems with the size and physicochemical characteristics seen as desirable for long circulating therapeutic delivery vehicles.
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Metacrilatos/química , Micelas , Nanopartículas , Nanotecnologia , Fosforilcolina/análogos & derivados , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Fosforilcolina/química , Solventes/químicaRESUMO
Spectrally inactive, electrically insulating, and chemically inert are adjectives broadly used to describe phyllosilicate minerals like mica and chlorite. Here, the above is disproved by demonstrating aqueous suspensions of liquid exfoliated nanosheets from five bulk mica types and chlorite schist. Nanosheet quality is confirmed via transmission electron and X-ray photoelectron spectroscopies, as well as electron diffraction. Through Raman spectroscopy, a previously unreported size- and layer-dependent spectral fingerprint is observed. When analyzing the high-yield suspensions (≈1 mg mL-1 ) through UV-vis spectroscopy, all phyllosilicates present bandgap (Eg ) narrowing from ≈7 eV in the bulk to ≈4 eV for monolayers. Unusually, the bandgap is inversely proportional to the areal size (A) of the nanosheets, measured via atomic force microscopy. Due to an unrecorded quantum confinement effect, nanosheet electronic properties scale toward semiconducting behavior (bandgap ≈3 eV) as nanosheet area increases. Furthermore, modeling X-ray diffraction spectra shows that the root cause of the initial bandgap narrowing is lattice relaxation. Finally, with their broad range of isomorphically substituted ions, phyllosilicate nanosheets show remarkable catalytic properties for hydrogen production.
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Three-dimensional tissue scaffolds have utilised nanomaterials to great effect over the last decade. In particular, scaffold design has evolved to consider mechanical structure, morphology, chemistry, electrical properties, and of course biocompatibility - all vital to the performance of the scaffold and how successful they are in developing cell cultures. We have developed an entirely synthetic and tuneable three-dimensional scaffold of reduced graphene oxide (rGO) that shows good biocompatibility, and favourable mechanical properties as well as reasonable electrical conductivity. Importantly, the synthesis is scaleable and suitable for producing scaffolds of any desired geometry and size, and we observe a high level of biocompatibility and cell proliferation for multiple cell lines. In particular, one of the most devastating forms of malignant brain cancer, glioblastoma (GBM), grows especially well on our rGO scaffold in vitro, and without the addition of response-specific growth factors. We have observed that our scaffold elicits spontaneous formation of a high degree of intercellular connections across the GBM culture. This phenomenon is not well documented in vitro and nothing similar has been observed in synthetic scaffolds without the use of response-specific growth factors - which risk obscuring any potential phenotypic behaviour of the cells. The use of scaffolds like ours, which are not subject to the limitations of existing two-dimensional substrate technologies, provide an excellent system for further investigation into the mechanisms behind the rapid proliferation and success of cancers like GBM. These synthetic scaffolds can advance our understanding of these malignancies in the pursuit of improved theranostics against them.
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Materiais Biocompatíveis/química , Glioblastoma/metabolismo , Grafite/química , Alicerces Teciduais/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Condutividade Elétrica , Células Endoteliais da Veia Umbilical Humana , Humanos , PorosidadeRESUMO
Printed electronics based on liquid-exfoliated nanosheet networks are limited by inter-nanosheet junctions and thick films which hinder field-effect gating. Here, few-layer molybdenum disulfide nanosheets are assembled by Langmuir deposition into thin films, and size selection is shown to lead to a thousandfold conductivity enhancement with potential applicability to all nanosheet networks.
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Here, we develop a framework for assembly, understanding, and application of functional emulsions stabilized by few-layer pristine two-dimensional (2D) nanosheets. Liquid-exfoliated graphene and MoS2 are demonstrated to stabilize emulsions at ultralow nanosheet volume fractions, approaching the minimum loading achievable with 2D materials. These nanosheet-stabilized emulsions allow controlled droplet deposition free from the coffee ring effect to facilitate single-droplet devices from minute quantities of material or assembly into large-area films with high network conductivity. To broaden the range of compositions and subsequent applications, an understanding of emulsion stability and orientation in terms of surface energy of the three phases is developed. Importantly, this model facilitates determination of the surface energies of the nanosheets themselves and identifies strategies based on surface tension and pH to allow design of emulsion structures. Finally, this approach is used to prepare conductive silicone emulsion composites with a record-low loading level and excellent electromechanical sensitivity. The versatility of these nanosheet-stabilized emulsions illustrates their potential for low-loading composites, thin-film formation and surface energy determination, and the design of functional structures for a range of segregated network applications.
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Explosive percolation is an experimentally-elusive phenomenon where network connectivity coincides with onset of an additional modification of the system; materials with correlated localisation of percolating particles and emergent conductive paths can realise sharp transitions and high conductivities characteristic of the explosively-grown network. Nanocomposites present a structurally- and chemically-varied playground to realise explosive percolation in practically-applicable systems but this is yet to be exploited by design. Herein, we demonstrate composites of graphene oxide and synthetic polymer latex which form segregated networks, leading to low percolation threshold and localisation of conductive pathways. In situ reduction of the graphene oxide at temperatures of <150 °C drives chemical modification of the polymer matrix to produce species with phenolic groups, which are known crosslinking agents. This leads to conductivities exceeding those of dense-packed networks of reduced graphene oxide, illustrating the potential of explosive percolation by design to realise low-loading composites with dramatically-enhanced electrical transport properties.
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Although hot melt extrusion (HME) has been used in combination with fused deposition modelling (FDM) three-dimensional printing (3DP), suitable feedstock materials such as polymeric filaments with optimum properties are still limited. In this study, various release modifying excipients, namely, poly(vinyl alcohol) (PVA), Soluplus®, polyethylene glycol (PEG) 6000, Eudragit® RL PO/RS PO, hydroxypropyl methylcellulose (HPMC) K4M/E10M/K100M, Kollidon® vinyl acetate 64 (VA 64)/17PF/30, were used as a release modulating tool to control the drug release from 3D printed sustained release tablets. Ibuprofen (as the model drug) and ethyl cellulose (as the polymeric matrix), along with various release modifiers, were blended and extruded into filaments through a twin-screw extruder. Then these filaments were printed into cylindrical tablets through FDM 3DP technique and their surface morphology, thermal stability, solid-state, mechanical properties, dose accuracy and drug release behaviors were investigated. The solid-state analysis of 3D printed tablets exhibited the amorphous nature of the drug dispersed in the polymer matrices. Although all these prepared filaments could be successfully printed without failing during the FDM 3DP process, the mechanical characterization showed that the filament stiffness and brittleness could be adjusted significantly by changing the type of release modifiers. Moreover, in vitro drug release studies revealed that the drug release could simply be controlled over 24 h by only changing the type of release modifiers. All ibuprofen (IBP) loaded 3D printed tablets with ethyl cellulose (EC) matrix, especially with PEG as the release modifier, showed great potential in releasing IBP in a zero-order reaction. In conclusion, all the results illustrated that the HME/FDM approach and optimized formulation compositions can be an attractive option for the development of pharmaceutical tablets and implants where adjustable drug release patterns are necessary.
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Excipientes , Impressão Tridimensional , Liberação Controlada de Fármacos , Polímeros , Comprimidos , Tecnologia FarmacêuticaRESUMO
This study aims to prepare, optimize, and characterize magnetic-field-sensitive sugar-templated polydimethylsiloxane (PDMS) sponges for localized delivery of an anticancer drug, 5-fluorouracil (FLU). For this purpose, different concentrations of carbonyl iron (CI) and magnetite Fe3O4 nanopowders were embedded as magnetosensitive materials in PDMS resins for the fabrication of macroporous sponges via a sugar-template process. The process is environmentally friendly and simple. The fabricated interconnected macroporous magnetic particles loaded PDMS sponges possess flexible skeletons and good recyclability because of their recoverability after compression (deformation) without any breakdown. The prepared magnetic PDMS sponges were evaluated for their morphology (SEM and EDS), porosity (absorbency), elastic modulus, deformation under a magnetic field, thermostability, and in vitro cell studies. All physicochemical and magnetomechanical analysis confirmed that the optimized magnetic-field-sensitive PDMS sponge can provide an efficient method for delivering an on-demand dose of anticancer drug solutions at a specific location and timing with the aid of controlled magnetic fields.
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Antineoplásicos , Fluoruracila , Dimetilpolisiloxanos , Campos Magnéticos , PorosidadeRESUMO
To facilitate printable sensing solutions particles need to be suspended and stabilised in a liquid medium. Hansen parameters were used to identify that alcohol-water blends are ideal for stabilising colloidal copper hydroxide in dispersion. The suspended material can be further separated in various size fractions with a distinct cuboid geometry which was verified using atomic force microscopy. This facilitates the development of Raman spectroscopic metrics for determining particle sizes. This aspect ratio is related to the anisotropic crystal structure of the bulk crystallites. As the size of the nanocuboids decreases electrochemical sensitivity of the material increases due to an increase in specific surface area. Electrochemical glucose sensitivity was investigated using both cyclic voltammetry and chronoamperometry. The sensitivity is noted to saturate with film thickness. The electrochemical response of 253 mA M-1 cm-2 up to 0.1 mM and 120 mA cm-2 up to 0.6 mM allow for calibration of potential devices. These results indicate suitability for use as a glucose sensor and, due to the surfactant-free, low boiling point solvent approach used to exfoliate the nanocuboids, it is an ideal candidate for printable solutions. The ease of processing will also allow this material to be integrated in composite films for improved functionality in future devices.
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Cobre/química , Eletroquímica/métodos , Glucose/análise , Hidróxidos/química , Nanoestruturas/química , Eletroquímica/instrumentação , Eletrodos , Limite de Detecção , Solventes/químicaRESUMO
Starch-based excipients are commonly used in oral solid dosage forms. The effect of particle size and pregelatinisation level of starch-based excipients on their water absorption behaviour have been evaluated. The results showed that starch-based excipients have type ii isotherms, indicating that the principal mechanism of sorption is the formation of monolayer coverage and multilayer water molecules (10-80 RH %). It was found that the particle size of starch-based excipients did not have any influence on the rate of water sorption, whereas the level of pregelatinisation changed the kinetics of water sorption-desorption. Results showed that the higher the degree of pregelatinisation, the higher the rate of water absorption, which is irrespective of particle size. SEM images showed that a partially gelatinised starch had a firm granular structure with small pores and channels on the surface while a fully gelatinised starch had more irregular and spongy like surface with a degree of fractured particles.
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Excipientes/química , Amido/química , Vapor , Absorção Fisico-Química , Cinética , Modelos Químicos , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
Laser-deposited carbon aerogel is a low-density porous network of carbon clusters synthesized using a laser process. A one-step synthesis, involving deposition and annealing, results in the formation of a thin porous conductive film which can be applied as a chemiresistor. This material is sensitive to NO2 compared to ammonia and other volatile organic compounds and is able to detect ultra-low concentrations down to at least 10 parts-per-billion. The sensing mechanism, based on the solubility of NO2 in the water layer adsorbed on the aerogel, increases the usability of the sensor in practically relevant ambient environments. A heating step, achieved in tandem with a microheater, allows the recovery to the baseline, making it operable in real world environments. This, in combination with its low cost and scalable production, makes it promising for Internet-of-Things air quality monitoring.
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Nanohybrid materials based on nanoparticles of the intrinsically microporous polymer PIM-1 and graphene oxide (GO) are prepared from aqueous dispersions with a reprecipitation method, resulting in the surface of the GO sheets being decorated with nanoparticles of PIM-1. The significant blueshift in fluorescence signals for the GO/PIM-1 nanohybrids indicates modification of the optoelectronic properties of the PIM-1 in the presence of the GO due to their strong interactions. The stiffening in the Raman G peak of GO (by nearly 6 cm-1) further indicates p-doping of the GO in the presence of PIM. Kelvin probe force microscopy (KPFM) and electrochemical reduction measurements of the nanohybrids provide direct evidence for charge transfer between the PIM-1 nanoparticles and the GO nanosheets. These observations will be of importance for future applications of GO-PIM-1 nanohybrids as substrates and promoters in catalysis and sensing.
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A simple procedure of producing three-dimensional blisters of graphene through irradiation of the visible range laser by Raman spectrometer has been presented. Fabrication of different volumes of the blisters and their characterization were carried out with Raman spectroscopy by tuning the irradiation dose. The produced blisters showed a consistency in altitude and a remarkable change in functionality, adhesion force map and local contact potential difference as compared to untreated monolayer graphene and naturally occurred graphene nanobubbles. Nevertheless, bilayer graphene is unaffected in the applied laser doses. The laser irradiation led to lattice expansion of carbon atoms and introduced oxygenic functional groups with the structural disorder. The internal pressure of the gaseous molecules was evaluated by monitoring the shape of the graphene blisters and nanobubbles. High-resolution Raman mapping showed the impact of laser-affected area and the defect density (n d) is reported as a function of displacement. Our results reveal ease of applicability of the Raman laser for the imaging and texturing of graphene pointing toward the possibility of the desirable and cost-effective laser writing at the submicron scale by tuning photochemistry of graphene which is pivotal for numerous applications.
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Carbon nanofoam (CNF) is a highly porous, amorphous carbon nanomaterial that can be produced through the interaction of a high-fluence laser and a carbon-based target material. The morphology and electrical properties of CNF make it an ideal candidate for supercapacitor applications. In this paper, we prepare and characterize CNF supercapacitor electrodes through two different processes, namely, a direct process and a water-transfer process. We elucidate the influence of the production process on the microstructural properties of the CNF, as well as the final electrochemical performance. We show that a change in morphology due to capillary forces doubles the specific capacitance of the wet-transferred CNF electrodes.
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Inhalation of nanoparticles for pulmonary drug delivery offers the potential to harness nanomedicine formulations of emerging therapeutics, such as curcumin, for treatment of lung cancer. Biocompatible nanoparticles composed of poly(2-methacryloyloxyethyl phosphorylcholine)-b-poly(2-(diisopropylamino)ethyl methacrylate) (MPC-DPA) have been shown to be suitable nanocarriers for drugs, whilst N-trimethyl chitosan chloride (TMC) coating of nanoparticles has been reported to further enhance their cellular delivery efficacy; the combination of the two has not been previously investigated. Development of effective systems requires the predictable, controllable, and reproducible ability to prepare nanosystems possessing particle sizes, and drug loading capacities, appropriate for successful airway travel, lung tissue penetration, and tumor suppression. Although a number of MPC-DPA based nanosystems have been described, a complete understanding of parameters controlling nanoparticle formation, size, and morphology has not been reported; in particular the effects of differing solvents phases remains unclear. In this current study a matrix of 31 solvent combinations were examined to provide novel data pertaining to the formation of MPC-DPA nanoparticles, and in doing so afforded the selection of systems with particle sizes appropriate for pulmonary delivery applications to be loaded with curcumin, and coated with TMC. This paper presents the first report of novel data detailing the successful preparation, characterisation, and optimisation of MPC-DPA nanoparticles of circa 150-180nm diameter, with low polydispersity, and a curcumin loading range of circa 2.5-115µM, tunable by preparation parameters, with and without TMC coating, and thus considered suitable candidates for inhalation drug delivery applications.
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Portadores de Fármacos/química , Nanopartículas/química , Fosforilcolina/análogos & derivados , Ácidos Polimetacrílicos/química , Administração por Inalação , Quitosana/química , Curcumina/química , Microscopia Eletrônica de Transmissão e Varredura , Nanopartículas/ultraestrutura , Fosforilcolina/química , Solubilidade , Solventes/químicaRESUMO
OBJECTIVE: Uncontrolled rapid release of drugs can reduce their therapeutic efficacy and cause undesirable toxicity; however, controlled release from reservoir materials helps overcome this issue. The aims of this study were to determine the release profiles of ketoprofen and spironolactone from a pH-responsive self-assembling DPA-MPC-DPA triblock copolymer gel and elucidate underlying physiochemical properties. METHODS: Drug release profiles from DPA50 -MPC250 -DPA50 gel (pH 7.5), over 32 h (37 °C), were determined using UV-Vis spectroscopy. Nanoparticle size was measured by dynamic light scattering (DLS) and critical micelle concentration (CMC) by pyrene fluorescence. Polymer gel viscosity was examined via rheology, nanoparticle morphology investigated using scanning transmission electron microscopy (STEM) and the gel matrix observed using cryo-scanning electron microscopy (Cryo-SEM). KEY FINDINGS: DPA50 -MPC250 -DPA50 copolymer (15% w/v) formed a free-standing gel (pH 7.5) that controlled drug release relative to free drugs. The copolymer possessed a low CMC, nanoparticle size increased with copolymer concentration, and DLS data were consistent with STEM. The gel displayed thermostable viscosity at physiological temperatures, and the gel matrix was a nanostructured aggregation of smaller nanoparticles. CONCLUSIONS: The DPA50 -MPC250 -DPA50 copolymer gel could be used as a drug delivery system to provide the controlled drug release of ketoprofen and spironolactone.
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Sistemas de Liberação de Medicamentos/métodos , Cetoprofeno/química , Metacrilatos/química , Nanoestruturas/química , Fosforilcolina/análogos & derivados , Ácidos Polimetacrílicos/química , Espironolactona/química , Preparações de Ação Retardada/química , Preparações de Ação Retardada/metabolismo , Liberação Controlada de Fármacos , Géis , Cetoprofeno/metabolismo , Metacrilatos/metabolismo , Fosforilcolina/química , Fosforilcolina/metabolismo , Ácidos Polimetacrílicos/metabolismo , Espironolactona/metabolismoRESUMO
Proteus mirabilis forms extensive crystalline biofilms on indwelling urethral catheters that block urine flow and lead to serious clinical complications. The Bcr/CflA efflux system has previously been identified as important for development of P. mirabilis crystalline biofilms, highlighting the potential for efflux pump inhibitors (EPIs) to control catheter blockage. Here we evaluate the potential for drugs already used in human medicine (fluoxetine and thioridazine) to act as EPIs in P. mirabilis, and control crystalline biofilm formation. Both fluoxetine and thioridazine inhibited efflux in P. mirabilis, and molecular modelling predicted both drugs interact strongly with the biofilm-associated Bcr/CflA efflux system. Both EPIs were also found to significantly reduce the rate of P. mirabilis crystalline biofilm formation on catheters, and increase the time taken for catheters to block. Swimming and swarming motilies in P. mirabilis were also significantly reduced by both EPIs. The impact of these drugs on catheter biofilm formation by other uropathogens (Escherichia coli, Pseudomonas aeruginosa) was also explored, and thioridazine was shown to also inhibit biofilm formation in these species. Therefore, repurposing of existing drugs with EPI activity could be a promising approach to control catheter blockage, or biofilm formation on other medical devices.
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Infecções Relacionadas a Cateter/prevenção & controle , Fluoxetina/farmacologia , Infecções por Proteus/prevenção & controle , Proteus mirabilis/efeitos dos fármacos , Tioridazina/farmacologia , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Biofilmes/efeitos dos fármacos , Infecções Relacionadas a Cateter/microbiologia , Cateteres de Demora/efeitos adversos , Cateteres de Demora/microbiologia , Reposicionamento de Medicamentos , Fluoxetina/química , Humanos , Proteínas de Membrana Transportadoras/química , Proteínas de Membrana Transportadoras/metabolismo , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Infecções por Proteus/microbiologia , Proteus mirabilis/fisiologia , Tioridazina/química , Cateterismo Urinário/efeitos adversos , Cateterismo Urinário/instrumentação , Cateteres Urinários/efeitos adversos , Cateteres Urinários/microbiologiaRESUMO
Major challenges associated with nano-sized drug delivery systems include removal from systemic circulation by phagocytic cells and controlling appropriate drug release at target sites. 2-methacryloyloxyethyl phosphorylcholine (MPC) has been copolymerised in turn with two pH responsive comonomers (2-(diethylamino)ethyl methacrylate (DEA) and 2-(diisopropylamino)ethyl methacrylate (DPA), to develop novel biocompatible drug delivery vehicles. Micelles were prepared from a series of copolymers with varying block compositions and their colloidal stability and dimensions were assessed over a range of solution pH using photon correlation spectroscopy. The drug loading capacities of these micelles were evaluated using Orange OT dye as a model compound. The cytotoxicity of the micelles was assessed using an in vitro assay. The MPC-DEA diblock copolymers formed micelles at around pH 8 and longer DEA block lengths allowed higher drug loadings. However, these micelles were not stable at physiological pH. In contrast, MPC-DPA diblock copolymers formed micelles of circa 30 nm diameter at physiological pH. In vitro assays indicated that these MPC-DPA diblock copolymers had negligible cytotoxicities. Thus novel non-toxic biocompatible micelles of appropriate size and good colloidal stability with pH-modulated drug uptake and release can be readily produced using MPC-DPA diblock copolymers.
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Materiais Biocompatíveis/administração & dosagem , Sistemas de Liberação de Medicamentos , Metacrilatos/administração & dosagem , Nanoestruturas , Fosforilcolina/administração & dosagem , Concentração de Íons de Hidrogênio , Micelas , Tamanho da PartículaRESUMO
The comparison of copolymers containing sulfobetaine or phosphobetaine moieties for use as potential biocompatible coatings has been investigated. Two statistical copolymers were produced by a free radical polymerisation technique, one based on a sulfobetaine and the other on a phosphobetaine, both with a silyl group component to allow thermal crosslinking after coating. PMMA and glass discs were dip-coated with the polymers and their properties were compared to the uncoated controls. Bacterial adhesion to these coated materials was assessed using Staphylococcus epidermidis, Staphylococcus aureus and Pseudomonas aeruginosa. Human macrophages and granulocytes were used to assess the adhesion and activation of inflammatory cells whilst mouse 3T3 fibroblast cells were used to assess the propensity for the materials to support fibroblast cell adhesion. In all cases the polymer coatings reduced cell adhesion with respect to the base materials. The phosphobetaine-based copolymer coatings were shown to be markedly superior to the sulfobetaine-based copolymer coatings.
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Aderência Bacteriana/fisiologia , Betaína/química , Adesão Celular/fisiologia , Materiais Revestidos Biocompatíveis/química , Proteínas/química , Células 3T3 , Adsorção , Animais , Betaína/análogos & derivados , Células Cultivadas , Materiais Revestidos Biocompatíveis/síntese química , Reagentes de Ligações Cruzadas/química , Granulócitos/citologia , Granulócitos/fisiologia , Humanos , Macrófagos/citologia , Macrófagos/fisiologia , Teste de Materiais/métodos , CamundongosRESUMO
Proteus mirabilis is a common cause of catheter-associated urinary tract infections and frequently leads to blockage of catheters due to crystalline biofilm formation. Scanning electron microscopy (SEM) has proven to be a valuable tool in the study of these unusual biofilms, but entails laborious sample preparation that can introduce artefacts, undermining the investigation of biofilm development. In contrast, environmental scanning electron microscopy (ESEM) permits imaging of unprocessed, fully hydrated samples, which may provide much insight into the development of P. mirabilis biofilms. Here, we evaluate the utility of ESEM for the study of P. mirabilis crystalline biofilms in situ, on urinary catheters. In doing so, we compare this to commonly used conventional SEM approaches for sample preparation and imaging. Overall, ESEM provided excellent resolution of biofilms formed on urinary catheters and revealed structures not observed in standard SEM imaging or previously described in other studies of these biofilms. In addition, we show that energy-dispersive X-ray spectroscopy (EDS) may be employed in conjunction with ESEM to provide information regarding the elemental composition of crystalline structures and demonstrate the potential for ESEM in combination with EDS to constitute a useful tool in exploring the mechanisms underpinning crystalline biofilm formation.