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Biomedical subjects

Anthony J Ryan

Publications and source records attributed to Anthony J Ryan.

14 recordsLinked to original sources
PubMed2007

Development of a 3D cell culture system for investigating cell interactions with electrospun fibers.

There are many variables to be considered in studying how cells interact with 3D scaffolds used in tissue engineering. In this study we investigated the influence of the fiber diameter and interfiber spaces of 3D electrospun fiber scaffolds on the behavior of human dermal fibroblasts. Fibers of two dissimilar model materials, polystyrene and poly-L-lactic acid, with a broad range of diameters were constructed in a specifically developed 3D cell culture system. When fibroblasts were introduced to freestanding fibers, and encouraged to "walk the plank," a minimum fiber diameter of 10 microm was observed for cell adhesion and migration, irrespective of fiber material chemistry. A distance between fibers of up to 200 microm was also observed to be the maximum gap that could be bridged by cell aggregates--a behavior not seen in conventional 2D culture. This approach has identified some basic micro-architectural parameters for electrospun scaffold design and some key differences in fibroblast growth in 3D. We suggest the findings will be of value for optimizing the integration of cells in these scaffolds for skin tissue engineering.

Biocompatible Materials↗
PubMed2006

Effect of substrate surface on dewetting behavior and chain orientation of semicrystalline block copolymer thin films.

Three symmetrical semicrystalline oxyethylene/oxybutylene block copolymers (EmBn) were spin-coated on different substrates including silicon, hydrophobically modified silicon, and mica. The effects of surface property on the dewetting behavior of EmBn thin films and the chain orientation of the crystalline block were investigated with atomic force microscopy and grazing incidence X-ray diffraction . The EmBn thin films on silicon exhibit an autophobic dewetting behavior, while ordinary dewetting occurs for the thin films on modified silicon. It was observed that the stems of the E crystals in the first half-polymer layer contacting the mica surface were parallel to the surface, in contrast to the perpendicular chain orientation of the other polymer layers and of the first half-polymer layer on silicon. This is attributed to the strong interaction between the E block and mica, verified by infrared spectra.

Journal Article↗
PubMed2006

Long-range structural order, moiré patterns, and iridescence in latex-stabilized foams.

We describe the facile production of highly stable foams stabilized solely by micrometer-sized, sterically stabilized polystyrene (PS) latex particles. Such foams can survive for more than one year in the wet state and remain intact after drying. In contrast, foams stabilized with either sodium dodecyl sulfate or poly(N-vinylpyrrolidone) were destroyed after removal of the aqueous phase. Scanning electron microscopy studies reveal hexagonally close-packed arrays of PS particles within the dried foam, which suggests high colloid stability for the PS particles prior to their adsorption at the air-water interface. Localized moiré patterns are observed by optical microscopy due to the formation of well-defined latex bilayers with exquisite long-range order. Moreover, the dried foams are highly iridescent in bright transmitted light, which may offer potential applications in security inks and coatings.

Journal Article↗
PubMed2006

Pathways of polymeric vesicle formation.

Polymeric vesicle formation is dictated by the mutual diffusion of water into the bulk block copolymer and vice versa. The hydration of three poly(ethylene oxide)-co-poly(butylene oxide) copolymers with different molecular weights has been monitored both macroscopically (confocal laser scanning microscopy) and microscopically (small-angle X-ray scattering). Both methods have revealed that the amphiphilic block copolymers swell in water following two qualitatively different growth regimes. Initially, water and copolymer diffuse into each other following a subdiffusional growth as the result of a molecular-level arrangement of the amphiphilic membranes that comprise the swollen copolymer. After a critical time, which is exponential in polymer molecular weight, the amphiphilic membranes reach their equilibrium morphology and as a consequence the growth starts to follow Fickian diffusion. The complex hydration kinetics dictate the phases formed at the interface between the amphiphilic copolymer and water. Upon hydration of simple amphiphiles, the amphiphilic film swells and the concentration gradient at the interface with water gradually drops to zero. This strongly affects the complex driving forces that control vesicle formation. Indeed, to form vesicles, an energy barrier has to be overcome, and therefore a constant concentration gradient is required. We show, by enhancing the hydration kinetics via an ac field, how the interface concentration gradient is kept constant and the magnitude of this gradient dictates the final size of the vesicles.

Journal Article↗
PubMed2006

Polymeric vesicle permeability: a facile chemical assay.

We present a simple method to characterize vesicles and determine, at the same time, the membrane permeability to specific molecules. The method is based on encapsulating highly hydrophilic 3,3',3' '-phosphinidynetris-benzenesulfonic acid (PH) into vesicles and subsequently monitoring its reaction with 5,5'-dithiobis-2-nitrobenzoic acid (DTNB). We tested the method by measuring the membrane permeability of vesicles formed from a series of poly(ethylene oxide)-co-polybutylene oxide (EB) copolymers and egg yolk phosphatidylcholine. We found that the experimental data are in good agreement with calculations based on Fick's first law. We therefore quantified the DTNB permeability across vesicle membranes, finding that polymeric EB membranes have a more selective permeability toward polar molecules compared to phospholipids membranes.

Dithionitrobenzoic Acid↗
PubMed2006

Reciprocating power generation in a chemically driven synthetic muscle.

A scalable synthetic muscle has been constructed that transducts nanoscale molecular shape changes into macroscopic motion. The working material, which deforms affinely in response to a pH stimulus, is a self-assembled block copolymer comprising nanoscopic hydrophobic domains in a weak polyacid matrix. A device has been assembled where the muscle does work on a cantilever and the force generated has been measured. When coupled to a chemical oscillator this provides a free running chemical motor that generates a peak power of 20 mW kg(-1) by the serial addition of 10 nm shape changes that scales over 5 orders of magnitude. It is the nanostructured nature of the gel that gives rise to the affine deformation and results in a robust working material for the construction of scalable muscle devices.

Biomechanical Phenomena↗
PubMed2005

Bilayers and interdigitation in block copolymer vesicles.

Amphiphilic block copolyethers assemble into membranes with thickness between 2.4 and 7.5 nm. The vesicular morphology has been confirmed by small-angle X-ray scattering combined with electron microscopy for diblock copolymers and triblock copolymers of both architectures. The scaling of the membrane thicknesses with the length of the hydrophobic block is in good agreement with the strong segregation theory for block copolymer melts, indicating a mixed and stretched conformation of the hydrophobic chain inside the vesicle membrane. This result is in contrast to previously published results where the hydrophobic membranes were observed to have bilayer geometry and polymer chains that are relatively unperturbed from their ideal Gaussian dimensions.

Biomimetic Materials↗
PubMed2005

The evolution of vesicles from bulk lamellar gels.

The remarkable ability of phospholipids to generate vesicles and lamellar morphologies has been mimicked by amphiphilic block copolymers and both classes of amphiphiles have been extensively studied in bulk and in dilute solution. The most common vesicle preparation method involves diffusion of water into a lamellar gel. The transformation proceeds from a collection of water-poor planar lamellae, to a water-rich interconnected layer-phase and then a collection of closed lamellae (vesicles) that form a close-packed gel. This is demonstrated by scattering and microscopy with the evolution of the gel structure being driven by increasing curvature. Vesicles have been observed to form when the copolymer-water system still has long-range order, with a discrete, clustered-vesicle structure leading to a turbid dispersion. Eventually, at very low concentration, the packed vesicles separate, generating very stable isotropic dispersions that are transparent. This phase sequence, involving a dispersion of close-packed vesicles, accounts for the formation of a narrow size distribution in these systems.

Evolution, Chemical↗
PubMed2003

Are metastable, precrystallisation, density-fluctuations a universal phenomena?

In-situ observations of crystallisation in minerals and organic polymers have been made by simultaneous, time-resolved small angle X-ray scattering (SAXS) and wide angle X-ray scattering (WAXS) techniques. In isotactic polypropylene slow quiescent crystallisation shows the onset of large scale ordering prior to crystal growth. Rapid crystallisations studied by melt extrusion indicate the development of well resolved oriented SAXS patterns associated with long range order before the development of crystalline peaks in the WAXS region. Block copolymers self-assemble into mesophases in polymer melts above a critical chain length (or above a critical temperature) and this self-assembly process is shown to be susceptible to an incipient crystallisation. Mesophase formation is observed at anomalously high temperatures in ethylene-oxide containing block copolymers below the normal melting point of the polyoxy ethylene chains. Formation of calcium carbonate from aqueous solutions of sodium carbonate and calcium nitrate is observed to be a two-stage process and precipitation proceeds by the production of an amorphous metastable phase. This phase grows until it is volume filling and leads to the formation of the two polymorphs Calcite and Vaterite. These three sets of results suggest pre-nucleation density fluctuations, leading to a metastable phase, play an integral role in all three classes of crystallisation. In due course, this phase undergoes transformation to "normal" crystals.

Journal Article↗
PubMedDate not supplied

Self-organization of skin cells in three-dimensional electrospun polystyrene scaffolds.

Much research in tissue engineering focuses on the synthesis of complex three-dimensional polymer scaffolds containing functional biomolecules to which cells are introduced. Typical scaffolds for skin tissue engineering are macroscopically porous with struts or fibers approximately 10 microm thick at a packing fraction of approximately 0.1. We made a polystyrene scaffold without cell signaling or spatial information by electrospinning and studied the growth of skin fibroblasts, keratinocytes, and endothelial cells, as single and cocultured populations in the presence and absence of fetal calf serum. In the absence of serum, keratinocytes, fibroblasts, and endothelial cells did not grow when cultured alone. However, when fibroblasts were cocultured with keratinocytes and endothelial cells, expansion of keratinocytes and endothelial cells occurred even in the absence of serum. Furthermore, cells displayed native spatial three-dimensional organization when cultured at an air-liquid interface, even when all three cell types were introduced at random to the scaffold. This study shows that coculture with fibroblasts enables keratinocytes and endothelial cells to proliferate without serum, but also to self-organize according to the native epidermal-dermal structure given the symmetry-breaking field of an air-liquid interface.

Biocompatible Materials↗
PubMedDate not supplied

Development of a closed bioreactor system for culture of tissue-engineered skin at an air-liquid interface.

A bioreactor has been developed for the production of tissue-engineered skin at an air-liquid interface for clinical and experimental use. In this closed system, scaffold and bioreactor sterilization, cell seeding, and medium perfusion were all performed with a peristaltic pump. Natural and synthetic dermal substitutes were seeded directly with skin cells without opening the bioreactor and fed either by continuous medium perfusion or by batch-feed. The system was validated by monoculture of human dermal fibroblasts and keratinocytes and the coculture of both cell types in acellular human dermis, Azowipes, electrospun polystyrene, and an electrospun composite of polystyrene and poly-DL-lactide fibers. A comparison was made of culture at an air-liquid interface versus submerged culture and of medium change by continuous perfusion versus batch-feed. Fibroblast and endothelial cells showed greater viability under submerged rather than air-liquid conditions whereas keratinocytes favored culture at an air-liquid interface as did cocultured keratinocytes and fibroblasts. Total cellular viability for reconstructed skin with keratinocytes and fibroblasts was greatest with continuous perfusion rather than batch-feed and with electrospun scaffolds compared with acellular human dermis. The bioreactor could also be easily configured to give replicate small areas for experimental use or one continuous area of construct for clinical use.

Air↗