Vesicles, which include both liposomes and polymersomes (polymer vesicles), are being developed as therapeutic drug carriers. In this study, we present a fully scalable low pressure extrusion methodology for preparing vesicles. Vesicles were generated by continuous extrusion through a 200 nm pore diameter hollow fiber (HF) membrane. The first half of this study describes a method for generating empty polymersomes composed of different molecular weight amphiphilic poly(butadiene-b-ethylene oxide) (PBD-b-PEO) diblock copolymers on a large scale (50-100 mL) using a HF membrane. Monodisperse empty polymersomes were formed with particle diameters slightly less than 200 nm, which were close to the rated 200 nm pore size of the HF membrane. The second half of this study describes the successful encapsulation of hemoglobin (Hb) inside the aqueous core of polymersomes using the HF extrusion methodology. We demonstrate that polymersome encapsulated hemoglobin (PEH) particles formed by this technique had similar oxygen affinity, cooperativity coefficient, and methemoglobin (metHb) level compared to PEH particles formed by the 1 mL volume small scale manual extrusion method. Most notably, Hb encapsulation inside the polymer vesicles formed by the HF extrusion method increased 2-fold compared to the manual extrusion method. This work is important, since it will enable facile scale-up of homogeneous vesicle dispersions that are typically required for preclinical and clinical studies as well as industrial use.

A series of PEO/PLA copolymers, covering a wide range of compositions and segmental lengths, was synthesized, and their morphology was investigated by means of DSC and IR studies. For matrices comprising PEO chains with molecular weights below 3400, no soft-segment crystallinity was detected. When long hard segments were present, essentially monophasic, semicrystalline polymers were obtained, with PLA blocks melting around 130 degrees C. Polymers containing longer soft segments (PEO 6000) exhibited a two-phase matrix, with both components being able to crystallize. The relative degree of crystallinity of PEO and PLA blocks was also determined. The thermal history of the sample strongly affected the morphology of the matrix, especially when both blocks were long enough to crystallize. To further explore these polymers, solvent cast films were prepared and their morphology assessed. Casting from acetone, which is an excellent solvent for PLA, resulted in hard blocks exhibiting lower degrees of crystallinity, while methanol had a similar effect on PEO soft segments.

The aim of this work was to design injectable nanocarriers for drug delivery based on PCL-PEO amphiphilic block copolymers with linear ABA triblock and 4-armed (BA)(4) star-diblock architectures (A=PEO, B=PCL). The copolymers were obtained by coupling of a monofunctional -COOH end-capped PEO (M(n)=2.0kDa) with linear or 4-armed star-shaped PCL macromers bearing -OH terminal groups and were characterized by (1)H NMR spectroscopy and size exclusion chromatography. DSC and X-ray diffraction experiments showed that separate crystalline phases of PCL and PEO are present in bulk copolymers. Nanoparticles were produced by nanoprecipitation (NP) and by a new melting-sonication procedure (MS) without the use of toxic solvents, and characterized for size, polydispersity, zeta potential and core-shell structure. Nanoparticles were loaded with all-trans-retinoic acid (atRA) as a model drug and their release features assessed. Results demonstrate that both techniques allow the formation of PEO-coated nanoparticles with a hydrodynamic diameter that is larger for nanoparticles prepared by MS. atRA is released from nanoparticles at controlled rates depending on size, loading and, more important, preparation technique, being release rate faster for MS nanoparticles. Some biorelevant properties of the carrier such as complement activation were finally explored to predict their circulation time after intravenous injection. It is demonstrated that nanoparticles prepared by MS do not activate complement and are of great interest for future in vivo applications.

This research was conducted in order to fabricate stable polyethylene oxide (PEO)-based transmucosal systems of a Delta(9)-tetrahydrocannabinol (THC) prodrug, a hemisuccinate ester, using a hot-melt method. Since Delta(9)-tetrahydrocannabinol-hemisuccinate (THC-HS) was heat labile, a series of processing aids were evaluated in order to facilitate hot-melt production at lower temperatures, thereby reducing THC-HS degradation. The stability of THC-HS was influenced both by the processing conditions such as heating time and temperature, and the postprocessing storage conditions. The type of formulation additive also affected the extent of degradation. In the presence of polyethylene glycol (PEG)-400, the percentage of relative degradation of THC-HS to THC was 13.5% and 49.4% at 80 degrees C and 120 degrees C, respectively. In contrast, incorporation of vitamin E succinate (VES) reduced processing degradation to 2.1% and 9.2%, respectively, under the same conditions. Severe degradation of THC-HS was observed during storage, even under freezing conditions (-18 degrees C). A VES-Noveon AA-1 combination was observed to best stabilize the prodrug systems both during processing and postprocessing. Stabilization of THC-HS was achieved in these polyethylene oxide matrices at 4 degrees C, with almost 90% of theoretical drug remaining for up to 8 months. Investigation of the pH effect revealed that the pH of the microenvironment in these polymeric systems could be modulated to significantly improve the stability of THC-HS, degradation being the least in a relatively acidic medium.

In this work, PEO-α-CD pseudorotaxane hydrogels were prepared. The gels were loaded with proteins, BSA and lysozyme, representing proteins with different molecular weights. The kinetics of protein release was studied. Factors such as PEO concentration, protein concentration and exposed surface area of the gels were investigated to understand their effects on the release of the encapsulated cargo. Erosion of the gel surface appeared to be the dominant factor for release of the proteins. Fitting the data to various models supported our hypothesis that the mechanism of release was primarily erosion-driven as the data was best described by zero order, power law and Hopfenberg model. The linear relationship between the amount of mass loss and time establishes the erosion of the polymer gel matrix to be the key factor for drug release.

Small interfering RNA (siRNA) is a potential agent for the treatment of ocular surface diseases. Previous studies delivered siRNA by directly injecting siRNA into cornea or conjunctiva. In the present study we sought to explore an alternative approach to deliver siRNA into mouse cornea via eye drops that contains cy3-labeled siRNA (cy3-siRNA) and different cationic complexing agents and to evaluate the effects of siRNA targeting HSV-1 ICP4 gene (ICP4-siRNA) on mouse herpes simplex keratitis (HSK). Cy3-siRNA was mixed with Lipofectamine 2000, Entranster(TM)-in vivo, polyethyleneimine (PEI) or PEO-PPO-PEO polymers at different ratios. The efficacy of delivery was analyzed after topical application of the complexes to normal, EDTA treated, and epithelial scraped cornea of BALB/c mouse eyes. Compared to the other delivery agents and schedules, PEI at 0.75 mg/ml with 20 μM cy3-siRNA complex delivered eight times daily for two days was the most efficient as revealed by its production of the greatest fluorescence in cornea epithelial cells. In mouse HSK, the application of ICP4-siRNA+PEI eye drops reduced the damage to the corneal epithelia and decreased viral VP16 expression in the corneal tissue. These results proved the idea that siRNA can be formulated into eye drops with carriers for effective delivery into the cornea and that the formulated eye drops containing ICP4-siRNA can inhibit HSV-1 replication in the mouse corneas.

We present a neutron scattering analysis of the structure and dynamics of PEO polymer rings with a molecular weight 2.5 times higher than the entanglement mass. The melt structure was found to be more compact than a Gaussian model would suggest. With increasing time the center of mass (c.o.m.) diffusion undergoes a transition from sub-diffusive to diffusive behavior. The transition time agrees well with the decorrelation time predicted by a mode coupling approach. As a novel feature well pronounced non-Gaussian behavior of the c.o.m. diffusion was found that shows surprising analogies to the cage effect known from glassy systems. Finally, the longest wavelength Rouse modes are suppressed possibly as a consequence of an onset of lattice animal features as hypothesized in theoretical approaches.

The solubilization of a poorly water-soluble antiepileptic drug, carbamazepine (CBZ), in a series of micelle-forming PEO-PPO-PEO block copolymers with combinations of blocks having different molecular weight was studied. The drug solubility and micelle-water partition coefficient (P) were determined using UV-vis spectroscopy. Dynamic light scattering on copolymer solutions was used to measure size and polydispersity of nanoaggregates. Solubilization of carbamezapine increased with the rise in temperature and concentration of block copolymers, but no significant increase was observed with added salt (NaCl). The solubilization is also discussed from a thermodynamics viewpoint, by considering the standard free energy of solubilization (DeltaG degrees ).

Polystyrene (PS) latex particles of different sizes were adsorption coated with the polymeric surfactant Pluronic F108 (PEOPEOPEO tail mobility was investigated with electron paramagnetic resonance (EPR), and found to increase with an increase in polymer surface concentration. The comparatively slow motion of the PEO chains at low surface concentration indicated that not only the PPO block, but also the PEO blocks interacted hydrophobically with the PS surface. When the copolymer surface concentration was increased, the PEO tails were gradually being released, acquiring higher mobility as the surface became covered by the more strongly binding PPO blocks. Results obtained with F108 coated particles of different sizes showed that particle size had a significant effect on the fibrinogen uptake, with larger particles showing larger fibrinogen uptakes.

Within the framework of a proposed two-step mechanism for hydrate inhibition, the energy of binding of four inhibitor molecules (PEO, PVP, PVCap, and VIMA) to a hydrate surface is estimated with molecular dynamic simulations. One key feature of this proposed mechanism is that the binding of an inhibitor molecule to the surface of an ensuing hydrate crystal disrupts growth and therein crystallization. It is found through the molecular dynamic simulations that inhibitor molecules that experimentally exhibit better inhibition strength also have higher free energies of binding, an indirect confirmation of our proposed mechanism. Inhibitors increasing in effectiveness, PEO < PVP < PVCap < VIMA, have increasingly negative (exothermic) binding energies of -0.2 < -20.6 < -37.5 < -45.8 kcal/mol and binding free energies of increasing favorability (+0.4 approximately = +0.5 < -9.4 < -15.1 kcal/mol). Furthermore, the effect of an inhibitor molecule on the local liquid water structure under hydrate-forming conditions was examined and correlated to the experimental effectiveness of the inhibitors. Two molecular characteristics that lead to strongly binding inhibitors were found: (1) a charge distribution on the edge of the inhibitor that mimics the charge separation in the water molecules on the surface of the hydrate and (2) the congruence of the size of the inhibitor with respect to the available space at the hydrate-surface binding site. Equipped with this molecular-level understanding of the process of hydrate inhibition via low-dosage kinetic hydrate inhibitors we can design new, more effective inhibitor molecules.

OBJECTIVE

To correlate the molecular characteristics of the mtDNA deletions with clinical phenotypes.

METHODS

Southern analysis and polymerase chain reaction (PCR)/DNA sequencing were used to determine the size and location of deletions in 16 patients with mtDNA deletion syndrome. An additional 48 reported cases from the literature were also included in the statistical analysis.

RESULTS

The common 5-kb deletion is found in eight of nine patients with Kearns-Sayre syndrome (KSS), mitochondrial myopathies (MM), or progressive external ophthalmoplegia (PEO). The rare/novel deletions were found in six of seven patients with extra-neuromuscular multisystemic manifestations and infantile/early childhood onset.

CONCLUSIONS

Patients with mtDNA deletion syndrome who manifest non-neuromuscular multisystemic disorders at a very young age usually harbor mutant mtDNA with novel or rare deletions in every tissue analyzed. For this group of patients, it is possible to use the less invasive blood specimens instead of muscle biopsies for molecular diagnosis. Overwhelmingly, the common 5-kb deletion is mostly seen in the muscle specimens of patients with KSS and age of onset after the second decade of life.

Nonionic polyethylene oxide (PEO) and anionic polyacrylamide (PAM) flocculation of kaolinite dispersions has been investigated at pH 7.5 in the temperature range 20-60 degrees C. The surface chemistry (zeta potential), particle interactions (shear yield stress), and dewatering behavior were also examined. An increase in the magnitude of zeta potential of kaolinite particles, in the absence of flocculant and at a fixed PEO and PAM concentration, with increasing temperature was observed. The zeta potential behavior of the flocculated particles indicated a decrease in the adsorbed polymer layer thickness, while at the same time, however, the adsorbed polymer density showed a significant increase with increasing temperature. These results suggest that polymer adsorption was accompanied by temperature-influenced conformation changes. The hydrodynamic diameter and supernatant solution viscosity of both polymers decreased with increasing temperature, consistent with a change in polymer-solvent interactions and conformation, prior to adsorption. The analysis of the free energy (DeltaG(ads)) of adsorption showed a strong temperature dependence and the adsorption process to be more entropically than enthalpically driven. The polymer conformation change and increased negative charge at the kaolinite particle surface with increasing temperature resulted in decreased polymer bridging and flocculation performance. Consequently, the shear yield stress and the rate and the extent of dewatering (consolidation) of the pulp decreased significantly at higher temperatures (>40 degrees C). The temperature effect was more pronounced in the presence of PEO than PAM, with 40 and 20 degrees C indicated as the optima for enhanced performance of the latter and former flocculants, respectively. The results demonstrate that a temperature-induced conformation change, together with polymer structure type, plays an important role in flocculation and dewatering behavior of kaolinite dispersions.

Self-assembly of amphiphilic block co-polymers composed of poly(ethylene oxide) (PEO) as the hydrophilic block and poly(ether)s, poly(amino acid)s, poly(ester)s and polypropyleneoxide (PPO) as the hydrophobic block can lead to the formation of nanoscopic structures of different morphologies. These structures have been the subject of extensive research in the past decade as artificial mimics of lipoproteins and viral vectors for drug and gene delivery. The aim of this review is to provide an overview of the synthesis of commonly used amphiphilic block co-polymers. It will also briefly go over some pharmaceutical applications of amphiphilic block co-polymers as "nanodelivery systems" for small molecules and gene therapeutics.

We describe a series of surfactant polymers designed as surface-modifying agents for the suppression of bacterial adhesion on biomaterials. The surfactant polymers consist of a poly(vinyl amine) backbone with hydrophilic poly(ethylene oxide) (PEO) and hydrophobic hexanal (Hex) side chains (PVAm/PEO:Hex). Surface modification is accomplished by simple dip coating from aqueous solution, from which surfactant polymers undergo spontaneous surface-induced assembly on hydrophobic biomaterials. The stability of PVAm/PEO:Hex on pyrolytic graphite (HOPG) and polyethylene (PE) was demonstrated by the absence of detectable desorption under flow conditions of pure water over a 24-h period. PEO surfactant polymers with four different PEO:Hex ratios (1:1.4, 1:2.5, 1:4.6, and 1:10.7) and a dextran surfactant polymer were compared with respect to S. epidermidis adhesion under dynamic flow conditions. Suppression of S. epidermidis adhesion was achieved for all modified surfaces over the shear range 0-15 dyn/cm(2). The effectiveness depended on the surfactant polymer composition such that S. epidermidis adhesion to modified surfaces decreased significantly with increasing PEO packing density. Modified HOPG was more effective in reducing bacterial adhesion compared with the corresponding modification on PE, which we attribute to the presence of defects in surfactant polymer assembly on PE. Our results are discussed from the perspective of critical factors, such as optimal PEO packing density and hydration thickness, that contribute to the effectiveness of surfactant polymers to shield a biomaterial from adhesive bacterial interactions.

OBJECTIVE

The goal of the present study was to develop a poly (ethylene oxide) N10 (PEO N10) based melt-cast matrix system for efficient and prolonged delivery of hesperetin (HT), a promising bioflavonoid, to the posterior segment of the eye through the topical route.

METHODS

HT film was prepared by melt-cast method using PEO N10 and cut into 4mm×2mm segments, each weighing 8mg. This film was evaluated with respect to in vitro release rates and also transmembrane delivery across Spectra/Por® membrane (MWCO: 10,000 Daltons) and isolated rabbit corneas. Ocular tissue concentrations were also determined postapplication of the film in ex vivo and in vivo models.

RESULTS

HT release from the film was determined to be about 95.3% within 2h. In vitro transcorneal flux was observed to be 0.58±0.05μg/min/cm(2) across the isolated rabbit cornea. High levels of HT were detected in the retina-choroid (RC) and vitreous humor (VH) in the ex vivo model following topical application of the film. Significant levels of HT were observed in both anterior and posterior segment ocular tissues 1h post topical application of the 10 and 20%w/w HT films on the rabbit eye. Moreover, HT was detected in the VH and RC even after 6h following topical application of the film in vivo.

CONCLUSIONS

The results from this study suggest that the melt-cast films can serve as a viable platform for sustained topical delivery of bioflavonoids, and other therapeutic agents, into the back-of-the eye tissues.

The phase transfer of fluorescent CdSe based quantum dots (QDs) while retaining their properties and offering some advantages concerning the stability and functionalization characteristics is an important and intensively investigated field of research. Here we report how to tune and control the properties of CdSe/CdS/ZnS core-shell-shell QDs in water, using poly(isoprene-block-ethylene oxide) (PI-b-PEO) as a versatile system of amphiphilic diblock copolymers for the micellular encapsulation of nanoparticles (NPs). We show the synthesis of a novel PI-b-(PEO)2 miktoarm star polymer and how this different architecture besides the variation of the polymers' molecular weight gives us the opportunity to control the size of the built constructs in water between 24 and 53 nm. Because of this size control, an upper limit of the construct's diameter for the cellular uptake could be determined by a systemic study with human alveolar epithelial cells (A549) and murine macrophage leukemia cell (RAW-264.7). Furthermore, fluorescence quenching experiments with copper(II) and iron(III) ions show a strong influence of the used polymer on the shielding against these ions. This enables us to control the permeability of the polymer shell from very porous shells, which allow an almost complete cation exchange up to very dense shells. These even offer the possibility to perform copper(I) catalyzed click reactions while keeping the fluorescence of the QDs. All these results underline the huge variability and controllability of the PI-b-PEO diblock copolymer system for the encapsulation and functionalization of nanoparticles for biological applications. As a general trend, it can be stated that those coatings, which were most stable against quenchers, also showed the best resistivity with respect to unspecific cellular uptake.

We describe a two-generation family with combined clinical features of myoclonic epilepsy, progressive external ophthalmoplegia (PEO), proximal myopathy, pigmentary retinopathy, progressive deafness, basal ganglia calcification, and ragged-red fibers in a muscle biopsy specimen. One family member died unexpectedly at age 22 years. The molecular tests revealed an A-to-G transition at nucleotide position 3243 of the mitochondrial tRNA(Leu(UUR)) gene. No one in this family had stroke-like episodes. Although the propositus (a 28-year-old woman) had a significant number of white hairs, the percentage of mutant mtDNA in white-hair roots was not different from that in the colored-hair roots. Our findings suggest that the 3243 mutation can be associated with mixed clinical features of myoclonic epilepsy with ragged-red fibers (MERRF) and PEO and that a preferential increase in the levels of the mutant mtDNA is not related to graying of hair, and hence to the hypothesized production of premature aging of cells.

The evaluation of the severity of progressive external ophthalmoplegia (PEO) with ragged-red fibers in muscle, at the onset of the disease, when PEO is most often the only presenting symptom, is a difficult problem in neurological practice. In order to address that issue, we have performed a comparative analysis of the clinical, morphological and molecular characteristics of 43 patients affected with that form of ocular myopathy. Quantification of mitochondrial accumulation was performed with an image analysis application on muscle sections stained with succinate dehydrogenase histochemical reaction. The proportion of muscle fibres appearing as cytochrome c oxidase deficient was used as an index of the muscle-energy defect. Muscle mitochondrial DNA deletions were detected, localized and quantitated by Southern blot analysis. Point mutations were screened in five transfer RNA genes in the mtDNA (tRNA(Leucine (UUR)), tRNA(Lysine), tRNA(Glutamine), tRNA(Isoleucine) and tRNA(Formylmethionine)) by a denaturing gradient gel electrophoresis technique. This investigation confirmed the high frequency of mtDNA deletions or point mutations in PEO. At the onset of the disease, no clinical, morphological or molecular features could predict whether PEO would remain isolated or become part of a more severe multisystem disease. However, patients with mtDNA deletions were characterized by more severe ophthalmoplegia of earlier onset. Their muscle alterations were roughly parallel in severity to the proportion of deleted mtDNA molecules in muscle. Patients with a multitissular disease and mtDNA deletions were always sporadic cases and their clinical presentation was, most often, closely related to Kearns Sayre syndrome.

The significance of conserved cysteines in the human organic cation transporter 2 (hOCT2), namely the six cysteines in the long extracellular loop (loop cysteines) and C474 in transmembrane helix 11, was examined. Uptake of tetraethylammonium (TEA) and 1-methyl-4-phenypyridinium (MPP) into Chinese hamster ovary cells was stimulated >20-fold by hOCT2 expression. Both cell surface expression and transport activity were reduced considerably following mutation of individual loop cysteines (C51, C63, C89, C103, and C143), and the C89 and C103 mutants had reduced Michaelis constants (K(t)) for MPP. The loop cysteines were refractory to interaction with thiol-reactive biotinylation reagents, except after pretreatment of intact cells with dithiothreitol or following cell membrane solubilization. Reduction of disulfide bridge(s) did not affect transport, but labeling the resulting free thiols with maleimide-PEO(2)-biotin did. Mutation of C474 to an alanine or phenylalanine did not affect the K(t) value for MPP. In contrast, the K(t) value associated with TEA transport was reduced sevenfold in the C474A mutant, and the C474F mutant failed to transport TEA. This study shows that some but not all of the six extracellular loop cysteines exist within disulfide bridge(s). Each loop cysteine is important for plasma membrane targeting, and their mutation can influence substrate binding. The effect of C474 mutation on TEA transport suggests that it contributes to a TEA binding surface. Given that TEA and MPP are competitive inhibitors, the differential effects of C474 modification on TEA and MPP binding suggest that the binding surfaces for each are distinct, but overlapping in area.

We report on catechol-grafted poly(ethylene) glycol (PEG-g-catechol) for the preparation of nonfouling surfaces on versatile substrates including adhesion-resistant PTFE. PEG-g-catechol was prepared by the step-growth polymerization of PEO to which dopamine, a mussel-derived adhesive molecule, was conjugated. The immersion of substrates into an aqueous solution of PEG-g-catechol resulted in robust PEGylation on versatile surfaces of noble metals, oxides, and synthetic polymers. Surface PEGylation was unambiguously confirmed by various surface analytical tools such as ellipsometry, goniometry, infrared spectroscopy, and X-ray photoelectron spectroscopy. Contrary to existing PEG derivatives that are difficult-to-modify synthetic polymer surfaces, PEG-g-catechol can be considered to be a new class of PEGs for the facile surface PEGylation of various types of surfaces.

A novel bioeliminable amphiphilic poly(ethylene oxide)-b-poly(epsilon-caprolactone) (PEO-b-PCL) diblock copolymer end-capped by a mannose residue was synthesized by sequential controlled polymerization of ethylene oxide and epsilon-caprolactone, followed by the coupling of a reactive mannose derivative to the PEO chain end. The anionic polymerization of ethylene oxide was first initiated by potassium 2-dimethylaminoethanolate. The ring-opening polymerization of epsilon-caprolactone was then initiated by the omega-hydroxy end-group of PEO previously converted into an Al alkoxide. Finally, the saccharidic end-group was attached by quaternization of the tertiary amine alpha-end-group of the PEO-b-PCL with a brominated mannose derivative. The copolymer was fully characterized in terms of chemical composition and purity by high-resolution NMR spectroscopy and size exclusion chromatography. Furthermore, measurements with a pendant drop tensiometer showed that both the mannosylated copolymer and the non-mannosylated counterpart significantly decreased the dichloromethane/water interfacial tension. Moreover, these amphiphilic copolymers formed monodisperse spherical micelles in water with an average diameter of approximately 11 nm as measured by dynamic light scattering and cryo-transmission electron microscopy. The availability of mannose as a specific recognition site at the surface of the micelles was proved by isothermal titration microcalorimetry (ITC), using the BclA lectin (from Burkholderia cenocepacia), which interacts selectively with alpha-D-mannopyranoside derivatives. The thermodynamic parameters of the lectin/mannose interaction were extracted from the ITC data. These colloidal systems have great potential for drug targeting and vaccine delivery systems.

To reduce the incidence of postsurgical bacterial infection that may cause implantation failure at the implant-bone interface, surface treatment of titanium implants with antibiotic materials such as silver (Ag) has been proposed. The purpose of this work was to create TiO2 nanotubes using plasma electrolytic oxidation (PEO), followed by formation of an antibacterial Ag nanostructure coating on the TiO2 nanotube layer using a magnetron sputtering system. PEO was performed on commercially pure Ti sheets. The Ag nanostructure was added onto the resulting TiO2 nanotube using magnetron sputtering at varying deposition rates. Field emission scanning electron microscopy and transmission electron microscopy were used to characterize the surface, and Ag content on the TiO2 nanotube layer was analyzed by X-ray diffraction and X-ray photoelectron spectroscopy. Scanning probe microscopy for surface roughness and contact angle measurement were used to indirectly confirm enhanced TiO2 nanotube hydrophilicity. Antibacterial activity of Ag ions in solution was determined by inductively coupled plasma mass spectrometry and antibacterial testing against Staphylococcus aureus (S. aureus). In vitro, TiO2 nanotubes coated with sputtered Ag resulted in significantly reduced S. aureus. Cell viability assays showed no toxicity for the lowest sputtering time group in the osteoblastic cell line MC3T3-E1. These results suggest that a multinanostructured layer with a biocompatible TiO2 nanotube and antimicrobial Ag coating is a promising biomaterial that can be tailored with magnetron sputtering for optimal performance.

OBJECTIVE

To study the effects of hydrophobicity of the micelle-forming block copolymeric drug conjugate, methotrexate (MTX) esters of poly(ethylene oxide)-block-poly(2-hydroxyethyl-L-aspartamide) (MTX esters of PEO-b-PHEA), on the stability of micelles and on drug release.

METHODS

MTX esters of PEO-b-PHEA with three levels of MTX conjugation were synthesized. Size distribution of the micelles was measured by dynamic light scattering (DLS). The critical micelle concentration (CMC) was determined by a light scattering study. Size exclusion high performance liquid chromatography (SEC-HPLC) was used to study the equilibrium between unimers and micelles, and release of MTX at pH 7.4.

RESULTS

MTX esters of PEO-b-PHEA with MTX substitution of 7.4%, 22%, and 54% were prepared. The conjugates formed micelles based on DLS. The stability of the micelles correlated with the level of MTX conjugation. The conjugate with 54% MTX had a lower CMC (0.019 mg/mL) than the conjugates with 22% MTX (0.081 mg/mL) or 7.4% MTX (0.14 mg/mL). Micelle dissociation was significantly slower for the conjugate with 54% MTX than that with 22% and 7.4% MTX. Slower release of MTX from the micelles was also observed for the conjugate with the higher MTX attachment.

CONCLUSIONS

MTX esters of PEO-b-PHEA can be structurally modulated by varying the degree of MTX substitution, which in turn changes the hydrophobicity of the conjugate, thereby modifying micelle stability and controlling drug release.

The impact of mat porosity of polycaprolactone (PCL) electrospun fibers on the infiltration of neuron-like PC12 cells was evaluated using two different approaches. In the first method, bi-component aligned fiber mats were fabricated via the co-electrospinning of PCL with polyethylene oxide (PEO). Variation of the PEO flow rate, followed by selective removal of PEO from the PCL/PEO mesh, allowed for control of the porosity of the resulting scaffold. In the second method, aligned fiber mats were fabricated from various concentrations of PCL solutions to generate fibers with diameters between 0.13 ± 0.06 and 9.10 ± 4.1 μm. Of the approaches examined, the variation of PCL fiber diameter was found to be the better method for increasing the infiltration of PC12 cells, with the optimal infiltration into the ca. 1.5-mm-thick meshes observed for the mats with the largest fiber diameters, and hence largest pore sizes.