Dialysate regeneration via urea photodecomposition with TiO2 nanowires at therapeutic rates.

BACKGROUND: The standard weekly treatment for end-stage renal disease patients is three 4-h-long hemodialysis sessions with each session c'onsuming over 120 L of clean dialysate, which prevents the development of portable or continuous ambulatory dialysis treatments. The regeneration of a small (~1 L) amount of dialysate would enable treatments that give conditions close to continuous hemostasis and improve patient quality of life through mobility. METHODS: Small-scale studies have shown that nanowires of TiO2 are highly efficient at photodecomposing urea into CO2 and N2 when using an applied bias and an air permeable cathode. To enable the demonstration of a dialysate regeneration system at therapeutically useful rates, a scalable microwave hydrothermal synthesis of single crystal TiO2 nanowires grown directly from conductive substrates was developed. These were incorporated into 1810 cm2 flow channel arrays. The regenerated dialysate samples were treated with activated carbon (2 min at 0.2 g/mL). RESULTS: The photodecomposition system achieved the therapeutic target of 14.2 g urea removal in 24 h. TiO2 electrode had a high urea removal photocurrent efficiency of 91%, with less than 1% of the decomposed urea generating NH4 + (1.04 µg/h/cm2 ), 3% generating NO3 - and 0.5% generating chlorine species. Activated carbon treatment could reduce total chlorine concentration from 0.15 to <0.02 mg/L. The regenerated dialysate showed significant cytotoxicity which could be removed by treatment with activated carbon. Additionally, a forward osmosis membrane with sufficient urea flux can cut off the mass transfer of the by-products back into the dialysate. CONCLUSION: Urea could be removed from spent dialysate at a therapeutic rate using a TiO2 based photooxidation unit, which can enable portable dialysis systems.

Targeted mRNA Therapy for Ornithine Transcarbamylase Deficiency.

We describe a novel, two-nanoparticle mRNA delivery system and show that it is highly effective as a means of intracellular enzyme replacement therapy (i-ERT) using a murine model of ornithine transcarbamylase deficiency (OTCD). Our Hybrid mRNA Technology delivery system (HMT) comprises an inert lipid nanoparticle that protects the mRNA from nucleases in the blood as it distributes to the liver and a polymer micelle that targets hepatocytes and triggers endosomal release of mRNA. This results in high-level synthesis of the desired protein specifically in the liver. HMT delivery of human OTC mRNA normalizes plasma ammonia and urinary orotic acid levels, and leads to a prolonged survival benefit in the murine OTCD model. HMT represents a unique, non-viral mRNA delivery method that allows multi-dose, systemic administration for treatment of single-gene inherited metabolic diseases.

Degradable, thermo-sensitive poly(N-isopropyl acrylamide)-based scaffolds with controlled porosity for tissue engineering applications.

We have developed a thermoresponsive poly(N-isopropyl acrylamide)-based scaffold with degradability and controlled porosity. Biodegradable poly(N-isopropyl acrylamide) hydrogels were synthesized by photocopolymerization of N-isopropylacrylamide with 2-methylene-1,3-dioxepane and polycaprolactone dimethacrylate. The hydrogels' phase transition temperature, swelling, and viscoelastic properties, as well as hydrolytic degradability at 25 and 37 °C, were explored. A sphere-templating technique was applied to fabricate hydrogel scaffolds with controllable pore size and a highly interconnected porous structure. The scaffold pore diameter change as a function of temperature was evaluated and, as expected, pores decreased in diameter when the temperature was raised to 37 °C. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test results suggested neither the scaffolds nor their degradation products were cytotoxic to NIH3T3 cells. Scaffolds with 55 ± 5 µm pore diameter were loaded with NIH3T3 cells and then were warmed to 37 °C entrapping cells in pores approximately 39 µm in diameter, a size range we have found to be optimal for angiogenesis and biointegration. Cells showed uniform infiltration and an elongated morphology after 7 days of culture. Due to the controlled monodisperse pore diameter, highly interconnected architecture, fully degradable chemistry and thermoresponsive properties, the polyNIPAM-based scaffolds developed here are attractive for applications in tissue engineering.

Radiopaque iodinated polymeric nanoparticles for X-ray imaging applications.

Iodinated radiopaque polymeric nanoparticles of sizes ranging between 30 and 350 nm were formed by emulsion polymerization of the monomer 2-methacryloyloxyethyl(2,3,5-triiodobenzoate) in the presence of sodium dodecyl sulfate as surfactant and potassium persulfate as initiator. The influence of various polymerization parameters, e.g., monomer, initiator and surfactant concentrations on the molecular weight, polymerization yield, size and size distribution of the particles was elucidated. Characterization of these iodinated nanoparticles was accomplished by routine methods such as FTIR, 1H NMR, TEM, TGA, DSC, GPC and light scattering. These polymeric nanoparticles are composed of ca. 58% by weight iodine, and are therefore expected to possess significant radiopaque nature. In vitro radiopacity of the iodinated nanoparticles of 30.6+/-5.0 nm diameter, dispersed in water and in the dry state, was demonstrated with a CT scanner. In vivo CT-imaging performed in a dog model by intravenous administration of the uniform 30.6+/-5.0 nm diameter radiopaque nanoparticles dispersed in saline demonstrated significant enhanced visibility of lymph nodes, liver, kidney and spleen. These results indicate that these nanoparticles may be useful as new efficient contrast agents for X-ray imaging applications.

Synthesis and characterization of radiopaque magnetic core-shell nanoparticles for X-ray imaging applications.

Radiopaque magnetic gamma-Fe(2)O(3)/poly(2-methacryloyloxyethyl(2,3,5-triiodobenzoate)) core-shell nanoparticles of narrow size distribution were prepared by emulsion polymerization of the iodinated monomer 2-methacryloyloxyethyl(2,3,5-triiodobenzoate) in the presence of maghemite (gamma-Fe(2)O(3) nanoparticles coated with a dextran shell are commonly used as contrast agents for magnetic resonance imaging (MRI). The present nanoparticles have similar core-shell structure substituting the dextran for the iodo polymer. These core-shell nanoparticles may therefore be useful as imaging contrast agents to detect various pathogenic zones and to observe different disease states in both modes: X-ray and MRI.

Synthesis and characterization of uniform radiopaque polystyrene microspheres for X-ray imaging by a single-step swelling process.

Uniform radiopaque polystyrene microspheres of approximately 2.3 +/- 0.2 microm were prepared by a single-step swelling of 2.3 +/- 0.2 microm polystyrene template microspheres, dispersed in an aqueous solution with methylene chloride emulsion droplets containing 2,3,5-triiodobenzoylethyl ester. After completing the swelling process, the methylene chloride was evaporated in order to lock the 2,3,5-triiodobenzoylethyl ester in the polystyrene microspheres. The influence of the weight ratio [2,3,5-triiodobenzoylethyl ester]/[polystyrene] on the % of entrapped 2,3,5-triiodobenzoylethyl ester was elucidated. Characterization of the radiopaque polystyrene microspheres was accomplished by light microscope, FTIR, TGA, SEM, XPS, and elemental analysis. The radiopacity of the microspheres was demonstrated by an imaging technique based on X-ray absorption usually used in hospitals. This novel method of encapsulation of 2,3,5-triiodobenzoylethyl ester within polystyrene microspheres by a single-step swelling process may be used as a model for encapsulation of various hydrophobic radiopaque drugs within uniform polystyrene template particles of various diameters for different X-ray imaging needs, e.g., blood pool, body organs, embolization, dental composition, implants, protheses, and nanocomposites.

Drug encapsulated aerosolized microspheres as a biodegradable, intelligent glioma therapy.

The grim prognosis for patients diagnosed with malignant gliomas necessitates the development of new therapeutic strategies for localized and sustained drug delivery to combat tumor drug resistance and regrowth. Here we introduce drug encapsulated aerosolized microspheres as a biodegradable, intelligent glioma therapy (DREAM BIG therapy). DREAM BIG therapy is envisioned to deliver three chemotherapeutics, temporally staged over one year, via a bioadhesive, biodegradable spray directly to the brain surgical site after tumor excision. In this proof-of-principle article exploring key components of the DREAM BIG therapy prototype, rhodamine B (RB) encapsulated poly(lactic-co-glycolic acid) and immunoglobulin G (IgG) encapsulated poly(lactic acid) microspheres were formulated and characterized. The encapsulation efficiency of RB and IgG and the release kinetics of the model drugs from the microspheres were elucidated in addition to the release kinetics of RB from poly(lactic-co-glycolic acid) microspheres formulated in a degradable poly(N-isopropylacrylamide) solution. The successful aerosolized application onto brain tissue ex-vivo demonstrated the conformal adhesion of the RB encapsulated poly(lactic-co-glycolic acid) microspheres to the convoluted brain surface mediated by the thermoresponsive carrier, poly(N-isopropylacrylamide). These preliminary results suggest the potential of the DREAM BIG therapy for future use with multiple chemotherapeutics and microsphere types to combat gliomas at a localized site.

Drug encapsulated polymeric microspheres for intracranial tumor therapy: A review of the literature.

Despite intensive surgical excision, radiation therapy, and chemotherapy, the current life expectancy for patients diagnosed with glioblastoma multiforme is only 12 to 15months. One of the approaches being explored to increase chemotherapeutic efficacy is to locally deliver chemotherapeutics encapsulated within degradable, polymeric microspheres. This review describes the techniques used to formulate drug encapsulated microspheres targeted for intracranial tumor therapy and how microsphere characteristics such as drug loading and encapsulation efficiency can be tuned based on formulation parameters. Further, the results of in vitro studies are discussed, detailing the varied drug release profiles obtained and validation of drug efficacy. Finally, in vivo results are summarized, highlighting the study design and the effectiveness of the drug encapsulated microspheres applied intracranially.

Precision-Porous PolyHEMA-Based Scaffold as an Antibiotic-Releasing Insert for a Scleral Bandage.

Combat-related penetrating ocular injuries have become a common form of battlefield injury in modern warfare and can lead to potentially devastating visual impairments. Prompt stabilization of the wounded globe via prevention of infection and fibrosis enhances the probability of a successful outcome after professional medical treatment. In this study, a norfloxacin-releasing poly(hydroxyethyl methacrylate)-based insert was designed and fabricated as a part of scleral bandage to prevent development of infection and scar formation. First, a sphere-templating technique was applied, during which 2-hydroxyethyl methacrylate monomer was photocopolymerized with acrylic acid and/or 4-fluorostyrene at different molar ratios to generate poly(hydroxyethyl methacrylate)-based porous scaffolds of various compositions with interconnected, monodisperse, 38 µm diameter pores. The scaffolds were then loaded with norfloxacin via swelling in drug-saturated solutions of various solvents, such as water, acetone, chloroform and ethanol, and the effect of the scaffold composition and the swelling solvent on norfloxacin uptake was explored. An in vitro drug release study was then conducted to explore the release kinetics of norfloxacin from the drug-loaded scaffolds, with the aim to find the optimal scaffold composition to provide release of norfloxacin over a 1 week period. The antibacterial potential of the optimal composition norfloxacin-loaded scaffold to inhibit the growth of the relevant clinical pathogens Staphylococcus epidermidis and Pseudomonas aeruginosa during a 1 week period was evaluated in vitro using a continuous culture flow cell system and a soft agar overlay plate assay.

Synthesis and fabrication of a degradable poly(N-isopropyl acrylamide) scaffold for tissue engineering applications.

Biodegradable poly(N-isopropyl acrylamide) (polyNIPAM) hydrogels with controlled molecular weight of the parent polymer and its degradation products were synthesized by atom transfer radical polymerization in the presence of a polycaprolactone-based di-chlorinated macroinitiator and polycaprolactone dimethacrylate. The phase transition temperature, swelling, hydrolytic degradability, and mechanical properties at 25 and 37°C were explored. A cytocompatibility study showed good NIH3T3 cell response over 5 days culture on the surface of the hydrogels, demonstrated by a consistent increase in cell proliferation detected by an Alamar Blue assay. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] results suggested that the hydrogels and their degradation products in the concentration range of 1-25 mg/mL were not cytotoxic to NIH3T3 cells. A sphere-templating technique was utilized to fabricate biodegradable polyNIPAM scaffolds with monodisperse, pore size. Scaffolds with pore diameter of 48 ± 6 µm were loaded with A-10 smooth muscle cells and then warmed to 37°C entrapping cells in pores approximately 40 µm in diameter, a size we have found to be optimal for angiogenesis and biointegration. Due to their degradable nature, tunable molecular weight, highly interconnected morphology, thermally controlled monodisperse pore size, and temperature-induced volume expansion-contraction, the polyNIPAM-based scaffolds developed in this work will be valuable in tissue engineering.

Integrated bi-layered scaffold for osteochondral tissue engineering.

Osteochondral tissue engineering poses the challenge of combining both cartilage and bone tissue engineering fundamentals. In this study, a sphere-templating technique was applied to fabricate an integrated bi-layered scaffold based on degradable poly(hydroxyethyl methacrylate) hydrogel. One layer of the integrated scaffold was designed with a single defined, monodispersed pore size of 38 µm and pore surfaces coated with hydroxyapatite particles to promote regrowth of subchondral bone while the second layer had 200 µm pores with surfaces decorated with hyaluronan for articular cartilage regeneration. Mechanical properties of the construct as well as cyto-compatibility of the scaffold and its degradation products were elucidated. To examine the potential of the biphasic scaffold for regeneration of osteochondral tissue the designated cartilage and bone layers of the integrated bi-layered scaffold were seeded with chondrocytes differentiated from human mesenchymal stem cells and primary human mesenchymal stem cells, respectively. Both types of cells were co-cultured within the scaffold in standard medium without soluble growth/differentiation factors over four weeks. The ability of the integrated bi-layered scaffold to support simultaneous matrix deposition and adequate cell growth of two distinct cell lineages in each layer during four weeks of co-culture in vitro in the absence of soluble growth factors was demonstrated.

Synthesis and characterization of recombinant factor VIIa-conjugated magnetic iron oxide nanoparticles for hemophilia treatment.

Maghemite (gamma-Fe(2)O(3)) nanoparticles of 15.0 +/- 2.1 nm in diameter were prepared by nucleation, followed by controlled growth of magnetic iron oxide thin films onto gelatin nuclei. Functionalization of these magnetic nanoparticles with activated double bonds was accomplished by interacting divinyl sulfone with the gelatin coating of the gamma-Fe(2)O(3) nanoparticles. The activated double bonds were then used for covalent binding, via Michael addition reaction, of recombinant factor VIIa and human serum albumin to the surface of these nanoparticles. Recombinant factor VIIa was also physically bound to the magnetic nanoparticles by interacting this factor with the human serum albumin conjugated gamma-Fe(2)O(3) nanoparticles. The influence of factor VIIa concentration on the immobilization yield has been elucidated. Leakage of the bound factor VIIa into PBS containing 4% albumin was insignificant. The coagulant activity of the physically adsorbed recombinant factor VIIa was similar to that of the free one and was significantly better than that of the covalently bound. The blood half-life of free factor VIIa is short, about 2-3 h, because of digestion by proteolytic enzymes and inhibitory effects. Stabilization of factor VIIa against trypsin (a model proteolytic enzyme) and chloromethyl ketone-type inhibitor was accomplished by conjugation of the factor to the gamma-Fe(2)O(3) nanoparticles. This stabilization may extend the blood half-life of factor VIIa. Therefore, IV injection of factor VIIa conjugated gamma-Fe(2)O(3) nanoparticles instead of free factor may avoid the frequent dosing and reduce the cost of hemophilia treatment.

Synthesis and characterization of new micrometer-sized radiopaque polymeric particles of narrow size distribution by a single-step swelling of uniform polystyrene template microspheres for X-ray imaging applications.

Radiopaque micron-sized non-cross-linked polystyrene/poly(2-methacryloyloxyethyl(2,3,5-triiodobenzoate)) particles of narrow size distribution were prepared by a single-step swelling of uniform polystyrene template microspheres with emulsion droplets of methylene chloride containing the initiator benzoyl peroxide and the iodinated monomer 2-methacryloyloxyethyl(2,3,5-triiodobenzoate), followed by the polymerization of the monomer within the swollen template particles at 73 degrees C. Radiopaque micron-sized uniform cross-linked polystyrene/poly(2-methacryloyloxyethyl(2,3,5-triiodobenzoate)-divinylbenzene) composite particles were prepared similarly with emulsion droplets of methylene chloride containing divinylbenzene, in addition to the initiator and the iodinated monomer. Radiopaque micron-sized uniform cross-linked poly(2-methacryloyloxyethyl(2,3,5-triiodobenzoate)-divinylbenzene) particles were formed by dissolving the template polystyrene polymer belonging to the former cross-linked composite particles. Characterization of these novel radiopaque polymeric particles was performed by methods such as FTIR, TGA, DSC, SEM, XPS, elemental analysis, and light microscopy. The influence of the weight ratio [2-methacryloyloxyethyl(2,3,5-triiodobenzoate)]/[polystyrene] and [2-methacryloyloxyethyl(2,3,5-triiodobenzoate)]/[divinylbenzene] on the bulk and surface properties of the non-cross-linked and cross-linked particles, respectively was elucidated. The radiopacity of these iodinated particles was demonstrated by an imaging technique based on X-ray absorption usually used in hospitals. These novel radiopaque particles may be used for different X-ray imaging needs, e.g., blood pool, body organs, embolization, dental composition, implants, protheses, and nanocomposites.