Investigating the Molecular Mechanism of Protein-Polymer Binding with Direct Saturation Compensated Nuclear Magnetic Resonance.

Herein, we describe a new technique, direct saturation compensated transfer (DISCO) NMR, to characterize protein-macromolecule interactions. DISCO enables the direct observation of intermolecular interactions and is used to investigate mucoadhesion, a type of polymer-protein interaction that is widely implemented in drug delivery but remains poorly understood. In a model system of bovine submaxillary mucin and poly(acrylic acid), DISCO identifies selective backbone interactions that facilitate mucoadhesion through chain interpenetration. DISCO demonstrated distinct patterns of molecular selectivity between mucoadhesive polymers when applied to hydroxypropyl cellulose and carboxymethyl cellulose and that functionalizing adhesive polymers with strongly interacting moieties may be detrimental to the overall adhesive interaction. Additionally, DISCO was used to estimate polymer-protein dissociation constants using individual proton signals as reporters. Overall, DISCO can be used as a label-free screening tool to generate polymer-specific binding fingerprints to map and quantify interactions between macromolecules.

Low-Fouling Characteristics of Ultrathin Zwitterionic Cysteine SAMs.

Surface fouling remains an exigent issue for many biological implants. Unwanted solutes adsorb to reduce device efficiency and hasten degradation while increasing the risks of microbial colonization and adverse inflammatory response. To address unwanted fouling in modern implants in vivo, surface modification with antifouling polymers has become indispensable. Recently, zwitterionic self-assembled monolayers, which contain two or more charged functional groups but are electrostatically neutral and form highly hydrated surfaces, have been the focus of many antifouling coatings. Reports using various compositions of zwitterionic polymer brushes have demonstrated ultralow fouling in the ng/cm2 range. These coatings, however, are thick and can hinder the target application of biological devices. Here, we report an ultrathin (8.52 Å) antifouling self-assembled monolayer composed of cysteine that is amenable to facile fabrication. The antifouling characteristics of the zwitterionic surfaces were evaluated against bovine serum albumin, fibrinogen, and human blood in real time using quartz crystal microbalance and surface plasmon resonance imaging. Compared to untreated gold surfaces, the ultrathin cysteine coating reduced the adsorption of bovine serum albumin by 95% (43 ng/cm2 adsorbed) after 3 h and 90% reduction after 24 h. Similarly, the cysteine self-assembled monolayer reduced the adsorption of fibrinogen as well as human blood by >90%. The surfaces were further characterized using scanning electron microscopy: protein-enhanced adsorption and cellular adsorption in human blood was found on untreated surfaces but not on the cysteine SAM-protected surfaces. These findings suggest that surfaces can be functionalized with an ultrathin layer of cysteine to resist the adsorption of key proteins, with performance comparable to zwitterionic polymer brushes. As such, cysteine surface coatings are a promising methodology to improve the long-term utility of biological devices.

Discrimination of Proteins Using an Array of Surfactant-Stabilized Gold Nanoparticles.

Protein analysis is a fundamental aspect of biochemical research. Gold nanoparticles are an emerging platform for various biological applications given their high surface area, biocompatibility, and unique optical properties. The colorimetric properties of gold nanoparticles make them ideal for point-of-care diagnostics. Different aspects of gold nanoparticle-protein interactions have been investigated to predict the effect of protein adsorption on colloidal stability, but the role of surfactants is often overlooked, despite their potential to alter both protein and nanoparticle properties. Herein we present a method by which gold nanoparticles can be prepared in various surfactants and used for array-based quantification and identification of proteins. The exchange of surfactant not only changed the zeta potential of those gold nanoparticles but also drastically altered their aggregation response to five different proteins (bovine serum albumin, human serum albumin, immunoglobulin G, lysozyme, and hemoglobin) in a concentration-dependent manner. Finally, we demonstrate that varying surfactant concentration can be used to control assay sensitivity.

Optimization of Polydiacetylene-Coated Superparamagnetic Magnetite Biosensor for Colorimetric Detection of Biomarkers.

Biosensors for point-of-care testing of critical illnesses are urgently needed, especially in many areas of poor healthcare infrastructure. Polydiacetylene-based sensors are ideal because of their unique colorimetric properties where blue to red color shifts can be observed with the naked eye. In this work, a colorimetric biosensor capable of simple, rapid magnetic separation is optimized, using horse IgG as a model antibody, to obtain higher sensitivity. Composed of a unique combination of polydiacetylene and superparamagnetic iron oxide, the biosensor is fabricated at varying ratios of polydiacetylene to demonstrate optimization of color responsiveness. At increasing polydiacetylene ratios, improved color responsiveness and aqueous dispersion are observed, but the magnetic separation efficiency starts to suffer. The optimal color response is obtained at 90 wt% polydiacetylene. In addition, a 50 times improved lower detection limit of 0.01 mg/mL horse IgG is achieved, a relevant biomarker concentration for diagnosing sepsis. This platform provides a promising colorimetric biosensor for point-of-care use.

Nonfouling property of zwitterionic cysteine surface.

Applications of implantable bioelectronics for analytical and curative purposes are currently limited by their poor long-term biofunctionality in physiological media and nonspecific interactions with biomolecules. In an attempt to prolong in vivo functionality, recent advances in surface modifications have demonstrated that zwitterionic coatings can rival the performance of conventional poly(ethylene glycol) polymers in reducing nonspecific protein fouling. Herein, we report the fabrication of a very thin layer of nonfouling zwitterionic cysteine surface capable of protecting implantable bioelectronics from nonspecific adsorption of plasma proteins. This work is the first of its kind to fabricate, through solution chemistry, a cysteine surface exhibiting zwitterionic state as high as 88% and to demonstrate antibiofouling under the exposure of bovine serum albumin (BSA) and human serum. The fabricated surface utilized a minimal amount of gold substrate, approximately 10 nm, and an extremely thin antifouling layer at 1.14 nm verified by ellipsometry. X-ray photoelectron spectroscopy assessment of the nitrogen (N1s) and carbon (C1s) spectra conclude that 87.8% of the fabricated cysteine surface is zwitterionic, 2.5% is positively charged, and 9.6% is noncharged. Antibiofouling performance of the cysteine surface is quantitatively determined by bicinchoninic acid (BCA) protein assay as well as qualitatively confirmed using scanning electron spectroscopy. Cysteine surfaces demonstrated a BSA fouling of 3.9 ± 4.84% µg/cm(2), which is 93.6% and 98.5% lower than stainless steel and gold surfaces, respectively. Surface plasmon resonance imaging analysis returned similar results and suggest that a thinner cysteine coating will enhance performance. Scanning electron microscopy confirmed the results of BCA assay and suggested that the cysteine surface demonstrated a 69% reduction to serum fouling. The results reported in this paper demonstrate that it is possible to achieve a highly zwitterionic surface through solution chemistry on a macroscopic level that is capable of improving biocompatibility of long-term implantable bioelectronics.

Transfer-based nuclear magnetic resonance uncovers unique mechanisms for protein-polymer and protein-nanoparticle binding behavior.

Nanoparticle-based drug delivery systems have shown increasing popularity as a means to improve patient outcomes by improving the effectiveness of active pharmaceutical ingredients (APIs). Similarly, nanoparticles have shown success in targeting alternative routes of API administration, such as applying mucoadhesion or mucopenetration to mucosal drug delivery to enhance uptake. While there are many promising examples of mucoadhesive nanomedicines in literature, there are also many examples of contradictory mucoadhesive binding behavior, most prominently in cases using the same nanoparticle materials. We have uncovered mechanistic insights in polymer-protein binding systems using nOe transfer-based NMR and sought to leverage them to explore nanoparticle-protein interactions. We tested several polymer-coated nanoparticles and micellar polymer nanoparticles and evaluated their binding with mucin proteins. We uncovered that the composition and interaction intimacy of polymer moieties that promote mucin binding change when the polymers are incorporated onto nanoparticle surfaces compared to polymer in solution. This change from solution state to nanoparticle coating can enable switching of behavior of these materials from inert to binding, as we observed in polyvinyl pyrrolidone. We also found the nanoparticle core was influential in determining the binding fate of polymer materials, whereas the nanoparticle size did not possess a clear correlation in the ranges we tested (60-270 nm). These experiments demonstrate that identical polymers may switch their binding behavior to mucin as a function of conformational changes that are induced by incorporating the polymers onto the surface of nanoparticles. These NMR-derived insights could be further leveraged to optimize nanoparticle formulations and guide polymer-mediated mucoadhesion.

Transport and targeted binding of Pluronic-coated nanoparticles in unsaturated porous media.

The effectiveness of most in situ remedial technologies, including nanoremediation, lies on successful delivery of reagents to a subsurface target treatment zone. Targeted delivery of engineered nanoparticles (NPs) to treat petroleum hydrocarbons present in the unsaturated zone requires an understanding of their transport behaviour in these systems. A series of column experiments explored the effect of initial water saturation, flowrate, input dosage, and porous medium texture on the transport of iron oxide or cobalt ferrite NPs coated with an amphiphilic co-polymer, as well as their targeted attachment to a crude oil zone. As the initial water content increased with a concomitant reduction in air saturation, the degree of tailing present in the NP breakthrough curves (BTCs) reduced, and the mass of NPs recovered increased. Air saturation is positively correlated with the magnitude of air-water interfaces, which provide additional NP retention sites. At a lower injection flow rate, NP retention increased due to a longer residence time and comparatively high air saturation. NP transport behaviour was not sensitive to NP injection dose over the range tested. Increased retention and retardation of the NP BTC was observed in sediments with a higher clay and silt content. NPs coated with a lower concentration of a Pluronic block co-polymer to promote binding were preferentially retained within the crude oil zone. To simulate the asymmetrical NP breakthrough curves observed from the unsaturated systems required the use of a model that accounted for both mobile and immobile flow regions as well as NP attachment and detachment with nonlinear Langmuirian blocking. This model allowed examination of attachment and detachment rate coefficients which captured NP interaction with the porous medium and/or crude oil. It was found that the initial water saturation and flow rate did not have an appreciable impact on the NP attachment rate coefficient, while it increased by ~10× with increasing clay and silt content, and by ~100× in the presence of crude oil, indicating preferential NP attachment within the crude oil zone. As a result of the lower NP polymer concentration coating used to promote increased attachment to crude oil, higher retention was observed near the column inlet and was captured quantitatively by adding a depth-dependent straining term to the model. This retention behaviour represents a combination of irreversible attachment at the air-water interfaces and straining near the column inlet enhanced by the formation of NP aggregates. The detachment rate coefficient decreased with a lower initial water saturation and flowrate, but increased with higher clay and silt content. The findings from this study contribute to our understanding of the transport and binding behaviour of Pluronic-coated NPs in unsaturated conditions and, in particular, the role of initial water content, flowrate and porous medium texture. Demonstrated delivery of NPs to a target zone is an important step towards expanding the utility of NPs as treatment reagents.

Targeted delivery of cisplatin to prostate cancer cells by aptamer functionalized Pt(IV) prodrug-PLGA-PEG nanoparticles.

Cisplatin is used to treat a variety of tumors, but dose limiting toxicities or intrinsic and acquired resistance limit its application in many types of cancer including prostate. We report a unique strategy to deliver cisplatin to prostate cancer cells by constructing Pt(IV)-encapsulated prostate-specific membrane antigen (PSMA) targeted nanoparticles (NPs) of poly(D,L-lactic-co-glycolic acid) (PLGA)-poly(ethylene glycol) (PEG)-functionalized controlled release polymers. By using PLGA-b-PEG nanoparticles with PSMA targeting aptamers (Apt) on the surface as a vehicle for the platinum(IV) compound c,t,c-[Pt(NH(3))(2)(O(2)CCH(2)CH(2)CH(2)CH(2)CH(3))(2)Cl(2)] (1), a lethal dose of cisplatin was delivered specifically to prostate cancer cells. PSMA aptamer targeted delivery of Pt(IV) cargos to PSMA(+) LNCaP prostate cancer cells by endocytosis of the nanoparticle vehicles was demonstrated using fluorescence microscopy by colocalization of green fluorescent labeled cholesterol-encapsulated NPs and early endosome marker EEA-1. The choice of linear hexyl chains in 1 was the result of a systematic study to optimize encapsulation and controlled release from the polymer without compromising either feature. Release of cisplatin from the polymeric nanoparticles after reduction of 1 and formation of cisplatin 1,2-intrastrand d(GpG) cross-links on nuclear DNA was confirmed by using a monoclonal antibody for the adduct. A comparison between the cytotoxic activities of Pt(IV)-encapsulated PLGA-b-PEG NPs with the PSMA aptamer on the surface (Pt-NP-Apt), cisplatin, and the nontargeted Pt(IV)-encapsulated NPs (Pt-NP) against human prostate PSMA-overexpressing LNCaP and PSMA(-) PC3 cancer cells revealed significant differences. The effectiveness of PSMA targeted Pt-NP-Apt nanoparticles against the PSMA(+) LNCaP cells is approximately an order of magnitude greater than that of free cisplatin.

Precise engineering of targeted nanoparticles by using self-assembled biointegrated block copolymers.

There has been progressively heightened interest in the development of targeted nanoparticles (NPs) for differential delivery and controlled release of drugs. Despite nearly three decades of research, approaches to reproducibly formulate targeted NPs with the optimal biophysicochemical properties have remained elusive. A central challenge has been defining the optimal interplay of parameters that confer molecular targeting, immune evasion, and drug release to overcome the physiological barriers in vivo. Here, we report a strategy for narrowly changing the biophysicochemical properties of NPs in a reproducible manner, thereby enabling systematic screening of optimally formulated drug-encapsulated targeted NPs. NPs were formulated by the self-assembly of an amphiphilic triblock copolymer composed of end-to-end linkage of poly(lactic-co-glycolic-acid) (PLGA), polyethyleneglycol (PEG), and the A10 aptamer (Apt), which binds to the prostate-specific membrane antigen (PSMA) on the surface of prostate cancer (PCa) cells, enabling, respectively, controlled drug release, "stealth" properties for immune evasion, and cell-specific targeting. Fine-tuning of NP size and drug release kinetics was further accomplished by controlling the copolymer composition. By using distinct ratios of PLGA-b-PEG-b-Apt triblock copolymer with PLGA-b-PEG diblock copolymer lacking the A10 Apt, we developed a series of targeted NPs with increasing Apt densities that inversely affected the amount of PEG exposure on NP surface and identified the narrow range of Apt density when the NPs were maximally targeted and maximally stealth, resulting in most efficient PCa cell uptake in vitro and in vivo. This approach may contribute to further development of targeted NPs as highly selective and effective therapeutic modalities.

From prevention to diagnosis and treatment: Biomedical applications of metal nanoparticle-hydrogel composites.

Recent advances in biomaterials integrate metal nanoparticles with hydrogels to generate composite materials that exhibit new or improved properties. By precisely controlling the composition, arrangement and interactions of their constituents, these hybrid materials facilitate biomedical applications through myriad approaches. In this work we seek to highlight three popular frameworks for designing metal nanoparticle-hydrogel hybrid materials for biomedical applications. In the first approach, the properties of metal nanoparticles are incorporated into a hydrogel matrix such that the composite is selectively responsive to stimuli such as light and magnetic flux, enabling precisely activated therapeutics and self-healing biomaterials. The second approach mediates the dynamic reorganization of metal nanoparticles based on environment-directed changes in hydrogel structure, leading to chemosensing, microbial and viral detection, and drug-delivery capabilities. In the third approach, the hydrogel matrix spatially arranges metal nanoparticles to produce metamaterials or passively enhance nanoparticle properties to generate improved substrates for biomedical applications including tissue engineering and wound healing. This article reviews the construction, properties and biomedical applications of metal nanoparticle-hydrogel composites, with a focus on how they help to prevent, diagnose and treat diseases. Discussion includes how the composites lead to new or improved properties, how current biomedical research leverages these properties and the emerging directions in this growing field.

Factors affecting pluronic-coated iron oxide nanoparticle binding to petroleum hydrocarbon-impacted sediments.

Effective targeted delivery of nanoparticle agents may enhance the remediation of soils and site characterization efforts. Nanoparticles coated with Pluronic, an amphiphilic block co-polymer, demonstrated targeted binding behaviour toward light non-aqueous phase liquids such as heavy crude oil. Various factors including coating concentration, oil concentration, oil type, temperature, and pH were assessed to determine their effect on nanoparticle binding to heavy crude oil-impacted sandy aquifer material. Nanoparticle binding was increased by decreasing the coating concentration, increasing oil concentration, using heavier oil types, and increasing temperature, while pH over the range of 5-9 was found to have no effect. Nanoparticle transport and binding in columns packed with clean and oily porous media demonstrated the ability for efficient nanoparticle targeted binding. For the conditions explored, the attachment rate coefficient in columns packed with clean sand was 2.10 ± 0.66 × 10-4 s-1; however, for columns packed with oil-impacted sand a minimum attachment rate coefficient of 8.86 ± 0.43 × 10-4 s-1 was estimated. The higher attachment rate for the oil-impacted sand system indicates that nanoparticles may preferentially accumulate to oil-impacted zones present at heterogeneous impacted sites. Simulations were used to demonstrate this hypothesis using the set of parameters generated in this effort. This work contributes to our understanding of the application conditions that are required for efficient targeted binding of nanoparticles to crude-oil impacted porous media.

Targeted nanoparticle binding & detection in petroleum hydrocarbon impacted porous media.

Targeted nanoparticle binding has become a core feature of experimental pharmaceutical product design which enables more efficient payload delivery and enhances medical imaging by accumulating nanoparticles in specific tissues. Environmental remediation and geophysical monitoring encounter similar challenges which may be addressed in part by the adoption of targeted nanoparticle binding strategies. This study illustrates that engineered nanoparticles can bind to crude oil-impacted silica sand, a selective adsorption driven by active targeting based on an amphiphilic polymer coating. This coating strategy resulted in 2 mg/kg attachment to clean silica sand compared to 8 mg/kg attachment to oil-impacted silica sand. It was also shown that modifying the surface coating influenced the binding behaviour of the engineered nanoparticles - more hydrophobic polymers resulted in increased binding. Successful targeting of Pluronic-coated iron oxide nanoparticles to a crude oil and silica sand mixture was demonstrated through a combined quantitative Orbital Emission Spectroscopy mass analysis supported by Vibrating Scanning Magnetometer magnetometry, and a qualitative X-ray micro-computed tomography (CT) visualization approach. These non-destructive characterization techniques facilitated efficient analysis of nanoparticles in porous medium samples with minimal sample preparation, and in the case of X-Ray CT, illustrated how targeted nanoparticle binding may be used to produce 3-D images of contaminated porous media. This work demonstrated successful implementation of nanoparticle targeted binding toward viscous LNAPL such as crude oil in the presence of a porous medium, a step which opens the door to successful application of targeted delivery technology in environmental remediation and monitoring.

Tetanus toxin C fragment-conjugated nanoparticles for targeted drug delivery to neurons.

The use of nanoparticles for targeted drug delivery is often facilitated by specific conjugation of functional targeting molecules to the nanoparticle surface. We compared different biotin-binding proteins (avidin, streptavidin, or neutravidin) as crosslinkers to conjugate proteins to biodegradable nanoparticles prepared from poly(lactic-co-glycolic acid) (PLGA)-polyethylene glycol (PEG)-biotin polymers. Avidin gave the highest levels of overall protein conjugation, whereas neutravidin minimized protein non-specific binding to the polymer. The tetanus toxin C fragment (TTC), which is efficiently retrogradely transported in neurons and binds to neurons with high specificity and affinity, retained the ability to bind to neuroblastoma cells following amine group modifications. TTC was conjugated to nanoparticles using neutravidin, and the resulting nanoparticles were shown to selectively target neuroblastoma cells in vitro. TTC-conjugated nanoparticles have the potential to serve as drug delivery vehicles targeted to the central nervous system.

Formulation of functionalized PLGA-PEG nanoparticles for in vivo targeted drug delivery.

Nanoparticle (NP) size has been shown to significantly affect the biodistribution of targeted and non-targeted NPs in an organ specific manner. Herein we have developed NPs from carboxy-terminated poly(d,L-lactide-co-glycolide)-block-poly(ethylene glycol) (PLGA-b-PEG-COOH) polymer and studied the effects of altering the following formulation parameters on the size of NPs: (1) polymer concentration, (2) drug loading, (3) water miscibility of solvent, and (4) the ratio of water to solvent. We found that NP mean volumetric size correlates linearly with polymer concentration for NPs between 70 and 250 nm in diameter (linear coefficient=0.99 for NPs formulated with solvents studied). NPs with desirable size, drug loading, and polydispersity were conjugated to the A10 RNA aptamer (Apt) that binds to the prostate specific membrane antigen (PSMA), and NP and NP-Apt biodistribution was evaluated in a LNCaP (PSMA+) xenograft mouse model of prostate cancer. The surface functionalization of NPs with the A10 PSMA Apt significantly enhanced delivery of NPs to tumors vs. equivalent NPs lacking the A10 PSMA Apt (a 3.77-fold increase at 24h; NP-Apt 0.83%+/-0.21% vs. NP 0.22%+/-0.07% of injected dose per gram of tissue; mean+/-SD, n=4, p=0.002). The ability to control NP size together with targeted delivery may result in favorable biodistribution and development of clinically relevant targeted therapies.

Sustained release of bioactive therapeutic proteins from a biodegradable elastomeric device.

Effective localized delivery of a therapeutic protein requires a biodegradable device capable of delivering active protein at a sustained rate, and at a concentration within its therapeutic window. The objective of this study was to demonstrate that a biodegradable elastomeric device can be made in a cylindrical geometry, and still retain the ability to release a variety of therapeutic proteins at a nearly constant rate in nanomolar concentration with high bioactivity. The elastomers were prepared with cylindrical geometry by photo-cross-linking an acrylated star-poly(epsilon-caprolactone-co-d,l-lactide) macromer. Vascular endothelial growth factor (VEGF), interferon-gamma (IFN-gamma), and interleukin-2 (IL-2) were co-lyophilized with excipients, then entrapped within the elastomer matrix by photo-polymerization. Under identical formulation conditions, these proteins were released at the same, nearly constant rate for a significant part of the release profile (until 70%-80% release depending on formulation characteristics). Decreasing the molecular weight of the acrylated macromer increased the rate of protein release, but did not alter the zero order nature of the release kinetics. Cell based bioactivity assays showed only that 57% of the VEGF released was bioactive. By contrast, both IL-2 and IFN-gamma showed relatively high bioactivity and over 80% of the released proteins were bioactive. The elastomer formulation has potential as a regio-specific protein delivery device.

Maintenance of vascular endothelial growth factor and potentially other therapeutic proteins bioactivity during a photo-initiated free radical cross-linking reaction forming biodegradable elastomers.

Previously, we prepared a biodegradable elastomeric device that can release different therapeutic proteins at a nearly constant rate in nanomolar concentrations with high bioactivity. The elastomer device was fabricated using a photo-initiated free radical cross-linking reaction of acrylated star(epsilon-caprolactone-co-D,L-lactide) in organic solvent in the presence of solid protein particles. The objective of this study was to examine the effect of various parameters used for fabricating the photo-cross-linked elastomeric device on the stability of a therapeutic protein, vascular endothelial growth factor (VEGF), to determine which factor plays the dominant role in protecting VEGF. VEGF was lyophilized with or without bovine serum albumin (BSA) and then suspended in solid state in a macromer (acrylated star-poly(epsilon-caprolactone-co-D,L-lactide)) solution containing different concentrations of a free radical initiator, 2,2-dimethoxy-2-phenylacetophenone (DMPA). The protein suspension was then UV-irradiated at different intensities. UV irradiation with the generation of free radicals was detrimental to VEGF stability. BSA preserved the VEGF bioactivity during UV irradiation but provided little protection in the presence of the photo-initiator DMPA. The acrylated macromer acted as a free radical scavenger and effectively preserved VEGF and BSA stability during UV-initiated photo-polymerization. The detrimental effect of UV radiation with free radical generation on VEGF stability during device manufacture can be eliminated by choosing the proper bulking agents coupled with an efficient photo-polymerization reaction.

Controllable Microfluidic Production of Drug-Loaded PLGA Nanoparticles Using Partially Water-Miscible Mixed Solvent Microdroplets as a Precursor.

We present a versatile continuous microfluidic flow-focusing method for the production of Doxorubicin (DOX) or Tamoxifen (TAM)-loaded poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles (NPs). We use a partially water-miscible solvent mixture (dimethyl sulfoxide DMSO+ dichloromethane DCM) as precursor drug/polymer solution for NPs nucleation. We extrude this partially water-miscible solution into an aqueous medium and synthesized uniform PLGA NPs with higher drug loading ability and longer sustained-release ability than conventional microfluidic or batch preparation methods. The size of NPs could be precisely tuned by changing the flow rate ratios, polymer concentration, and volume ratio of DCM to DMSO (VDCM/VDMSO) in the precursor emulsion. We investigated the mechanism of the formation of NPs and the effect of VDCM/VDMSO on drug release kinetics. Our work suggests that this original, rapid, facile, efficient and low-cost method is a promising technology for high throughput NP fabrication. For the two tested drugs, one hydrophilic (Doxorubicin) the other one hydrophobic (Tamoxifen), encapsulation efficiency (EE) as high as 88% and mass loading content (LC) higher than 25% were achieved. This new process could be extended as an efficient and large scale NP production method to benefit to fields like controlled drug release and nanomedicine.

Interactions between bacterial surface and nanoparticles govern the performance of "chemical nose" biosensors.

Rapid and portable diagnosis of pathogenic bacteria can save lives lost from infectious diseases. Biosensors based on a "chemical nose" approach are attracting interest because they are versatile but the governing interactions between bacteria and the biosensors are poorly understood. Here, we use a "chemical nose" biosensor based on gold nanoparticles to explore the role of extracellular polymeric substances in bacteria-nanoparticle interactions. We employ simulations using Maxwell-Garnett theory to show how the type and extent of aggregation of nanoparticles influence their colorimetric response to bacteria. Using eight different species of Gram-positive and Gram-negative bacteria, we demonstrate that this "chemical nose" can detect and identify bacteria over two orders of magnitude of concentration (89% accuracy). Additionally, the "chemical nose" differentiates between binary and tertiary mixtures of the three most common hospital-isolated pathogens: Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa (100% accuracy). We demonstrate that the complex interactions between nanoparticles and bacterial surface determine the colorimetric response of gold nanoparticles and thus, govern the performance of "chemical nose" biosensors.

Sustained interferon-gamma delivery from a photocrosslinked biodegradable elastomer.

The application of protein therapeutics for long-term, localized delivery has been hindered by a lack of a delivery device that releases active protein at a concentration within their therapeutic window. A protein delivery system that uses an osmotic pressure delivery mechanism and a photocrosslinked biodegradable elastomer has been designed in an attempt to overcome this limitation. The elastomer is prepared through the UV initiated crosslinking of end terminal acrylated star-poly(epsilon-caprolactone-co-D,L-lactide). Interferon-gamma (IFN-gamma) was released from the optimum formulation at a constant rate of 23 ng/day over 21 days. A cell-based assay showed that over 83% of released IFN-gamma was bioactive. Furthermore, it was demonstrated that bovine serum albumin co-lyophilized with IFN-gamma was released at the same rate as IFN-gamma. This delivery formulation may be clinically useful for sustained, local protein drug delivery applications.

In vitro uptake and release of natamycin Dex-b-PLA nanoparticles from model contact lens materials.

PURPOSE: To evaluate the uptake and release of the antifungal agent natamycin encapsulated within poly(D,L-lactide)-dextran nanoparticles (Dex-b-PLA NPs) from model contact lens (CL) materials. METHODS: Six model CL materials (gel 1:poly(hydroxyethyl methacrylate, pHEMA); gel 2:85% pHEMA: 15% [Tris(trimethylsiloxy)silyl]-propyl methacrylate (TRIS); gel 3: 75% pHEMA: 25% TRIS; gel 4: 85% N,N dimethylacrylamide (DMAA): 15% TRIS; gel 5:75% DMAA: 25% TRIS; and gel 6: DMAA) were prepared using a photoinitiation procedure. The gels were incubated in: (1) natamycin dissolved in deionized (DI) water and (2) natamycin encapsulated within Dex-b-PLA NPs in dimethylsulfoxide/DI water. Natamycin release from these materials was monitored using UV-visible spectrophotometry at 304 nm over 7 d. RESULTS: Natamycin uptake by all model CL materials increased between 1 and 7 d (p < 0.001). The uptake of natamycin-NPs was higher than the uptake of the drug alone in DI water (p < 0.05). Drug release was higher in materials containing DMAA than pHEMA (p < 0.05). All gels loaded with natamycin-NPs also released more drug compared to gels soaked with natamycin in DI water (p < 0.001). After 1 h, CL materials loaded with natamycin alone released 28-82% of the total drug release. With the exception of gel 6, this burst released was reduced to 21-54% for CL materials loaded with natamycin-NPs. CONCLUSIONS: Model CL materials loaded with natamycin-Dex-b-PLA NPs were able to release natamycin for up to 12 h under infinite sink conditions. DMAA-TRIS materials may be more suitable for drug delivery of natamycin due to the higher drug release observed with these materials.