Nanospheres

An approach for enzyme therapeutics is elaborated with cell-implanted nanoreactors that are based on multiple enzymes encapsulated in hollow silica nanospheres (HSNs). The synthesis of HSNs is carried out by silica sol-gel templating of water-in-oil microemulsions so that polyethyleneimine (PEI) modified enzymes in aqueous phase are encapsulated inside the HSNs. PEI-grafted superoxide dismutase (PEI-SOD) and catalase (PEI-CAT) encapsulated in HSNs are prepared with quantitative control of the enzyme loadings. Excellent activities of superoxide dismutation by PEI-SOD@HSN are found and transformation of H2 O2 to water by PEI-CAT@HSN. When PEI-SOD and PEI-CAT are co-encapsulated, cascade transformation of superoxide through hydrogen peroxide to water was facile. Substantial fractions of HSNs exhibit endosome escape to cytosol after their delivery to cells. The production of downstream reactive oxygen species (ROS) and COX-2/p-p38 expression show that co-encapsulated SOD/CAT inside the ...

We present a novel approach for analysis of low-conductivity and insulating materials with conventional pulsed-voltage atom probe tomography (APT), by incorporating an ultrathin metallic coating on focused ion beam prepared needle-shaped specimens. Finite element electrostatic simulations of coated atom probe specimens were performed, which suggest remarkable improvement in uniform voltage distribution and subsequent field evaporation of the insulated samples with a metallic coating of approximately 10nm thickness. Using design of experiment technique, an experimental investigation was performed to study physical vapor deposition coating of needle specimens with end tip radii less than 100nm. The final geometries of the coated APT specimens were characterized with high-resolution scanning electron microscopy and transmission electron microscopy, and an empirical model was proposed to determine the optimal coating thickness for a given specimen size. The optimal coating strategy was ...

In this report, we demonstrate glucose-derived carbon nanospheres 1 to be an emerging class of intracellular carriers. The surfaces of these spheres are highly functionalized and do not need any further modification. Besides, the intrinsic fluorescence property of carbon nanospheres helps in tracking their cellular localization without any additional fluorescent tags. The spheres are found to target the nucleus of the mammalian cells, causing no toxicity. Interestingly, the in vivo experiments show that these nanospheres have an important ability to cross the blood-brain barrier and localize in the brain besides getting localized in the liver and the spleen. There is also evidence to show that they are continuously being removed from these tissues over time. Furthermore, these nanospheres were used as a carrier for the membrane-impermeable molecule CTPB (N-(4-chloro-3-trifluoromethylphenyl)-2-ethoxybenzamide), the only known small-molecule activator of histone acetyltransferase (HAT) p300. 2 Biochemical analyses such as Western blotting, immunohistochemistry, and gene expression analysis show the induction of the hyperacetylation of histone acetyltransferase (HAT) p300 (autoacetylation) as well as histones both in vitro and in vivo and the activation of HAT-dependent transcription upon CTPB delivery. These results establish an alternative path for the activation of gene expression mediated by the induction of HAT activity instead of histone deacetylase (HDAC) inhibition.

The purpose of this work was to explore the relation between protein-protein interactions (PPIs) and solution viscosity at high protein concentration using three monoclonal antibodies (mAbs), two of the IgG 4 subclass and one of the IgG 1 subclass. A range of methods was used to quantify the PPI either at low concentration (interaction parameter (k D ) obtained from dynamic light scattering, DLS) or at high concentration (solution storage modulus (G 0 ) from ultrasonic shear rheology). We also developed a novel method for the determination of PPI using the apparent radius of the protein at either low or high protein concentration determined using DLS. The PPI measurements were correlated with solution viscosity (measured by DLS using polystyrene nanospheres and ultrasonic shear rheology) as a function of pH (4-9) and ionic strength (10, 50 and 150 mM). Our measurements showed that the highest solution viscosity was observed under conditions with the most negative k D , the highest apparent radius and the lowest net charge. An increase in ionic strength resulted in a change in the nature of the PPI at low pH from repulsive to attractive. In the neutral to alkaline pH region the mAbs behaved differently with respect to increase in ionic strength. Two mAbs (A and B) showed little or no effect of increasing ionic strength, whereas mAb-C showed a remarkable decrease in attractive PPI and viscosity.

Anatase TiO2 displaying dual morphologies were synthesized with a simple chemical route via a single step. A strong correlation between the dye sensitized solar cell (DSSC) device performance and the obtained dual morphologies are highlighted using relevant evidences from UV-vis and diffuse reflectance spectroscopy (DRS), Raman analysis, field emission scanning electron microscope, high resolution transmission electron microscope, current-voltage characteristics and impedance analysis. Structural investigation revealed that the interested medium in this work employed for the synthesis of TiO2 produced different types of dual morphologies such as nanospheres with nanoparticles (NSNP), microsphere decorated with nanoparticles (μSNP) and nanoparticles with stone like features (NPS). UV-vis and DRS results indicated the ability of the materials to assist with diffused reflectance and optical absorption of the screen printed photoanodes. In addition, an investigation of Raman vibrational characteristics was carried out in detail using synchronous and asynchronous 2D correlation analysis. The high intense Eg anatase mode of Raman spectrum was simulated using a phonon confinement model and its normalised spectra are compared with an experimental data. The resultant performances in the DSSCs were not indicative by the material properties; so, the charge transfer resistance (Rct) and recombination properties of the photoanode were investigated. These results showed that the Nyquist plot of NSNP possesses the smallest diameter, which reveals that the device based on NSNP offers the lowest Rct value that accounts for the higher conversion efficiencies when compared with the DSSC device based on photoanodes μSNP and NSP. It can be concluded that the improved power conversion efficiency shown by photoanode fabricated using NSNP has the dual role of optimum surface area for dye loading and light scattering centres. These properties, when optimized along with the offered pathway for electrolyte diffusion to the dye degeneration with less Rct, could lead to better photoelectric conversion efficiency.

In the recent past, the targeted drug delivery has gained more attention for various advantages. Amongst the plethora of Avenues explored for targeted drug delivery. Nanoparticles are particulate dispersions or solid particles with a size in the range of 10-1000nm. The drug is dissolved, entrapped, encapsulated or attached to a nanoparticle matrix. Depending upon the method of preparation, nanoparticles, nanospheres or nanocapsules can be obtained. The major goals in designing nanoparticles as a delivery system are to control particle size, surface properties and release of pharmacologically active agents in order to achieve the site-specific action of the drug at the therapeutically optimal rate and dose regimen. Present review reveals the methods of preparation, characterization and application of several nanoparticulate drug delivery systems.

Since the last decade, nanohole arrays have emerged from an interesting optical phenomenon to the development of applications in photophysical studies, photovoltaics and as a sensing template for chemical and biological analyses. Numerous methodologies have been designed to manufacture nanohole arrays, including the use of focus ion beam milling, soft-imprint lithography, colloidal lithography and, more recently, modified nanosphere lithography (NSL). With NSL or colloidal lithography, the experimental conditions control the density of the nanosphere mask and, thus, the aspect of the nanohole arrays. Low surface coverage of the nanosphere mask produces disordered nanoholes. Ordered nanohole arrays are obtained with a densely packed nanosphere mask in combination with electrochemical deposition of the metal, glancing angle deposition (GLAD) or etching of the nanospheres prior to metal deposition. A review of these methodologies is presented here with an emphasis on the optical properties of nanoholes interesting in analytical chemistry. In particular, applications of these novel plasmonic materials will be demonstrated as substrates for a localized surface plasmon resonance (LSPR), Surface Plasmon Resonance (SPR), surface enhanced Raman spectroscopy (SERS), and in electrochemistry with nano-patterned electrodes.

The purpose of this research was to develop polylactic-co-glycolic acid (PLGA) nanospheres surface modified with chitosan (CS). Mitoxantrone-(MTO-) loaded PLGA nanospheres were prepared by a solvent evaporation technique. The PLGA nanospheres surface was modified with CS by two strategies (adsorption and covalent binding). PLGA nanospheres of 248.4 ± 21.0 nm in diameter characterized by the laser light scattering technique, scanning electron microscopy (SEM) are spherical and its drug encapsulation efficiency is 84.1 ± 3.4%. Zeta potential of unmodified nanospheres was measured to be negative −21.21 ± 2.13 mV. The positive zeta potential of modified nanospheres reveals the presence of CS on the surface of the modified nanospheres. Modified nanospheres were characterized for surface chemistry by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FT-IR). FT-IR spectra exhibited peaks at 3420 cm −1 and 1570 cm −1 , XPS spectra shows the N 1s (atomic orbital 1s of nitrogen) region of the surface of the nanospheres, corresponding to the primary amide of CS. In vitro drug release demonstrated that CS-modified nanospheres have many advantages such as prolonged drug release property and decreased the burst release over the unmodified nanospheres, and the modified nanospheres by covalent binding method could achieve the release kinetics of a relatively constant release. These data demonstrate high potential of CS-modified PLGA nanospheres for the anticancer drug carrier.

For the first time, uniform molecularly imprinted polymer (MIP) nanoparticles were prepared using dicamba as a template. The MIP nanoparticles were successfully synthesized by precipitation polymerization using methacrylic acid (MAA) as functional monomer, trimethylolpropane trimethacrylate (TRIM) as cross-linker and acetonitrile as porogen. The produced polymers were characterized by differential scanning calorimetry (DSC) and their morphology was precisely examined by scanning electron microscopy (SEM). The MIP nanospheres were obtained with the average diameter of 234 nm. Batch-wise guest binding experiments were carried out to determine the removal efficiency of the produced MIP nanoparticles towards the template molecule in aqueous solutions. The MIP showed outstanding affinity toward dicamba in aqueous solution with maximum removal efficiency of 87.5% at 300 mg.L-1 of dicamba solution. The MIP exhibited higher adsorption efficiency compared with the corresponding non-imprinted polymer (NIP) as well as outstanding selectivity towards dicamba relative to the template analog in an aqueous solution. Moreover, effects of pH on removal efficiency and selectivity of MIP were evaluated in detail.

A one-step process for the production of nanoparticles presenting advanced magnetic properties can be achieved using vapor condensation. In this article, we report on the fabrication of Fe particles covered by a uniform MgO epitaxial shell. MgO has a lower surface energy than Fe, which results in a core-shell crystal formation. The particles satisfy a few of technical requirements for the practical use in real clinics, such as a high biocompatibility in living cells in-vitro, an injection through blood vessels without any clothing problems in murine model, a high absorption rate for magnetic hyperthermia at small particle concentration, and the potential to be used as contrast agent in the field of diagnostic magnetic imaging. They are also able to be used in drug delivery and magnetic-activated cell sorting.

Potential mechanisms for formation of highly organized biomineralized structures include oriented crystal growth on templates, the aggregation of nanocrystals by oriented attachment, and the assembly of inorganic nanoparticles mediated by organic molecules into aggregated structures. In the present study, the potential role of amorphous calcium phosphate (ACP) in facilitating the assembly of hydroxyapatite (HAP) nanoparticles into highly ordered structures was evaluated. The physical characteristics of HAP nanoparticles prepared by three different methods were analyzed after extended exposure to additives in solution. Higher order HAP architecture was detected only when the starting particles were aggregates of nanospheres with HAP cores and ACP shells. Enamel-like HAP architecture was produced when the biologic additive was 10 mM glycine or 1.25 µM amelogenin. Large platelike crystals of the type present in bone were induced when the additive was 10 mM glutamic acid. Surface ACP initially links the HAP nanoparticles in a way that allows parallel orientation of the HAP nanoparticles and then is incorporated into HAP by phase transformation to produce a more highly ordered architecture with features that are characteristic for HAP in biologic structures. These studies provide evidence for a new mechanism for assembly of biominerals in which ACP functions by linking HAP nanocrystals while they assume parallel orientations and then is incorporated by phase transformation into HAP molecules that rigidly link HAP nanocrystals in larger fused crystallites. Biologic molecules present during this process of biomineral assembly specifically regulate the assembly kinetics and determine the structural characteristics of the final HAP architecture.

6 b S Supporting Information 23 24 ' INTRODUCTION 25 Surface-enhanced Raman scattering (SERS) is a promising 26 technique for the simultaneous detection of multiplex target 27 molecules. In contrast to the broad and relatively featureless spectra 28 of fluorescence labels, SERS labels produce signals with very 29 narrow bandwidth and fingerprint-like information for analytes, 30 which give them a high multiplexing capacity. Other advantages 31 of SERS include lack of photobleaching and self-quenching, as well 32 as the use of a single excitation source for multiple species. 1À3 In 33 particular, SERS nano tags are playing an increasingly important 34 role in bioassays and biomedical diagnostics, including for detec-35 tion of DNA and RNA, 4À6 proteins, 3,7À9 viral pathogens, 10 and 36 cellular imaging. 11À16 Specific nanostructures that can provide a 37 high level of electromagnetic field enhancement are a prerequisite 38 in order to obtain stable spectroscopic signatures and sensitive 39 detection. 17 With the exception of the commonly used Ag and Au 40 nanoparticles, various nanostructures have been used to prepare 41 SERS tags, such as Au nanorods, 18 nanoshells, 19À21 hollow gold 42 nanosphere, 22À24 AuÀAg nanocages, 25 and Au nanostars. 26 43 Most of the SERS nano tags developed to date have taken 44 advantage of the high sensitivity offered by SERS and the specificity 45 offered by surface probing molecules. However, a common draw-46 back of these approaches is that the tags are not isolated from the 47 environment and thus are easily influenced by surrounding con-48 ditions. For instance, signals from cell components, in addition to 49 SERS signals generated by reporter molecules, have been ob-50 served from unprotected SERS tags used for pH sensing in living 51 cells. 27 Therefore, it is necessary to prepare SERS nanoparticle 52 tags with protective shells, which can prevent desorption of 53 Raman reporters and adsorption of external species. To this end, 54 several approaches have been developed to prepare polymer-or 55 silica-encapsulated nanoparticle SERS tags. Polymer-encapsu-56 lated nanoparticle SERS tags have been prepared by using thiol-57 modified polyethylene glycol as a stabilizer and protective 58 shell. 9,14 Likewise, metal nanoparticles developed from polystyr-59 ene-block-poly(acrylic acid) (PS 154 -b-PAA 60 ), an amphiphilic di-60 block copolymer, through self-assembly have also been reported. 28 61 However, due to its optical transparency, easy surface functio-62 nalization, and biocompatibility, silica has been considered a 63 good coating material for the synthesis of SERS nano tags. 64 Mulvaney and co-workers 29 were the first to prepare "glass-65 ABSTRACT: The use of silica shells offers many advantages in 8 surface-enhanced Raman scattering (SERS)-based biological 9 sensing applications due to their optical transparency, remark-10

Since it was invented two decades ago, Nanosphere Lithography (NSL) has been widely studied as a low cost and flexible technique to fabricate nanostructures. Based on the registered patents and some selected papers, this review will discuss recent developments of different NSL strategies for the fabrication of ordered nanostructure arrays. The mechanism of self-assembly process and the techniques for preparing the self-assembled nanosphere template are first briefly introduced. The nanosphere templates are used either as shadow masks or as moulds for pattern transfer. Much more work now combines NSL with other lithographic techniques and material growth methods to form novel nanostructures of complex shape or various materials. Hence, this review finally gives a discussion on some future directions in NSL study.

We present a luminescence oxygen sensor incorporated in a wireless intraocular microrobot for minimallyinvasive diagnosis. This microrobot can be accurately controlled in the intraocular cavity by applying magnetic fields. The microrobot consists of a magnetic body susceptible to magnetic fields and a sensor coating. This coating embodies Pt(II) octaethylporphine (PtOEP) dyes as the luminescence material and polystyrene as a supporting matrix, and it can be wirelessly excited and read out by optical means. The sensor works based on quenching of luminescence in the presence of oxygen. The excitation and emission spectrum, response time, and oxygen sensitivity of the sensor were characterized using a spectrometer. A custom device was designed and built to use this sensor for intraocular measurements with the microrobot. Due to the intrinsic nature of luminescence lifetimes, a frequencydomain lifetime measurement approach was employed. An alternative sensor implementation using poly(styrene-co-maleic anhydride) (PS-MA) and PtOEP was successfully demonstrated with nanospheres to increase sensor performance.

The objective of this work was to develop a modified release system for the local anesthetic lidocaine (LDC), using poly(g-caprolactone) (PCL) nanospheres (NSs), to improve the pharmacological properties of the drug when administered by the infiltration route. In vitro experiments were used to characterize the system and investigate the release mechanism. The NSs presented a polydispersion index of 0.072, an average diameter of 449.6 nm, a zeta potential of −20.1 mV, and an association efficiency of 93.3%. The release profiles showed that the release of associated LDC was slower than that of the free drug. Atomic force microscopy analyses showed that the spherical structure of the particles was preserved as a function of time, as well as after the release experiments. Cytotoxicity and pharmacological tests confirmed that association with the NSs reduced the toxicity of LDC, and prolonged its anesthetic action. This new formulation could potentially be used in applications requiring gradual anesthetic release, especially dental procedures.

We report on the optical characterization of three-dimensional opal-like photonic crystals made by self-organized nanospheres of poly[styrene-(co-2-hydroxyethyl methacrylate)] having a face centred cubic (fcc) structure oriented along the [111] direction. A detailed optical characterization of the samples is presented using angle resolved reflection spectroscopy in specular geometry. The investigated energies are between a/λ=0.5 and a/λ=1.5 (where a is the lattice parameter and λ is the light wavelength), a region in which both first and second-order Bragg diffraction are expected. Some interesting features as branching of the Bragg peak dispersion and high energy reflection peaks are revealed. We compare the experimental data with theoretical calculations using both Bragg diffraction and band structure approach. A comparison with recent results reported in the literature is also presented.

Strangles is an infectious disease caused by Streptococcus equi subspecies equi that affects the upper respiratory tract of the Equidae. The control of this disease seems to be dependent on its earlier detection and prevention, but prolonged animal protection without development of strong and severe side effects has not yet been achieved. Convalescent horses exhibit a protective immune response, mainly against SeM (58 kDa), an antiphagocytic and opsonogenic S. equi M-like protein, known as the major protective antigen against strangles. Purified recombinant SeM and S. equi protein extract-entrapped poly(lactic acid) (PLA) nanospheres were developed and their adjuvant potential was studied via the intramuscular route. The effect including molecules with adjuvant properties such as spermine, oleic acid, alginate and glycol-chitosan was also evaluated. Spherical nanometric particles <500 nm containing the protein antigen were prepared by the solvent evaporation method and protein structure was not affected throughout preparation. The humoral immune response induced by nanospheres was markedly higher than that elicited by soluble antigens, isolated or co-admixed with CpG. The IgG and IgG subtypes, along with cytokine titres, indicated that nanospheres composed by glycolchitosan developed a more balanced Th1/Th2 response for both purified SeM and S. equi enzymatic extract proteins, although those induced by the pure antigen-entrapped particles were higher than the S. equi tested vaccines composed by total antigens entrapped in polymeric nanospheres.

Background: This work focuses on the development of atorvastatin utilizing zein, a natural, safe, and biocompatible polymer, as a nanosized formulation in order to overcome the poor oral bioavailability (12%) of the drug. Methods: Twelve experimental runs of atorvastatin-zein nanosphere formula were formulated by a liquid-liquid phase separation method according to custom fractional factorial design to optimize the formulation variables. The factors studied were: weight % of zein to atorvastatin (X 1), pH (X 2), and stirring time (X 3). Levels for each formulation variable were designed. The selected dependent variables were: mean particle size (Y 1), zeta potential (Y 2), drug loading efficiency (Y 3), drug encapsulation efficiency (Y 4), and yield (Y 5). The optimized formulation was assayed for compatibility using an X-ray diffraction assay. In vitro diffusion of the optimized formulation was carried out. A pharmacokinetic study was also done to compare the plasma profile of the atorvastatin-zein nanosphere formulation versus atorvastatin oral suspension and the commercially available tablet. Results: The optimized atorvastatin-zein formulation had a mean particle size of 183 nm, a loading efficiency of 14.86%, and an encapsulation efficiency of 29.71%. The in vitro dissolution assay displayed an initial burst effect, with a cumulative amount of atorvastatin released of 41.76% and 82.3% after 12 and 48 hours, respectively. In Wistar albino rats, the bioavailability of atorvastatin from the optimized atorvastatin-zein formulation was 3-fold greater than that from the atorvastatin suspension and the commercially available tablet. Conclusion: The atorvastatin-zein nanosphere formulation improved the oral delivery and pharmacokinetic profile of atorvastatin by enhancing its oral bioavailability.

Drugs with low aqueous solubility and high permeability (BCS class II) present a high proportion of all drugs. This study examines the critical issues regarding engineering of a nanosuspension tailored to increase drug dissolution rate and its transformation into dry powder suitable for tabletting. Nanosuspensions of celecoxib, a selective COX-2 inhibitor with low water solubility, were produced by the emulsion-diffusion method using three different stabilizers (Tween ® 80, PVP K-30 and SDS) and characterized by particle size analysis, dissolution testing, scanning electron microscopy imaging, differential scanning calorimetry and X-ray powder diffraction. Spray-dried nanosuspension was blended with microcrystalline cellulose, and compressed to tablets, and their tensile strength, porosity and elastic recovery of tablets were investigated. The selection of solvent and stabilizers is critical, firstly to achieve controlled crystallization and size, and secondly to increase the wettability of the hydrophobic drug. The crystalline nano-sized celecoxib alone or in tablets showed a dramatic increase of dissolution rate and extent compared to micronized. SEM images showed that the nanoparticle morphology was influenced by the choice of stabilizers. Celecoxib nanosuspension stabilized with PVP K-30 and SDS showed advantages over Tween ® 80 due to sticking of the dried product and unexpected changes observed on DSC curves. Markedly lower compaction forces are needed for nano-sized compared to micro-sized celecoxib to produce tablets of equal tensile strength.

Surface nanobubbles and micropancakes are two recent discoveries in interfacial physics. They are nanoscopic gaseous domains that form at the solid/liquid interface. The fundamental interest focuses on the fact that they are surprisingly stable to dissolution, lasting for at least 10-11 orders of magnitude longer than the classical expectation. So far, many articles have been published that describe various different nucleation methods and 'ideal' systems and experimental techniques for nanobubble research, and we are now at the stage where we can begin to investigate the fundamental questions in detail. In this topical review, we summarize the current state of research in the field and give an overview of the partial answers that have been proposed or that can be inferred to date. We relate nanobubbles and micropancakes, and we try to build a framework within which nucleation may be understood. We also discuss evidence for and against different aspects of nanobubble stability, as well as suggesting what 13 5. Typical sizes and distributions 13 5.1. Preferential sizes 13 5.2. Gas mobility 13 5.3. Contact angle 14 6. The depletion/precursor layer at the water/solid interface 15 6.1. A binary system (water and solid) 15 6.2. The long-range hydrophobic attraction 15 6.3. A tertiary system (water, gas and solid) 16 7. Micropancakes

The synthesis and characterization of spherical sugar-containing polymer brushes consisting of PS cores onto which chains of sugar-containing polymers have been grafted via two different techniques are described. Photopolymerization in aqueous dispersion using the functional monomer MAGlc and crosslinked or non-crosslinked PS particles covered with a thin layer of photo-initiator yielded homogeneous glycopolymer brushes attached to spherical PS cores. As an alternative, ATRP was used to graft poly-(N-acetylglucosamine) arms from crosslinked PS cores. Deprotection of the grafted brushes led to water-soluble particles that act as carriers for catalytically active gold nanoparticles. These glycopolymer chains show a high affinity to adsorb WGA whereas no binding to BSA or PNA could be detected.

Lipospheres are a drug encapsulation system composed of water dispersible solid microparticles of particle size between 0.01 and 100 m in diameter with a solid hydrophobic lipid core stabilized by a layer of phospholipid molecules embedded in their surface. The bioactive compound is dissolved or dispersed in the solid lipid matrix of the internal core. Since lipospheres were introduced in the beginning of the 1990s, they have been used for the delivery of multiple types of drugs by various routes of administration. Later, a self-assembling pro-nano lipospheres (PNL) encapsulation system was developed for oral drug delivery. Lipospheres have several advantages over other delivery systems, such as better physical stability, low cost of ingredients, ease of preparation and scale-up, high dispersibility in an aqueous medium, high entrapment of hydrophobic drugs, controlled particle size, and extended release of entrapped drug after administration, from a few hours to several days. This review article focuses on updated information on several aspects of lipospheres and PNL, including preparation techniques, physicochemical properties and in vitro evaluation methods. Additionally, it covers lipospheres and PNL utilization for oral, ocular, and parenteral delivery, with special attention to unique considerations and aspects for each route of administration.

We present a microfluidic cell-culture chip that enables trapping, cultivation and release of selected individual cells. The chip is fabricated by a simple hybrid glass-SU-8-PDMS approach, which produces a completely transparent microfluidic system amenable to optical inspection. Single cells are trapped in a microfluidic channel using mild suction at defined cell immobilization orifices, where they are cultivated under controlled environmental conditions. Cells of interest can be individually and independently released for further downstream analysis by applying a negative dielectrophoretic force via the respective electrodes located at each immobilization site. The combination of hydrodynamic cell-trapping and dielectrophoretic methods for cell releasing enables highly versatile single-cell manipulation in an array-based format. Computational fluid dynamics simulations were performed to estimate the properties of the system during cell trapping and releasing. Polystyrene beads and yeast cells have been used to investigate and characterize the different functions and to demonstrate biological compatibility and viability of the platform for single-cell applications in research areas such as systems biology.

Chitosan grafted with hydrophobic and hydrophilic groups initiates the formation of amphiphilic chitosan nanospheres. The molecular weight of mPEG plays an important role to control the particle size. As compared to mPEG 2000, which gives a bimodal nanosphere (w200, and w300 nm), mPEG 5000 initiates a monodispersed nanosphere with the smaller size (150 nm). In aqueous solution, the nanosphere surface is negatively charged resulting in a well dispersion in neutral to high pH but a significant precipitation in low pH. A model drug incorporation using lidocaine is successful when amphiphilic chitosan nanospheres were dissolved in good solvent followed by allowing mixing with drug solution before dialysis. The particle size of the drug incorporated chitosan is significantly increased, that is, from 100-150 nm to approximately 400-500 nm when the amount of incorporated lidocaine was about 0.68 mg per mg of lidocaine-loaded nanosphere.

The energy transport properties of plasmonic waveguides can be analyzed by solving the dispersion relation for surface plasmonpolaritons (SPPs). We use this approach to derive an approximate analytical expression for SPP propagation length when the waveguide is composed of linearly arranged metallic nanoparticles, while assuming that metal losses are small or partially compensated by gain. Applied to metal-dielectric (composite) nanospheres, the obtained expression allows us to optimize the performance of the waveguide and arrive at a number of practical design rules. Specifically, we show that SPP attenuation can be minimized at a certain interparticle distance for transverse modes, but gradually grows for both longitudinal and transverse modes with the increase of particle separation. We also show that the two basic methods of supplying gain to the system, i.e., embedding the particles into a gain medium or having a metal-gain composition for the particles, do not perform equally well and the former method is more efficient, but the way the two methods affect depends on the polarization of SPPs. To investigate the role of the nanoparticles' arrangement in determining SPP characteristics, we follow a purely numerical approach and consider a two-segment bent waveguide as an example. Analyzing the waveguide's transmission shows that it behaves in an oscillatory manner with respect to the angle between the two segments and is therefore higher for certain angles than for the others. This suggests that, in the design of waveguides with bends, careful attention needs to be paid in order to avoid bend angles that yield low transmission and to choose angles that give maximum transmission.

The entrapment of hematoporphyrin IX (Hp IX) in silica by means of a microemulsion resulted in silica spheres of 33 AE 6 nm. The small size, narrow size distribution and lack of aggregation maintain Hp IX silica nanospheres stable in aqueous solutions for long periods and permit a detailed study of the entrapped drug by different techniques. Hp IX entrapped in the silica matrix is accessed by oxygen and upon irradiation generates singlet oxygen which diffuses very efficiently to the outside solution. The Hp IX entrapped in the silica matrix is also reached by iron(II) ions, which causes quenching of the porphyrin fluorescence emission. The silica matrix also provides extra protection to the photosensitizer against interaction with BSA and ascorbic acid, which are known to cause suppression of singlet oxygen generation by the Hp IX free in solution. Therefore, the incorporation of Hp IX molecules into silica nanospheres increased the potential of the photosensitizer to perform photodynamic therapy.

Immune responses against the Leishmania antigens are not sufficient to protect against a leishmania challenge. Therefore these antigens need to be potentiated by various adjuvants and delivery systems. In this study, Poly (d,l-lactide-co-glycolide (PLGA) nanospheres as antigen delivery system and Quillaja saponins (QS) as immunoadjuvant have been used to enhance the immune response against autoclaved Leishmania major (ALM). PLGA nanospheres were prepared by a double-emulsion (W/O/W) technique. Particulate characteristics were studied by scanning electron microscopy and particle size analysis. Mean diameter for nanospheres loaded with ALM+QS was 294 ± 106 nm. BALB/c mice were immunized three times in 3-weeks intervals using ALM plus QS loaded nanospheres [(ALM+QS) PLGA ], ALM encapsulated with PLGA nanospheres [(ALM) PLGA ], (ALM) PLGA + QS, ALM + QS, ALM alone or PBS. The intensity of infection induced by L. major challenge was assessed by measuring size of footpad swelling. The strongest protection, showed by significantly (P < 0.05) smaller footpad, were observed in mice immunized with (ALM) PLGA . The (ALM+QS) PLGA group showed the least protection and highest swelling, while the (ALM) PLGA +QS, ALM+QS and ALM showed an intermediate protection with no significant difference. The mice immunized with ALM and ALM+QS showed the highest IgG2a/IgG1 ratio (P<0.01), followed by (ALM) PLGA +QS. The highest IFN-γ and lowest IL-4 production was seen in (ALM) PLGA +QS, ALM+QS groups. The highest parasite burden was observed in (ALM) PLGA +QS and (ALM+QS) PLGA groups. It is concluded that PLGA nanospheres as a vaccine delivery system could increase the protective immune responses, but QS adjuvant has a reverse effect on protective immune responses and the least protective responses were seen in the presence of this adjuvant.

A novel membrane-based lateral-flow immunodipstick assay was developed for the fast screening of aflatoxin B 2 (AFB 2) as a model compound in food samples. The detector reagent consisted of magnetic nanogold microspheres (MnGMs) with nano-Fe 2 O 3 particles as core and gold nanoparticles as shell, and bio-functionalized with monoclonal anti-AFB 2 antibodies. Manually spotted AFB 2-bovine serum albumin conjugates (AFB 2-BSA) and goat anti-mouse IgG on nitrocellulose membrane were used as test and control lines, respectively. As the major advantage, experimental results indicated that the visual detection limit (cutoff value) of the MnGM-based dipstick immunoassay with 0.9 ng/ml AFB 2 was about threefold lower compared to a conventional immunodipstick test using gold nanoparticles as detection reagent. Qualitative results (yes/no) could be obtained within 15 min without expensive equipment. The assay was evaluated with AFB 2 spiked or naturally contaminated samples (n = 8), including peanuts, hazelnuts, pistacia and almonds, receiving excellent correspondance with results from high performance liquid chromatography (HPLC). Most importantly, the assay gave no false negative results. By controlling the target antibody this assay can be easily extended for use with other food relevant toxins and thus represents a versatile detection method.

A reverse microemulsion method is reported for preparing monodispersed silica-coated gold (or silver) nanoparticles without the use of a silane coupling agent or polymer as the surface primer. This method enables a fine control of the silica shell thickness with nanometer precision. As compared to the Stöber method reported for direct silica coating, which can only coat large gold particles (∼50 nm in diameter) at low concentrations (<1.5 × 10 10 particles/mL), this new approach is capable of coating gold particles of a wide range of sizes (from 10 to 50 nm) at a much higher concentration (∼1.5 × 10 13 particles/mL). Moreover, it enables straightforward surface functionalization via cocondensation between tetraethyl orthosilicate and another silane with the desired functional groups. The functional groups introduced by this method are readily accessible and thus useful for various applications.

Magnetic hydrogel κ-carrageenan nanospheres were successfully prepared via water-in-oil (w/o) microemulsions combined with thermally induced gelation of the polysaccharide. The size of the nanospheres (an average diameter (∅) of about 50 and 75 nm) was modulated by varying the concentration of surfactant. The nanospheres contained superparamagnetic magnetite nanoparticles (∅8 nm), previously prepared by co-precipitation within the biopolymer. Carboxyl groups, at a concentration of about 4 mmol g-1, were successfully grafted at the surface of these magnetic nanospheres via carboxymethylation of the κ-carrageenan. The carboxylated nanospheres were shown to be thermo-sensitive in the 37-45 °C temperature range, indicating their potential as thermally controlled delivery systems for drugs and/or magnetic particles at physiological temperatures. Finally, preliminary results have been obtained for IgG antibody conjugation of the carboxylated nanospheres and the potential of these systems for bio-applications is discussed.

Currently, viruses are utilized as vectors for gene therapy, since they transport across cellular membranes, escape endosomes, and effectively deliver genes to the nucleus. The disadvantage of using viruses for gene therapy is their immune response. Therefore, nanospheres have been formulated as a nonviral gene vector by blending L-tyrosinepolyphosphate (LTP) with polyethylene glycol grafted to chitosan (PEG-g-CHN) and linear polyethylenimine (LPEI) conjugated to plasmid DNA (pDNA). PEG-g-CHN stabilizes the emulsion and prevents nanosphere coalescence. LPEI protects pDNA degradation during nanosphere formation, provides endosomal escape, and enhances gene expression. Previous studies show that LTP degrades within seven days and is appropriate for intracellular gene delivery. These nanospheres prepared by water-oil emulsion by sonication and solvent evaporation show diameters between 100 and 600 nm. Also, dynamic laser light scattering shows that nanospheres completely degrade after seven days. The sustained release of pDNA and pDNA-LPEI polyplexes is confirmed through electrophoresis and PicoGreen assay. A LIVE/DEAD cell viability assay shows that nanosphere viability is comparable to that of buffers. X-Gal staining shows a sustained transfection for 11 days using human fibroblasts. This result is sustained longer than pDNA-LPEI and pDNA-FuGENE 6 complexes. Therefore, LTP-pDNA nanospheres exhibit controlled transfection and can be used as a nonviral gene delivery vector.

Quantum dot-hydrotalcite layered nanoplatforms were successfully prepared following a one-pot synthesis. The process is very fast and a priori delamination of hydrotalcite is not a prerequisite for the intercalation of quantum dots. The novel materials were extensively characterized by X-ray diffraction, thermogravimetry, infrared spectroscopy, transmission electron microscopy, true color fluorescence microscopy, photoluminescence, and nitrogen adsorption. The quantum dot-hydrotalcite nanomaterials display extremely high stability in mimicking physiological media such as saline serum (pH 5.5) and PBS (pH 7.2). Yet, quantum dot release from the solid structure is noted. In order to prevent the leaking of quantum dots we have developed a novel strategy which consists of using tailor made double layered hydrotalcites as protecting shells for quantum dots embedded into silica nanospheres without changing either the materials or the optical properties.

Sub-micrometer-sized hollow tantalum oxide (Ta 2 O 5 ) spheres with tunable shell thickness and void size have been fabricated exploiting -diketone-functionalized polystyrene (PS) beads as sacrificial templates in a sol-gel process. First, a controlled precipitation of Ta 2 O 5 nanoparticles was carried out on the template surface by hydrolyzing tantalum ethoxide (Ta(OEt) 5 ) at room temperature, and subsequently, the polymer core was removed either via chemical treatment with toluene or calcination at 650°C. The thickness of the tantala shell precipitated on the PS core during the coating process was tuned between 100 and 142 nm by varying the concentration of tantala precursor in the reaction media. The obtained Ta 2 O 5 -coated PS particles and hollow microspheres were characterized by scanning electron microscopy, transmission electron microscopy, infrared spectroscopy, X-ray diffraction, and thermogravimetric analysis. Due to the unique optical and dielectric properties, these nanostructured materials are envisaged to be used in applications such as novel building blocks for the fabrication of advanced materials, surface coatings, catalysts, and drug delivery systems.

Hollow structures on the submicrometer scale (nm) are obtained with the assembly of polyelectrolytes according to the layer-by-layer (LbL) technique. Following the LbL procedure, polymers alginate and chitosan were alternatively adsorbed on a vesicular template made of didodecyldimethylammonium bromide (DDAB). Evidence for the removal of the vesicular template entrapped in the alginate/chitosan film is presented. The removal of the vesicular template was achieved through interactions between a nonionic surfactant (Triton X100) and the double-chained surfactant forming the vesicles. The application of this approach allowed the production of hollow nanospheres with a mild procedure, avoiding the use of strong acids or other extreme working conditions that can modify the shell integrity. The obtained nanostructures were characterized by means of dynamic light scattering (DLS), the ζ potential, and scanning electron microscopy (SEM). The SEM analysis demonstrated the presence, after the core removal process, of nanocapsules indistinguishable in size and shape from the parent core-shell system. The analysis of the surface charge of the hollow nanocapsules, after the core dissolution, by ζ potential measurements, indicates good aggregate stability. DLS measurements showed that the size of the nanocapsules is on the order of hundreds of nanometers. Moreover, the size of both the core-shell and the hollow particles did not appear to be perturbed by variations in temperature or ionic strength.