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Electrospun xanthan polysaccharide nanofibers (X) were developed as an encapsulation and delivery system of the poorly absorbed polyphenol compounds, gallic acid (GA) and (-)-epigallocatechin gallate (EGCG). Scanning electron microscopy was used to characterize the electrospun nanofibers, and controlled release studies were performed at pH 6.5 and 7.4 in saline buffer, suggesting that the release of polyphenols from xanthan nanofibers follows a non-Fickian mechanism. Furthermore, the X-GA and X-EGCG nanofibers were incubated with Caco-2 cells, and the cell viability, transepithelial transport, and permeability properties across cell monolayers were investigated. An increase of GA and EGCG permeability was observed when the polyphenols were loaded into xanthan nanofibers, compared to the free compounds. The observed in vitro permeability enhancement of GA and EGCG was induced by the presence of the polysaccharide nanofibers, which successfully inhibited efflux transporters, as well as by tight junctions opening.

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Xanthan-Chitosan (X-Ch) polysaccharides nanofibers were prepared using electrospinning processing as an encapsulation and delivery system of curcumin (Cu). The X-Ch-Cu nanofibers remained stable in aqueous HBSS medium at pH 6.5 and pH 7.4, mainly due to the ability of oppositely charged xanthan-chitosan polyelectrolytes to form ionically associated electrospun nanofibers. The xanthan-chitosan-curcumin nanofibers were incubated with Caco-2 cells, and the cell viability, transepithelial transport and permeability properties across cell monolayers were investigated. After 24 h of incubation, the exposure of Caco-2 cell monolayers to X-Ch-Cu nanofibers resulted in a cell viability of ∼80%. A 3.4-fold increase of curcumin permeability was observed when the polyphenol was loaded into X-Ch nanofibers, compared to the free curcumin. This increased in vitro transepithelial permeation of curcumin without compromising cellular viability was induced by interactions upon contact between the nanofibers and the Caco-2 cells, leading to the opening of the tight junctions. The results obtained revealed that X-Ch nanofibers can be used for oral delivery applications of poorly water-soluble compounds at the gastrointestinal tract.

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Stable and low-cost multiplexed drug sensitivity assays using small volumes of cells or tissue are in demand for personalized medicine, including patient-specific combination chemotherapy. Spatially defined projected light photopolymerization of hydrogels with embedded active compounds is introduced as a flexible and cost-efficient method for producing multiplexed dosing assays. The high spatial resolution of light projector technology defines multiple compound doses by the volume of individual compound-embedded hydrogel segments. Quantitative dosing of multiple proteins with a dynamic range of 1-2 orders of magnitude is demonstrated using fluorescently labeled albumins. The hydrogel matrix results from photopolymerization of low-cost poly(ethylene glycol) diacrylates (PEGDA), and tuning of the PEGDA composition enables fast complete dosing of all tested species. Dosing of hydrophilic and hydrophobic compounds is demonstrated using two first-line chemotherapy regimens combining oxaliplatin, SN-38, 5-fluorouracil, and folinic acid, with each compound being dosed from a separate light-defined hydrogel segment. Cytotoxicity studies using a colorectal cancer cell line show equivalent effects of dissolved and released compounds. Further control of the dosing process is demonstrated by liposomal encapsulation of oxaliplatin, stable embedding of the liposomes in hydrogels for more than 3 months, and heat-triggered complete release of the loaded oxaliplatin.

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A novel molecular beacon (a nanomachine) is constructed that can be actuated by a radio frequency (RF) field. The nanomachine consists of the following elements arranged in molecular beacon configuration: a gold nanoparticle that acts both as quencher for fluorescence and a localized heat source; one reporter fluorochrome, and; a piece of DNA as a hinge and recognition sequence. When the nanomachines are irradiated with a 3 GHz RF field the fluorescence signal increases due to melting of the stem of the molecular beacon. A control experiment, performed using molecular beacons synthesized by substituting the gold nanoparticle by an organic quencher, shows no increase in fluorescence signal when exposed to the RF field. It may therefore be concluded that the increased fluorescence for the gold nanoparticle-conjugated nanomachines is not due to bulk heating of the solution, but is caused by the presence of the gold nanoparticles and their interaction with the RF field; however, existing models for heating of gold nanoparticles in a RF field are unable to explain the experimental results. Due to the biocompatibility of the construct and RF treatment, the nanomachines may possibly be used inside living cells. In a separate experiment a substantial increase in the dielectric losses can be detected in a RF waveguide setup coupled to a microfluidic channel when gold nanoparticles are added to a low RF loss liquid. This work sheds some light on RF heating of gold nanoparticles, which is a subject of significant controversy in the literature.

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We report on the surface characterization, functionalization, and application of stable water suspensions of novel surface active maghemite nanoparticles (SAMNs), characterized by a diameter of 11 ± 2 nm and possessing peculiar colloidal properties and surface interactions. These features permitted the acquisition of titration curves and aqueous UV-vis spectra and suggested a role played by surface under-coordinated iron atoms. This new class of nanoparticles was obtained through an easy, inexpensive, one-step, green procedure and functionalized with ligands of high biotechnological interest, such as biotin and avidin, by simple incubation in aqueous solution. Bound avidin was determined by measuring the disappearance of free avidin absorbance at 280 nm, as a function of increasing nanoparticle concentration, showing the presence of 10 ± 3 avidin molecules per nanoparticle. The biological activity of the SAMN@avidin complex was evaluated and the number of available biotin binding sites was determined, using biotinyl-fluorescein as a probe, showing that each bound avidin molecule is able to bind 2.8 ± 0.8 biotin molecules, confirming the maintenance of biological activity and excellent binding capacity of the SAMN@avidin complex. Furthermore a Langmuir isotherm model was used to describe the biomolecule specific monolayer adsorption onto the particle surface, and in the case of avidin, the maximum adsorption capacity was 100 ± 27 µg avidin/mg SAMN, whereas the binding constant is 45.18 µL µg(-1). The SAMN@avidin complex was characterized by UV-vis spectroscopy, quartz crystal microbalance, FTIR spectroscopy, and transmission electron microscopy. Finally, SAMN@avidin was applied for the large scale purification of recombinant biotinylated human sarco/endoplasmic reticulum Ca(2+)-ATPase (hSERCA-2a), expressed by Saccharomyces cerevisiae. The protein was magnetically purified, and about 500 µg of a 70% pure hSERCA-2a were recovered from 4 L of yeast culture, with a purification yield of 64%.

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