Human whole saliva (HWS) is a physiological secretion that performs a number of essential functions including protection of oral health, lubrication of mouth tissues and food pre-digestion. The complexity of the saliva proteome reflects the unique physiochemical functionality of this biofluid. In addition, saliva influences the transduction of aroma, taste and tactile (mouthfeel) perceptions through the interaction with food and beverages. This work, which is presented as two distinct contributions, reports on the extensive interfacial properties characterization (air-liquid) of saliva either alone or after exposure to tea polyphenols. This first part discusses the complex nature of the salivary film in terms of kinetic and viscoelastic (dilational surface rheology) properties. The formation of the surface film is analyzed with time and explained in terms of the progressively irreversible adsorption of protein fractions of larger molecular weight. The viscoelastic response to appropriate frequency modulation is modelled using a general mixed kinetic model, which satisfactorily reproduces the experimental data in better agreement than the traditional Lucassen diffusive model.

For the first time, electrocatalytic oxidation and selective determination of methadone (Mtd), as a long-acting opioid, in the presence of acetaminophen (Ac) has been investigated at a glassy carbon electrode modified with functionalized multi-walled carbon nanotubes. This simple and sensitive electrochemical sensor was fabricated through the drop-casting of functionalized multi-walled carbon nanotubes (fMWCNT) on the surface of a glassy carbon electrode (GCE). The electrocatalytic oxidations of Ac and Mtd are both individually and simultaneously investigated at the surface of the fMWCNT modified glassy carbon electrode (fMWCNT/MGCE) through using cyclic and differential pulse voltammetric studies. The fMWCNT/MGCE offered a considerable enhancement in the anodic peak current of Ac and Mtd associated with separating their overlapping voltammetric responses with potential difference of 290mV. The catalytic peak currents obtained from differential pulse voltammetry of Ac and Mtd increased linearly with their concentration at the ranges of 0.45-90.0μM and 0.5-100.0μM, respectively, and the detection limits for Ac and Mtd were sequentially 0.35μM and 0.28μM. Furthermore, this electrochemical sensor was successfully implemented for the quantitative determination of Ac and Mtd in human urine, saliva and pharmaceutical samples using standard addition method and the obtained results were found to be satisfactory.

Stable vesicles for efficient curcumin encapsulation, delivery and controlled release have been obtained by coating of liposomes with thin layer of newly synthesized chitosan derivatives. Three different derivatives of chitosan were obtained and studied: the cationic (by introduction of the stable, quaternary ammonium groups), the hydrophobic (by attachment of N-dodecyl groups) and cationic-hydrophobic one (containing both quaternary ammonium and N-dodecyl groups). Zeta potential measurements confirmed effective coating of liposomes with all these chitosan derivatives. The liposomes coated with cationic-hydrophobic chitosan derivative are the most promising curcumin carriers; they can easily penetrate cell membrane and release curcumin in a controlled manner. Biological studies indicated that such systems are non-toxic for murine fibroblasts (NIH3T3) while toxic toward murine melanoma (B16F10) cell line.

This article describes a rapid and facile method for manufacturing various size-controlled gel particles with utilizing inkjet printing technology. Generally, the size of droplets could be controlled by changing nozzle heads of inkjet printer, from which ink solution is ejected. However, this method uses drying process before gelling microparticles, and with that, the size of microparticles was easily controlled by only altering the concentration of ejected solution. When sodium alginate solution with various concentrations was ejected from inkjet printer, we found that the concentration of alginate solution vs. the volume of dried alginate particle showed an almost linear relationship in the concentration range from 0.1 to 3.0%. After dried alginate particles were soaked into calcium chloride solution, the size of microgel beads were obtained almost without increasing their size. The microparticles including various sizes of nanoparticles were easily manufactured by ejecting nanoparticle-dispersed alginate solution. The release of 25-nm sized nanoparticles from alginate microgel beads was finished in a relatively-rapid manner, whereas 100-nm sized nanoparticles were partially released from those ones. Moreover, most of 250-nm sized nanoparticles were not released from alginate microgel beads even after 24-h soaking. This particle fabricating method would enable the tandem drug delivery system with a combination of the release from nano and microparticles, and be expected for the biological and tissue engineering application.

Bone tissue engineering is a promising alternative method for treating bone loss by a combination of biomaterials and cells. In this study, we fabricated biocomposite scaffolds by blending chitosan (CS), alginate (Alg) and nano-silica (nSiO2), followed by freeze drying. The prepared scaffolds (CS/Alg, CS/Alg/nSiO2) were characterized by SEM, FT-IR and XRD analyses. In vitro studies such as swelling, biodegradation, biomineralization, protein adsorption and cytotoxicity were also carried out. The scaffolds possessed a well-defined porous architecture with pore sizes varying from 20 to 100μm suitable for cell infiltration. The presence of nSiO2 in the scaffolds facilitated increased protein adsorption and controlled swelling ability. The scaffolds were biodegradable and the addition of nSiO2 improved apatite deposition on these scaffolds. There was no significant cytotoxicity effect of these CS/Alg/nSiO2 scaffolds towards osteolineage cells. Thus, these results indicate that CS/Alg/nSiO2 scaffolds may have potential applications for bone tissue engineering.

In this study, stearic acid- and stearyl ferulate-based solid lipid nanoparticles containing trans-ferulic acid (SLN-FA and SLN-SF-FA, respectively), were prepared and characterized for loading efficiency, size and shape. In addition, by using rat brain microsomes, we evaluated in vitro the antioxidant activity of these formulations against three well known initiators of lipid peroxidation, such as AAPH, NADPH/ADP-Fe(3+) and SIN-1 which in turn generate the peroxyl and perferryl radicals as well as peroxynitrite, respectively. Commercially available FA and its ethyl ester (FAEE) were used as comparators. Both SLN-FA and SLN-SF-FA dose-dependently reduced lipid peroxidation induced by the three oxidants. Interestingly, SLN-SF-FA displayed greater efficacy (EC50) and potency (maximal activity) against AAPH- and NADPH/ADP-Fe(3+)-induced lipid peroxidation. Our results support the idea that this new formulations could facilitate the uptake of FA by the cells because of their lipophilic structure, thus increasing FA bioavailability. Furthermore, stearyl ferulate-based nanoparticles could prevent the degradation of FA entrapped on their structure, making FA almost entirely available to explicate its antioxidant power once released.

Macroporous cryogels imprinted with human serum albumin (HSA) have been prepared by copolymerization of 2-hydroxyethyl methacrylate with a functional co-monomer of N-methacryloyl-l-phenylalanine. The cryogels were used for the depletion of HSA from human serum. HSA-imprinted cryogels were prepared with gel fraction yields up to 90%, and their chemical structure, morphology and porosity were characterized by FTIR-spectroscopy, scanning electron microscopy, swelling studies and flow dynamics. Selective binding experiments were performed in the presence of competitive proteins like human transferrin and myoglobin. Albumin-imprinted cryogel column was optimized for fast protein liquid chromatography. Sodium-dodecyl sulfate polyacrylamide gel electrophoresis was used to show the efficiency of albumin depletion.

This study intended to develop nimodipine (NMD) nanocrystals with different sizes for oral administration and to investigate the relationship between dissolution and pharmacokinetics for NMD nanocrystals and Nimotop(®). NMD nanocrystals were prepared by combination of microprecipitation and high pressure homogenization and were further lyophilized. The particle size, morphology and aqueous solubility of the NMD nanocrystals were determined. With Nimotop(®) as the control, the dissolution rate was evaluated and the pharmacokinetic study was undertaken in beagle dogs. NMD nanocrystals with mean diameters of about 159.0, 503.0 and 833.3nm were prepared, respectively. The lyophilization didn't affect the particle sizes of the redispersed nanocrystals. The aqueous solubility was significantly improved and displayed a size-dependent manner. The nanocrystals exhibited lower dissolution patterns than Nimotop(®) under non-sink condition, but bioavailability of the two nanocrystals (159.0 and 833.3nm) was equivalent, about 2.6-fold higher than Nimotop(®). In conclusion, oral nanocrystal drug delivery system was a promising strategy in improving the oral bioavailability of poorly soluble or insoluble drugs. But we could not establish a favorable in vitro in vivo correlation for NMD nanocrystals and Nimotop(®) and thus the oral absorption mechanism of the NMD nanocrystals required further study.

Arachidyl chitosan (chitosan oligosaccharide-arachidic acid; CSOAA)-based self-assembled nanoprobes for magnetic resonance imaging (MRI) of neoplastic lesions was developed and evaluated in vitro. Diethylenetriaminepentaacetic dianhydride (DTPA) was conjugated to chitosan oligosaccharide (CSO) and Gd(3+) was chelated to the resulting ligand. DTPA conjugation and Gd(3+) chelation were confirmed primarily by Fourier transform infrared spectroscopy (FT-IR) and zeta potential measurement. A spherical nanoprobe of around 150nm mean diameter in the tested concentration range was formed in an aqueous environment by simple dissolution. The critical aggregation concentration (CAC) of the CSOAA-based nanoprobe was 3.86μg/ml, indicating its stability after dilution in body fluid. The nanoprobe had negligible toxicity in head and neck cancer cell lines (Hep-2 and FaDu cells). The amount of Cy5.5-labeled nanoprobe taken-up by cells, as observed by confocal laser scanning microscopy (CLSM), increased according to incubation time (up to 12h). A phantom study showed a T1-positive contrast-enhancing effect of the developed CSOAA-based nanoprobe, compared to that of the commercial formulation (Gd-DTPA; Magnevist). These results indicate that the CSOAA-based nanoprobe can be used for efficient MR imaging of neoplastic cells.

This study aimed to entrap bioprotective lactic acid bacteria in a sodium caseinate/sodium alginate aqueous two-phase system. Phase diagram at pH=7 showed that sodium alginate and sodium caseinate were not miscible when their concentrations exceeded 1% (w/w) and 6% (w/w), respectively. The stability of the caseinate/alginate two-phase system was also checked at pH values of 6.0 and 5.5. Lactococcus lactis subsp. lactis LAB3 cells were added in a 4% (w/w) caseinate/1.5% (w/w) alginate two-phase system at pH=7. Fluorescence microscopy allowed to observe that the caseinate-rich phase formed droplets dispersed in a continuous alginate-rich phase. The distribution of bacteria in such a system was observed by epifluorescence microscopy: Lc. lactis LAB3 cells stained with Live/Dead(®) Baclight kit™ were located exclusively in the protein phase. Since zeta-potential measurements indicated that alginate, caseinate and bacterial cells all had an overall negative charge at pH 7, the preferential adhesion of LAB cells was assumed to be driven by hydrophobic effect or by depletion phenomena in such biopolymeric systems. Moreover, LAB cells viability was significantly higher in the ternary mixture obtained in the presence of both caseinate and alginate than in single alginate solution. Caseinate/alginate phase separated systems appeared thus well suited for Lc. lactis LAB3 cells entrapment.