Development, characterization, and evaluation of the antioxidant activity of nanocarriers based on surfactant swollen micelles that encapsulate essential oils.

This work aimed to compare the performance of two relatively underexplored methods for the swollen micelles (SMs) production as nanocarriers for essential oils (EOs). Origanum vulgare and Thymus vulgaris EOs were examined. The first method (SMs-1), involved a self-assembly process, while the second one (SMs-2), employed titration operation of an emulsion into a surfactant solution for SMs formation. Tween 80 and ethanol were used as surfactant and co-surfactant, respectively. The solubilization kinetics and the saturation concentration of EOs were determined. Particle size (measured by DLS) and encapsulation efficiency (EE) were the control parameters assessed, along with the EOs-loaded SMs' stability during 30 days of storage. Additionally, the EOs-loaded SMs' morphology was analyzed using atomic force microscopy (AFM). Finally, the antioxidant activity through the ABTS+ radical scavenging and the reducing power of EOs encapsulated in SMs was determined. The results showed that the solubilization of EOs in SMs was a rapid process with high EE. EOs-loaded SMs-2 systems exhibited greater colloidal stability and higher EE compared to EOs-loaded SMs-1 systems, showing smaller and more homogeneous particle sizes. Moreover, EOs-loaded SMs-2 systems maintained constant EE throughout the storage period. AFM imaging confirmed the rounded and heterogeneous morphology of EOs-loaded SMs-1 and the smaller, more homogeneous, and spherical morphology of EOs-loaded SMs-2. EOs-loaded SMs-2 showed high ABTS+ radical scavenging and reducing power when encapsulated in SMs. In conclusion, the SMs-2 method emerged as an effective approach for producing efficient nanocarriers for EOs, signifying a promising path for future developments in antioxidant delivery systems.

Bioactive compounds: Application of albumin nanocarriers as delivery systems.

Enriched products with bioactive compounds (BCs) show the capacity to produce a wide range of possible health effects. Most BCs are essentially hydrophobic and sensitive to environmental factors; so, encapsulation becomes a strategy to solve these problems. Many globular proteins have the intrinsic ability to bind, protect, encapsulate, and introduce BCs into nutraceutical or pharmaceutical matrices. Among them, albumins as human serum albumin (HSA), bovine serum albumin (BSA), ovalbumin (OVA) and α-lactalbumin (ALA) are widely abundant, available, and applied in many industrial sectors, becoming promissory materials to encapsulate BCs. Therefore, this review focuses on researches about the main groups of natural origin BCs (namely phenolic compounds, lipids, vitamins, and carotenoids), the different types of nanostructures based on albumins to encapsulate them and the main fields of application for BCs-loaded albumin systems. In this context, phenolic compounds (catechins, quercetin, and chrysin) are the most extensively BCs studied and encapsulated in albumin-based nanocarriers. Other extensively studied subgroups are stilbenes and curcuminoids. Regarding lipids and vitamins; terpenes, carotenoids (ß-carotene), and xanthophylls (astaxanthin) are the most considered. The main application areas of BCs are related to their antitumor, anti-inflammatory, and antioxidant properties. Finally, BSA is the most used albumin to produced BCs-loaded nanocarriers.

In vitro gastrointestinal digestion and cytotoxic effect of ovalbumin-conjugated linoleic acid nanocomplexes.

The aim of this work was to examine the behavior of conjugated linoleic acid (CLA) delivery systems based on ovalbumin (OVA) and their derived nanoparticles (OVAn1 and OVAn2), under static in vitro gastrointestinal digestion model. In addition, potential cytotoxic effect of these inclusion complexes on a human colon cancer cell line (HT-29) was evaluated. OVA was resistant to gastric and intestinal digestion, while OVA nanoparticles were very susceptible to digestive enzymes hydrolysis. Particle size distribution (PDS) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for OVA evidenced the presence of a protein fragment of similar size after simulated digestive process. Conversely, for nanoparticles, partial and total hydrolysis in gastric and intestinal phases, respectively, was evidenced. After in vitro gastrointestinal digestion, released CLA (RCLA) was assayed. In case of OVA, as CLA carrier, RCLA was 37%, while for OVA nanoparticles, lower RCLA values (~10-20%) were obtained. From cytotoxic assays, it was observed that CLA molecule was responsible for cell death, whereas OVA or their derived nanoparticles were not cytotoxic on HT-29 cells. On the other hand, flow cytometry analysis revealed that main death mechanism for CLA, and their inclusion complexes was apoptosis. OVA-CLA and OVAn1-CLA inclusion complexes displayed the highest potential cytotoxic activity and apoptotic index. Information derived from this work could be relevant for the design of CLA delivery systems as promising nanosupplements for production of new functional and excipient foods for both prevention and control of colon cancer.

Physiological and morphological responses of green microalgae Chlorella vulgaris to silver nanoparticles.

The toxic effects of silver nanoparticles (AgNPs) on the physiology and morphology of the green microalga Chlorella vulgaris were studied. AgNPs were characterized by particle size distribution, ζ potential measurement, and atomic force microscopy (AFM). Chlorella vulgaris was exposed to 90-1440 µg/L of AgNPs range in Bold's Basal Medium for 96 h. The inhibition of algae growth rate and changes in the concentrations of chlorophyll-a, chlorophyll-b, pheophytin, and carotenoids was determined at the beginning and end of the trial. Cell diameter and volume, carbohydrate, total lipids, and protein content were also determined. Our data strongly suggest that the toxic effects of the AgNPs resulted in concentration and time-dependent. AgNPs altered C. vulgaris growth kinetics and cell metabolism expressed in photosynthetic pigments and biochemical composition. Our study confirmed the cytotoxicity of AgNPs through the algal growth inhibition with an EC50 value of 110 µg/L. Also, changes of chlorophyll-a, chlorophyll-b, pheophytin, and carotenoids concentrations were observed associated with a color shift from green to pale brown of algae cultures exposed to AgNPs for 96 h. Furthermore, algae cell concentration, diameter, and volume, plus total lipid, protein, and carbohydrates contents in the presence of AgNPs, were significantly altered compared to untreated cells. In synthesis, this study highlighted AgNPs toxic effects on morphological and physiological traits of C. vulgaris and warns about possible impacts on energy flow and aquatic food web structure, and on the transfer efficiency of energy to higher trophic levels.

Simulated gastrointestinal digestion of inclusion complexes based on ovalbumin nanoparticles and conjugated linoleic acid.

The objective of this work was to obtain and characterize conjugated linoleic acid (CLA) delivery systems based on ovalbumin nanoparticles (OVAn1 and OVAn2) and to study their behaviour under a standardised static in vitro digestion model. OVAn1 and OVAn2 were obtained by heat treatment (85 °C, 5 min, pH 11.35 and 7.5, respectively). OVAn1 and OVAn2 had hydrodynamic diameters of 24.63 ± 0.04 and 92.0 ± 0.2 nm, respectively, showing no significant differences in ζ potential values (p < 0.05) at pH 7.0. CLA nanocomplexes were examined in terms of size and ζ potential at pH 3.0 and 7.0, highlighting that binding of CLA caused an increase in size for OVA and both OVA nanoparticles. Morphological characterization was performed by confocal laser scanning microscopy (CSLM) finding that OVA and OVA nanoparticles had a circular shape. Also, the CLA encapsulation efficiency (EE) for OVA and OVA nanoparticles was studied, yielding EE values greater than 97% for all systems. Finally, systems were assayed for a standardized in vitro gastrointestinal digestion model considering gastric and intestinal steps. Macroscopic appearance, CSLM images and quantification of CLA retention by HPLC were evaluated after digestion. All the systems showed the formation of macroscopic aggregates both in gastric and intestinal phases, which generated a visible precipitate. In all systems, CSLM confirmed the presence of numerous undefined-form aggregates. Finally, high CLA retention (around 99%) was found for native protein and nanoparticles.

Formation and characterization of self-assembled bovine serum albumin nanoparticles as chrysin delivery systems.

Chrysin (5,7-dihydroxyflavone) (Chrys) is a natural flavone extracted from many plants, and it has been proposed as a bioactive agent for cancer therapy. Nevertheless, its use is limited mainly due to its poor water solubility. Bovine serum albumin (BSA) is a water soluble, biocompatible and non-toxic protein with a promising application in lipophilic bioactive compound delivery. Moreover, BSA is heat sensitive, feature that could be used for producing self-assembled nanoparticle with tailor-made properties. In this contribution, we studied the formation of BSA nanoparticles (BSAnp) by thermal treatment at different conditions of temperature (70 °C/5 min and 85 °C/5 min), protein concentration (1.0-4.0%wt.) and aqueous medium pH values (9.0 and 11.0) in which it is known that BSA is found in different unfolded conformations. Binding of Chrys dissolved in dimethyl sulfoxide (DMSO) was studied by fluorescence titration experiments. Characterization of Chrys-loaded and unloaded BSAnp was performed in phosphate buffered saline (PBS) pH 7.4 by applying a set of complementary techniques: dynamic light scattering (DLS), size exclusion fast protein liquid chromatography (SEC-FPLC) and transmission electron microscopy (TEM). Different populations of BSAnp were obtained, which showed different diameters in the range of 1328 nm, ζ potentials around -10.0 mV, molecular weight in the range of 400-1000 kDa and spherical shape. Chrys encapsulation efficiency (EE. %) was also determined, and values between 44-84% were obtained, which mainly depended on the mode of Chrys binding and physicochemical BSAnp properties. Results highlight the ability of self-assembled BSAnp for Chrys vehiculization in an aqueous medium which could found potential application in antitumor therapies.