Bioaccessibility, Intestinal Absorption and Anti-Inflammatory Activity of Curcuminoids Incorporated in Avocado, Sunflower, and Linseed Beeswax Oleogels.

Beeswax oleogels (OGs), with a mechanical strength similar to pork backfat, were formulated with avocado (A), sunflower (S), and linseed (L) oils, applying a central composite design plus star point, and were evaluated as oral delivery vehicles of curcuminoids (OGACur, OGSCur, OGLCur). The incorporation of curcumin into the OG matrix significantly delayed both the formation of peroxides and conjugated trienes (K268 values), and the degradation rate of curcumin decreased with the increase of the oil polyunsaturated fatty acids (PUFA) content. The oil structuring did not affect the bioaccessibility of curcuminoids (>55% in all the OGs, regardless of the oil type), but it did reduce the release of fatty acids (~10%) during in vitro gastrointestinal digestion. The intestinal absorption, evaluated in Caco-2 cell monolayers, was higher for the micelle-solubilized curcumin from the digested OG than from unstructured oils, and it showed high anti-inflammatory potential by inhibiting the tumor necrosis factor-α (TNF-α) production compared to the positive control, both before and after the stimulation of ThP-1 cells with LPS. Regardless of the oil type, these beeswax-based OGs with gel-like behavior designed as fat replacers may be promising vehicles for the oral delivery of curcuminoids.

Bioaccessibility of chlorogenic acid and curcumin co-encapsulated in double emulsions with the inner interface stabilized by functionalized silica nanoparticles.

The aim of this study was to evaluate the bioaccessibility of chlorogenic acid (CA) and curcumin co-encapsulated in Pickering double emulsions (DEs) with the inner interface stabilized by hydrophobically modified silica nanoparticles with myristic acid (SNPs-C14) or tocopherol succinate (SNPs-TS). Both SNPs-C14 and SNPs-TS showed contact angles > 90°. Pickering W1/O emulsions were formulated with 4 % of both types of SNPs. Pickering DEs showed higher creaming stability (5-7 %, day 42) and higher CA encapsulation efficiency (EE; 80 %) than control DE. The EE of curcumin was > 98 % in all the DEs. CA was steadily released from Pickering DEs during digestion, achieving bioaccessibility values of 58-60 %. Curcumin was released during the intestinal phase (∼80 % bioaccessibility in all DEs). Co-loaded DEs showed similar bioaccessibility for CA and curcumin than single-loaded. SNPs-C14 and SNPs-TS were suitable to stabilize the W1:O interface of DEs as co-delivery systems of bioactive compounds with health-promoting properties.

Influence of the crystallinity on the physicochemical properties of spray-dried quercetin-inulin microparticles and their performance during in vitro digestion.

Encapsulation of quercetin (Q) with inulin (In) by spray-drying was performed applying a Box-Behnken design where the effect of the inlet air temperature, percentage of inulin crystallite dispersion and Q content were studied on the crystallinity index (CI). Three microparticle systems with CI between 2 % and 20 % (Q-In-2 %, Q-In-12 % and Q-In-20 %) were selected to study the CI effect on Q release during an in vitro digestion. The higher the CI of microparticles, the higher the encapsulation efficiency (76.4 %, Q-In-20 %). Surface quercetin was steadily released during the oral, gastric, and intestinal phases of the digestion. The CI of the microparticles did not influence the Q bioaccessibility values (23.1-29.7 %). The highest Q delivery occurred during the simulated colonic phase (44.4-66.4 %) due to the action of the inulinase. The controlled crystallization in spray-dried microparticles is a promising strategy for the designing of polyphenol-based microparticles with specific delivery properties.

Incorporation of hydroxytyrosol alkyl esters of different chain length as antioxidant strategy in walnut oil spray-dried microparticles with a sodium alginate outer layer.

Omega-3 rich vegetable oils, such as walnut oil, are gaining interest because of their health benefits. Synthetized homologous series of hydroxytyrosol alkyl esters (HTEs) with different alkyl chain lengths (C4-C18) were incorporated in purified walnut oil (PWO) spray-dried microparticles, designed with Capsul® (C) as encapsulating agent and sodium alginate (SA) as outer layer (PWO-C/SA). The encapsulation efficiency (>87%) and Tg of PWO-C/SA microparticles were not affected by the HTEs. The incorporation of HTE-C10 increased the melting point (185.0 ± 1.3 °C), decreasing the formation of Dimers + Polymers (1.12 ± 0.05% at day 35 of storage) and the crystallinity of the microparticles (>170 °C). The highest stability of PWO-C(HTE-C10)/SA suggests a specific location of HTE-C10 at the oil:water interface. The SA layer delayed the release of fatty acids during in vitro digestion. The incorporation of HTEs of medium chain length can be a suitable strategy to protect unsaturated oils encapsulated by spray-drying.

Encapsulation of Pomegranate Peel Extract (Punica granatum L.) by Double Emulsions: Effect of the Encapsulation Method and Oil Phase.

Pomegranate peel is an agro-industrial waste that can be used as source of punicalagin, a polyphenolic compound with several beneficial effects on health. Since, once extracted, punicalagin is prone to degradation, its encapsulation by double emulsions can be an alternative to protect the active compound and control its release. The aim of this investigation was to evaluate the feasibility of encapsulating pomegranate peel extract (PPE) in double emulsions using different types of oils (castor, soybean, sunflower, Miglyol and orange) in a ratio of 70:30 (oil:PPE) and emulsification methods (direct membrane emulsification and mechanical agitation), using polyglycerol polyricinoleate (PGPR) and Tween 80 as lipophilic and hydrophilic emulsifiers, respectively. Direct membrane emulsification (DME) led to more stable emulsions during storage. Droplet size, span values, morphology and encapsulation efficiency (EE) were better for double emulsions (DEs) prepared by DME than for mechanical agitation (MA). DEs formulated using Miglyol or sunflower oil as the oily phase could be considered as suitable food grade systems to encapsulate punicalagin with concentrations up to 11,000 mg/L of PPE.

Influence of the particle size of encapsulated chia oil on the oil release and bioaccessibility during in vitro gastrointestinal digestion.

Among vegetable oils, chia oil has been gaining interest in recent years due to its high linolenic acid content (ALA, 18:3 ω3). The aim of this work was to study the influence of the particle size of encapsulated purified chia oil (PCO) on the encapsulation efficiency and PCO release during in vitro digestion. PCO micro- and nano-sized particles with sodium alginate (SA) as an encapsulating agent (ME-PCO-SA and NE-PCO-SA) were designed by micro and nano spray-drying, respectively, applying a central composite plus star point experimental design. NE-PCO-SA showed a smaller particle size and higher encapsulation efficiency of PCO than ME-PCO-SA (0.16 µm vs. 3.5 µm; 98.1% vs. 92.0%). Emulsions (NE-PCO and ME-PCO) and particles (NE-PCO-SA and ME-PCO-SA) were subjected to in vitro static gastrointestinal digestion. ME-PCO and NE-PCO showed sustained oil release throughout the three phases of digestion (oral, gastric and intestinal phases), whereas the PCO release from ME-PCO-SA and NE-PCO-SA occurred mainly in the intestinal phase, showing the suitability of sodium alginate as an intestine-site release polymer. Nano-sized particles showed a significantly higher PCO release after in vitro digestion (NE-PCO-SA, 78.4%) than micro-sized particles (ME-PCO-SA, 69.8%), and also higher bioaccessibility of individual free fatty acids, such as C18:3 ω-3 (NE-PCO-SA, 23.6%; ME-PCO-SA, 7.9%), due to their greater surface area. However, when ME-PCO-SA and NE-PCO-SA were incorporated into yogurt, the PCO release from both particle systems after the digestion of the matrix was similar (NE-PCO-SA, 58.8%; ME-PCO-SA-Y, 61.8%), possibly because the calcium ions contained in the yogurt induced partial ionic gelation of SA, impairing the PCO release. Sodium alginate spray-dried micro and nanoparticles showed great potential for vehiculation of omega-3 rich oils in the design of functional foods.

Soybean oil organogelled emulsions as oral delivery systems of hydroxytyrosol and hydroxytyrosol alkyl esters.

Water-in-soybean oil organogelled emulsions (OGEs) were formulated as fat replacers and evaluated as delivery systems of hydroxytyrosol (HT, hydrophilic compound), hydroxytyrosol octanoate (HTC18, hydrophobic compound) and hydroxytyrosol decanoate (HTC10, with intermediate hydrophobicity and the highest antioxidant activity measured by conjugated autoxidizable triene assay). OGEs formulated with 55% of water and a ternary blend of candelilla wax, fully hydrogenated palm oil and monoacylglycerols showed mechanical properties similar to lard and solid-like behavior. The increase in the water content, together with a higher concentration of structuring agents in the oil phase, led to an increase in oil retention capacity and texture parameters. A slight desesterification of HTC10 and HTC18 was found during in vitro gastrointestinal digestion. The three bioactive compounds loaded in OGEs showed high bioaccessibility values (∼84%) at the end of digestion, regardless their chain length and hydrophobicity. These OGEs designed as fat replacers showed a great potential for vehiculation of both hydrophilic and lipophilic compounds.

Influence of the Location of Ascorbic Acid in Walnut Oil Spray-Dried Microparticles with Outer Layer on the Physical Characteristics and Oxidative Stability.

Purified walnut oil (PWO) microparticles with Capsul® (C, encapsulating agent), sodium alginate (SA) as outer layer and ascorbic acid (AA) as oxygen scavenger were obtained by spray drying using a three-fluid nozzle. AA was incorporated in the inner infeed (PWO-C(AA)/SA), in the outer infeed (PWO-C/SA(AA)) and in both infeed (PWO-C(AA)/SA(AA)). PWO-C(AA)/SA (4.56 h) and POW-C(AA)/SA(AA) (2.60 h) microparticles showed higher induction period than POW-C/SA(AA) (1.17 h), and lower formation of triacylglycerol dimers and polymers during storage (40 °C). Therefore, AA located in the inner infeed improved the oxidative stability of encapsulated PWO by removing the residual oxygen. AA in the SA outer layer did not improve the oxidative stability of encapsulated PWO since oxygen diffusion through the microparticles was limited and/or AA weakened the SA layer structure. The specific-location of AA (inner infeed) is a strategy to obtain stable spray-dried polyunsaturated oil-based microparticles for the design of foods enriched with omega-3 fatty acids.

IurV, Encoded by ORF VCA0231, Is Involved in the Regulation of Iron Uptake Genes in Vibrio cholerae.

The pathogen Vibrio cholerae has multiple iron acquisition systems which allow bacteria to exploit a variety of iron sources across the different environments on which it thrives. The expression of such iron uptake systems is highly regulated, mainly by the master iron homeostasis regulator Fur but also by other mechanisms. Recently, we documented that the expression of many of the iron-responsive genes is also modulated by riboflavin. Among them, the open reading frame VCA0231, repressed both by riboflavin and iron, encodes a putative transcriptional regulator of the AraC/XylS family. Nonetheless, the genes or functions affected by this factor are unknown. In the present study, a series of in silico analyses was performed in order to identify the putative functions associated with the product of VCA0231. The STRING database predicted many iron uptake genes as functional partners for the product of VCA0231. In addition, a genomic neighborhood analysis with the Enzyme Function Initiative tools detected many Pfam families involved in iron homeostasis genetically associated with VCA0231. Moreover, a phylogenetic tree showed that other AraC/XylS members known to regulate siderophore utilization in bacteria clustered together and the product of VCA0231 localized in this cluster. This suggested that the product of VCA0231, here named IurV, is involved in the regulation of iron uptake processes. RNAseq was performed to determine the transcriptional effects of a deletion in VCA0231. A total of 52 genes were overexpressed and 21 genes were downregulated in response to the iurV deletion. Among these, several iron uptake genes and other iron homeostasis-related genes were found. Six gene ontology (GO) functional terms were enriched in the upregulated genes, of which five were related to iron metabolism. The regulatory pattern observed in the transcriptomics of a subset of genes was independently confirmed by quantitative real time PCR analysis. The results indicate that IurV is a novel regulator of the AraC/XylS family involved in the repression of iron uptake genes. Whether this effect is direct or indirect remains to be determined.

Effect of Adding Curcumin on the Properties of Linseed Oil Organogels Used as Fat Replacers in Pâtés.

Beeswax-based organogels were formulated with linseed oil and curcumin according to a statistical design to increase the oxidative stability of spreadable meat products (pâté) where these organogels (OGCur) were incorporated as fat substitutes. The organogels obtained under optimal conditions (9.12% beeswax, 0.54% curcumin) showed a mechanical strength similar to pork backfat determined by back extrusion and high oil binding capacity (OBC; over 90%). The incorporation of curcumin at this concentration did not lead to any change in the arrangement of the crystal network, OBC, and mechanical, thermal, or rheological properties of the organogels. Beeswax organogels with and without curcumin, with a ß' orthorhombic subcell structure, showed a predominant elastic behavior and a melting event wider and shifted to lower temperatures than pure beeswax, suggesting a plasticizer effect of the oil in the wax crystals. The oxidative stability of the organogels under accelerated oxidation conditions increased due to the incorporation of curcumin. A decrease in the curcumin content was found from day 4 at 60 °C, together with a significantly lower formation of both peroxides and malonaldehyde. When pork backfat was partially or totally replaced by OGCur in pâtés, a noticeable protective effect of curcumin against lipid oxidation was found during chilled storage.

Effect of lyophilization on the physicochemical and rheological properties of food grade liposomes that encapsulate rutin.

The potential use of liposomes as carriers for food active ingredients can be limited by their physical and chemical instabilities in aqueous dispersions, especially for long-term storage. Lyophilization, a process commonly used in the food industry, can also be applied to stabilize and preserve liposomes and to extend their shelf-life. In this work, liposomes with potential use for designing functional foods were prepared with soy phospholipids and rutin. Homogenization and ultrasound were used for particle size reduction. Liposomal stability was evaluated by Dynamic Light Scattering, microscopy and rheological properties. Spherical and unilamellar liposomes were obtained in this work. Zeta potential (ξ = values were around -40 mV), which indicates a great suspension stability even for more than 30 days of storage. Rutin exerted a protective effect by both preventing damage to the liposome bilayer and maintaining the spherical structure after 56 days of storage. Lyophilization caused an increase in the size of the vesicles, reaching sizes around 419 nm and aggregation of vesicles with probably structural damage after 21 storage days. However, it helped to keep the rutin encapsulated (81.9%) for longer time, when compared to refrigerated liposomes. Rheological measurements showed, in general, that the power law model fitted most of the experimental results and dynamic rheological tests showed a sol-gel phase transition between 35 and 45 °C. Lyophilization caused a significant change in all evaluated rheological parameters. For the in vitro release tests, the liposomal bilayer acted as a barrier for the rutin release to the food simulating medium; therefore, the release rate of the antioxidant from the rutin encapsulated liposome was slow compared to the free rutin release rate.

Influence of gelation on the retention of purple cactus pear extract in microencapsulated double emulsions.

A purple cactus pear (Opuntia ficus-indica) extract (CP) was encapsulated in double emulsions (DE) gelled with gelatin (DE-CP-G) and with gelatin and transglutaminase (DE-CP-GT), as well as in a DE with a liquid external aqueous phase (DE-CP), in order to study the retention of betanin as colorant agent. Both gelled DEs showed a predominantly elastic behavior, in contrast with DE-CP. The degradation rate constant of betanin was significantly higher in DE-CP-GT (90.2 x 10-3 days-1) than in DE-CP-G (11.0 x 10-3 days-1) and DE-CP (14.6 x 10-3 days-1) during cold-storage (4 °C). A shift towards yellow color was found in all the systems during cold-storage (4 °C) and after thermal treatment (70°C/30 min), especially in DE-CP-GT, denoting a higher degradation of betanin. Betalamic acid, cyclo-Dopa 5-O-ß-glucoside, 17-decarboxy-betanin and neobetanin were identified by UHPLC-MS/MS as degradation products of betanin.

Role of maltodextrin and inulin as encapsulating agents on the protection of oleuropein during in vitro gastrointestinal digestion.

Olive leaves extract (OLE) was spray-dried with maltodextrin (MD) or inulin (IN) to study the evolution of oleuropein (OE) during in vitro gastrointestinal digestion, its bioaccessibility and potential bioavailability. In the case of OLE-MD, OE was partially degraded in gastric and intestinal conditions; whereas in OLE-IN, OE was released under gastric conditions and partially degraded under intestinal conditions. In both cases, the encapsulation of OLE led to higher OE contents at the end of digestion, compared with non-encapsulated OLE, suggesting a protective role of the polysaccharides by the formation of non-covalent polysaccharides-OE complexes. OE bioaccessibility was ten times higher (p ≤ 0.05) in OLE-MD and OLE-IN than in non-encapsulated OLE. However, OE potential bioavailability, evaluated by tangential filtration, was not detected. Encapsulation technology and the encapsulant agent used may determine the release of the encapsulated compounds at a specific-site and their effect on health.

Influence of the Physical State of Spray-Dried Flavonoid-Inulin Microparticles on Oxidative Stability of Lipid Matrices.

The effect of the physical state of flavonoid-inulin microparticles (semi-crystalline/amorphous) on the oxidative stability of lipid matrices was studied. Epicatechin (E) and quercetin (Q) microparticles with inulin were formulated at two infeed temperatures (15 °C and 90 °C) by spray drying. X-ray diffraction analyses showed that flavonoid-inulin microparticles obtained at feed temperature of 15 °C were semi-crystalline (E-In-15, 61.2% and Q-In-15, 60%), whereas those at 90 °C were amorphous (Q-In-90, 1.73 and Q-In-90 2.30%). Semi-crystalline state of flavonoid-inulin microparticles enhanced the EE (68.8 and 67.8% for E and Q, respectively) compared to amorphous state (41.6 and 51.1% for E and Q, respectively). However, amorphous Q-microparticles showed the highest antioxidant activity both in methyl linoleate and sunflower oil, increasing the induction period and decreasing the polar compounds and polymer triglyceride formation during long-term oxidation study. Therefore, the physical state of spray-dried flavonoid-inulin microparticles may determine their antioxidant activity in lipid matrices.

Double emulsions with olive leaves extract as fat replacers in meat systems with high oxidative stability.

Double emulsions (DE) with a healthy oil blend as lipid phase and an olive leave extract (OLE) encapsulated in the internal aqueous phase (DE/OLE) were incorporated as fat replacers in meat systems, in order to improve both the lipid profile and the oxidative stability. After 14 days of storage at 4 °C, DE/OLE showed good physical stability (90% of globule population was still below 10 µm diameter), and high antioxidant capacity (over 80%), longer than time required for this type of food ingredients. A high correlation was found between the remaining oleuropein content and the antioxidant capacity in both meat systems with DE/OLE (MS-DE/OLE) and meat systems with the oil blend as liquid oil and non-encapsulated OLE (MS-L/OLE). MS-DE/OLE were technologically feasible and showed higher retention of oleuropein (69%), oxidative stability and antioxidant capacity at 60 °C for 7 days than MS-L/OLE, where oleuropein was almost depleted. The encapsulation of OLE in DE could be a suitable strategy to avoid lipid oxidation in meat systems with healthier lipid profile.

Evolution of the phenolic compounds profile of olive leaf extract encapsulated by spray-drying during in vitro gastrointestinal digestion.

An olive leaf extract (OLE) was microencapsulated with sodium alginate (SA) by spray-drying to study the evolution of oleuropein (ORP) during in vitro gastrointestinal digestion, and its bioaccessibility and potential bioavailability from OLE and OLE-SA microparticles. Secoiridoids, flavonoids, simple phenols, oleosides and elenolic acid were identified in OLE. OLE/SA ratio 1:1.6 and inlet air temperature 135 °C were the optimal conditions for OLE-SA microparticles. ORP (70%) from OLE was degraded during gastric digestion, giving hydroxytyrosol and ORP-aglycone, whereas only the superficial ORP was released from microparticles. The remaining ORP from OLE was degraded under intestinal conditions, leading to oleosides; whereas alginate was swollen and disintegrated, releasing the ORP (90% of encapsulated ORP). ORP from both OLE and microparticles was degraded to hydroxytyrosol under colonic conditions. Encapsulation of OLE allowed the protection of ORP under gastric conditions and its controlled release at intestinal conditions, and higher bioaccessibility (58%) and potential bioavailability (20%).

Effect of spray-drying with organic solvents on the encapsulation, release and stability of fish oil.

Fish-oil (FO) was encapsulated with hydroxypropylcelullose (HPC) by conventional spray-drying with water (FO-water) and solvent spray-drying with ethanol (FO-EtOH), methanol (FO-MeOH) and acetone (FO-Acet) in order to study the effect of the solvent on the encapsulation efficiency (EE), microparticle properties and stability of FO during storage at 40 °C. Results showed that FO-Acet presented the highest EE of FO (92.0%), followed by FO-EtOH (80.4%), FO-MeOH (75.0%) and FO-water (71.1%). A decrease of the dielectric constant increased the EE of FO, promoting triglyceride-polymer interactions instead of oil-in-water emulsion retention. FO release profile in aqueous model was similar for all FO-microparticles, releasing only the surface FO, according to Higuchi model. Oxidative stability of FO significantly improved by spray-drying with MeOH, both in surface and encapsulated oil fractions. In conclusion, encapsulation of FO by solvent spray-drying can be proposed as an alternative technology for encapsulation of hydrophobic molecules.

Double emulsions as potential fat replacers with gallic acid and quercetin nanoemulsions in the aqueous phases.

The development of fat replacers to obtain healthier/functional foods is a constant challenge. With this aim, double emulsions (DE) with a blend of olive, linseed and fish oils as oil phase were developed. To prevent the oxidation of these oils, gallic acid and quercetin were incorporated in the internal and the external aqueous phase (W2), respectively, according to a factorial design. Considering the low solubility of quercetin in water, it was included in O/W nanoemulsions (QN), thus being freely dispersible in W2. The antioxidant activity in DE was attributed to QN, which significantly improved the oxidative stability of DE/QN. Furthermore, DE/QN showed good physical stability with a limited coalescence during storage at 4 °C for 28 days, significantly longer than time usually required for food ingredients. Therefore, DE/QN could be used as potential fat replacer in a variety of food formulations, providing blends of fatty acids consistent with dietary recommendations.

Incorporation of liposomes containing squid tunic ACE-inhibitory peptides into fish gelatin.

BACKGROUND: Hydrolysates from collagen of jumbo squid (Dosidicus gigas) tunics have shown excellent angiotensin I-converting enzyme (ACE)-inhibitory activity. However, peptides directly included in food systems may suffer a decrease in activity, which could be minimized by loading them into nanoliposomes. RESULTS: A fraction of peptides with molecular weights <1 kDa obtained from hydrolyzed squid tunics, with reasonably high ACE-inhibitory activity (half-maximal inhibitory concentration IC50 = 0.096 g L(-1)), was encapsulated in phosphatidylcholine nanoliposomes. The peptide concentration affected the encapsulation efficiency and the stability of the resulting liposomes, which remained with a high zeta potential value (-54.3 mV) for at least 1 week at the most suitable peptide concentration. The optimal peptide concentration was established as 1.75 g L(-1). Liposomes obtained with this peptide concentration showed an encapsulation efficiency of 53%, a zeta potential of -59 mV, an average diameter of 70.3 nm and proved to be stable in the pH range 3-7 at 4 °C. CONCLUSION: Liposomes containing ACE-inhibitory peptides were incorporated in fish gelatin without detriment to the rheological properties and thermal stability of the resulting cold-induced gel. The ACE-inhibitory activity of the peptide fraction, which was not affected by the encapsulation process, conferred the bioactive potential to the nanoliposome-containing gelatin gel.