Enhancing stability and bioavailability of sulforaphene in radish seed extracts using nanoemulsion made with high oleic sunflower oil.

The effect of nanoemulsions on the stability and bioavailability of sulforaphene (SFEN) in radish seed extract (RSE) was investigated. Four types of oil were used as lipid ingredients of the nanoemulsions: soybean, high oleic acid sunflower, coconut, and hydrogenated palm oils. SFEN in RSE nanoemulsions showed greater stability to temperature, acid, and alkaline conditions than SFEN in RSE suspended in water (RSE-S). Particularly under alkaline conditions, the half-life of SFEN in the nanoemulsion with high oleic sunflower oil (RSE-HOSO) was 8 times longer than that of RSE-S. Furthermore, in the pharmacokinetics study, it was observed that AUC0-8 increased and oral clearance (CL/F) decreased significantly in rats orally administered RSE-HOSO compared with RSE-S (p < 0.05). This study indicates that the type of oil used in nanoemulsions affects the stability and bioavailability of SFEN in RSE. These results may provide a guideline for the development of functional foods containing RSE. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01304-2.

Applications and perspectives of polyphenol-loaded solid lipid nanoparticles and nanostructured lipid carriers for foods.

Imbalanced nutrition in modern society is one of the reasons for disorders, such as cancer, cardiovascular disease, and diabetes, which have attracted the interest in bioactives (particularly polyphenols) to assist in the balanced diet of modern people. Although stability can be maintained during preparation and storage, the ingested polyphenols undergo harsh gastrointestinal digestion processes, resulting in limited bioaccessibility and low gut-epithelial permeation and bioavailability. Several lipid-based formulations have been proposed to overcome these issues. Solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) have also been highlighted as carrier systems for the oral delivery of lipophilic bioactives, including polyphenols. This paper summarizes the research on the ingredients, production methods, post-processing procedures, general characteristics, and advantages and disadvantages of SLNs and NLCs. Overall, this paper reviews the applications and perspectives of polyphenol-loaded SLNs and NLCs in foods, as well as their regulation, production, storage, and economic feasibility.

Neuron-recognizable characteristics of peptides recombined using a neuronal binding domain of botulinum neurotoxin.

Recombinant peptides were designed using the C-terminal domain (receptor binding domain, RBD) and its subdomain (peptide A2) of a heavy chain of botulinum neurotoxin A-type 1 (BoNT/A1), which can bind to the luminal domain of synaptic vesicle glycoprotein 2C (SV2C-LD). Peptide A2- or RBD-containing recombinant peptides linked to an enhanced green fluorescence protein (EGFP) were prepared by expression in Escherichia coli. A pull-down assay using SV2C-LD-covered resins showed that the recombinant peptides for CDC297 BoNT/A1, referred to EGFP-A2' and EGFP-RBD', exhibited ≥ 2.0-times stronger binding affinity to SV2C-LD than those for the wild-type BoNT/A1. Using bio-layer interferometry, an equilibrium dissociation rate constant (KD) of EGFP-RBD' to SV2C-LD was determined to be 5.45 µM, which is 33.87- and 15.67-times smaller than the KD values for EGFP and EGFP-A2', respectively. Based on confocal laser fluorescence micrometric analysis, the adsorption/absorption of EGFP-RBD' to/in differentiated PC-12 cells was 2.49- and 1.29-times faster than those of EGFP and EGFP-A2', respectively. Consequently, the recombinant peptides acquired reasonable neuron-specific binding/internalizing ability through the recruitment of RBD'. In conclusion, RBDs of BoNTs are versatile protein domains that can be used to mark neural systems and treat a range of disorders in neural systems.

Development of End-Spliced Dimeric Nanodiscs for the Improved Virucidal Activity of a Nanoperforator.

Lipid-bilayer nanodiscs (NDs) wrapped in membrane scaffold proteins (MSPs) have primarily been used to study membrane proteins of interest in a physiological environment. Recently, NDs have been employed in broader applications including drug delivery, cancer immunotherapy, bio-imaging, and therapeutic virucides. Here, we developed a method to synthesize a dimeric nanodisc, whose MSPs are circularly end-spliced, with long-term thermal stability and resistance to aggregation. The end-spliced nanodiscs (esNDs) were assembled using MSPs that were self-circularized inside the cytoplasm ofEscherichia colivia highly efficient protein trans-splicing. The esNDs demonstrated a consistent size and 4-5-fold higher stability against heat and aggregation than conventional NDs. Moreover, cysteine residues on trans-spliced circularized MSPs allowed us to modulate the formation of either monomeric nanodiscs (essNDs) or dimeric nanodiscs (esdNDs) by controlling the oxidation/reduction conditions and lipid-to-protein ratios. When the esdNDs were used to prepare an antiviral nanoperforator that induced the disruption of the viral membrane upon contact, antiviral activity was dramatically increased, suggesting that the dimerization of nanodiscs led to cooperativity between linked nanodiscs. We expect that controllable structures, long-term stability, and aggregation resistance of esNDs will aid the development of novel versatile membrane-mimetic nanomaterials with flexible designs and improved therapeutic efficacy.

Interfacial and colloidal characterization of oil-in-water emulsions stabilized by interface-tunable solid lipid nanoparticles.

We evaluated the correlation between the interfacial characteristics of solid lipid nanoparticles (SLNs) and the interfacial/colloidal stability of SLN-stabilized emulsions. Herein, the interfacial properties of SLNs, particularly the surface load (Γs) of emulsifiers, were tuned by controlling the type/concentration of emulsifier used to prepare the SLNs. Increasing the Γs decreased the contact angle at the oil-water interface, which enhanced the displacement free energy of the SLNs at the interface. Moreover, the Γs of emulsifiers bound to the surface of SLNs covering oil droplets was linearly correlated with the SLN-own Γs. The size/ζ-potential of emulsions stabilized by SLNs covered by the highest concentration of emulsifiers was unchanged for 1 month, indicating good emulsion stability. The interfacial/colloidal stability of SLN-stabilized emulsions was thus enhanced by increasing the emulsifier concentration used to produce the SLNs. This study provides baseline data for developing SLN-stabilized emulsions for the food, cosmetic, and pharmaceutical industries.

Envelope-deforming antiviral peptide derived from influenza virus M2 protein.

Molecules interfering with lipid bilayer function exhibit strong antiviral activity against a broad range of enveloped viruses, with a lower risk of resistance development than that for viral protein-targeting drugs. Amphipathic peptides are rich sources of such membrane-interacting antivirals. Here, we report that influenza viruses were effectively inactivated by M2 AH, an amphipathic peptide derived from the M2 protein of the influenza virus. Although overall hydrophobicity () of M2 AH was not related to antiviral activity, modification of the hydrophobic moment (<µH>) of M2 AH dramatically altered the antiviral activity of this peptide. M2 MH, a derivative of M2 AH with a <µH> of 0.874, showed a half maximal inhibitory concentration (IC50) of 53.3 nM against the A/PR/8/34 strain (H1N1), which is 16-times lower than that of M2 AH. The selectivity index (IC50/CC50), where CC50 is the half maximal cytotoxic concentration, was 360 for M2 MH and 81 for M2 AH. Dynamic light scattering spectroscopy and electron microscopy revealed that M2 AH-derived peptides did not disrupt liposomes but altered the shape of viruses. This result suggests that the shape of virus envelope was closely related to its activity. Thus, we propose that deforming without rupturing the membranes may achieve a high selectivity index for peptide antivirals.

Improving Flavonoid Bioaccessibility using an Edible Oil-Based Lipid Nanoparticle for Oral Delivery.

To enhance the oral bioaccessibility of flavonoids, including quercetin, naringenin, and hesperetin, we prepared an edible oil-based lipid nanoparticle (LNP) system. Flavonoid-loaded LNPs were similar to the blank LNP in physicochemical characteristics (z average <154.8 nm, polydispersity index <0.17, and ζ potential < -40.8 mV), and their entrapment efficiency was >81% at 0.3 wt % flavonoid concentration of the lipid phase. In the simulated digestion assay (mouth, stomach, and small intestine), LNPs were hydrolyzed under small intestine conditions and protected successfully incorporated flavonoids (≥94%). Moreover, the relative bioaccessibility of flavonoids was >71%, which was otherwise <15%, although flavonoids were released rapidly from LNPs into the medium. In conclusion, since the flavonoids incorporated in LNPs were preserved well during oral digestion and had improved bioaccessibility, the designed LNP system may serve as an encapsulation strategy to enhance the bioavailability of nonbioaccessible nutraceuticals in foods.

Enhancing the stability of lipid nanoparticle systems by sonication during the cooling step and controlling the liquid oil content.

Aggregation of unstable particles in water limits the application of lipid nanoparticle (LNP) systems to foods despite the capability to encapsulate lipophilic bioactive components. This study exploits a preparation process that can reduce the aggregation of LNPs. Sonication during the cooling step (postsonication) for 4, 5, or 6 min was applied to increase the covering effect of Tween 20 on the particle. Additionally, LNPs were prepared using fully hydrogenated canola oil (FHCO) blended with 0-30 wt % liquid canola oil (LCO) of the lipid phase. Surfactant surface load data indicate that the postsonication might make nonemulsifying Tween 20 diffuse from the aqueous phase to droplet surfaces, which could decrease the crystallinity index (CI) of LNPs due to the inhibition of lipid crystallization. Moreover, the LCO content in lipid matrix could decrease the CI, which could reduce the formation of hydrophobic patches on the particle surface. Therefore, the postsonication and the LCO addition in the matrix could effectively prevent aggregation among hydrophobic patches. This improved colloidal stability of LNPs was verified by the particle shape in transmission electron microscopy and the gelation test. Consequently, LNPs fabricated using 6 min postsonication and 30 wt % LCO in the lipid exhibited the greatest stability (size, 202.3 nm; CI, 57.5%; Tween 20 surface load, 10.29 mg m(-2)). This study may serve as a basis for further research that aims to develop delivery systems for functional foods.