Fabrication of high internal phase Pickering emulsions with calcium-crosslinked whey protein nanoparticles for ß-carotene stabilization and delivery.

Whey protein isolate (WPI) nanoparticles were fabricated with Ca2+ induced cross-linking and used as an effective particle stabilizer for high internal phase Pickering emulsion (HIPPE) formulation aiming to improve the chemical stability and bioaccessibility of ß-carotene (BC). Ca2+ concentration dominated the characteristics of WPI nanoparticles. Spherically shaped and homogeneously dispersed WPI nanoparticles with a Z-average diameter of approximately 150.0 nm were obtained with 5.0 mM Ca2+ concentration. No cytotoxicity was observed for WPI nanoparticles even at 10.0 mg mL-1 concentration. HIPPE (oil fraction 80.0%, w/w) can be successfully prepared with WPI nanoparticles at a concentration as low as 0.2% (w/w) and was stable for at least 2 months at room temperature. A higher WPI nanoparticle concentration resulted in more solid-like HIPPEs. BC exhibited appreciably higher retention in HIPPEs than in bulk oil during 30 days of storage at 50 °C. Moreover, BC bioaccessibility was appreciably improved with the HIPPE delivery system. Both the chemical stability and bioaccessibility of BC increased with the increase of WPI nanoparticle concentrations from 0.2 to 1.0% (w/w). The results obtained in this study may facilitate the fabrication of edible and biocompatible protein-based nanoparticle stabilizers for HIPPE formulation with more innovative and tailored functionalities.

α-Lactalbumin and chitosan core-shell nanoparticles: resveratrol loading, protection, and antioxidant activity.

Resveratrol (RES)-loaded protein-polysaccharide nanoparticles were fabricated through simple electrostatic interactions with oppositely charged α-lactalbumin (ALA) and chitosan (CHI) with a mass ratio of 5 : 1 without the addition of NaCl at pH 6.5. The Z-average diameter and zeta-potential values of RES-ALA-CHI nanoparticles were 211.0 nm and 13.23 mV, respectively. Both TEM and AFM graphs confirmed that RES-ALA-CHI nanoparticles had a spherical shape, and were dispersed homogeneously at the nanoscale. The encapsulation efficiency (EE) and loading amount (LA) of RES in RES-ALA-CHI nanoparticles were 58.86% and 196.2 µg mg-1, respectively, in the presence of 400 µg mL-1 RES. XRD results confirmed that RES was in amorphous form in ALA-CHI nanoparticles. The interaction between RES and ALA-CHI nanoparticles was mainly driven by hydrophobic interaction and hydrogen bonding. Compared to RES (free), the UV light and heat stability, in vitro bioaccessibility, and antioxidant activity of RES in RES-ALA-CHI nanoparticles were pronouncedly enhanced. The information provided in this study should be of interest to the food industry to fabricate robust nanoscale delivery systems with ALA-CHI nanoparticles for RES and other hydrophobic bioactive compounds.

Rapid and Accurate Determination of Nanopore Ionic Current Using a Steric Exclusion Model.

Nanopore sensing has emerged as a versatile approach to detection and identification of biomolecules. Presently, researchers rely on experience and intuition for choosing or modifying the nanopores to detect a target analyte. The field would greatly benefit from a computational method that could relate the atomic-scale geometry of the nanopores and analytes to the blockade nanopore currents they produce. Existing computational methods are either computationally too expensive to be used routinely in experimental laboratories or not sensitive enough to account for the atomic structure of the pore and the analytes. Here, we demonstrate a robust and inexpensive computational approach-the steric exclusion model (SEM) of nanopore conductance-that is orders of magnitude more efficient than all-atom MD and yet is sensitive enough to account for the atomic structure of the nanopore and the analyte. The method combines the computational efficiency of a finite element solver with the atomic precision of a nanopore conductance map to yield unprecedented speed and accuracy of ionic current prediction. We validate our SEM approach through comparison with the current blockades computed using the all-atom molecular dynamics method for a range of proteins confined to a solid-state nanopore, biological channels embedded in a lipid bilayer membranes, and blockade currents produced by DNA homopolymers in MspA. We illustrate potential applications of SEM by computing blockade currents produced by nucleosome proteins in a solid-state nanopore, individual amino acids in MspA, and by testing the effect of point mutations on amino acid distinguishability. We expect our SEM approach to become an integral part of future development of the nanopore sensing field.

Enhanced pH and thermal stability, solubility and antioxidant activity of resveratrol by nanocomplexation with α-lactalbumin.

The health-promoting benefits of resveratrol (RES) have attracted significant attention. Poor water solubility and chemical stability, however, hindered its application. In this study, α-lactalbumin (ALA)-RES nanoparticles were prepared by a simple nanocomplexation protocol. The particle sizes of ALA and the ALA-RES complex were 87.8, and 95.3 nm, respectively. AFM confirmed that both nanoparticles were spherical. XRD results confirmed that RES in ALA nanoparticles was amorphous. Fluorescence spectroscopy and FTIR showed that the ALA-RES nanocomplex was formed mainly by hydrophobic interactions. The water solubility increased by 32 times, compared to that of free RES. ALA nanocomplexation also appreciably improved the chemical stability of RES under all storage conditions, especially at pH 8.0 and high temperature. The ALA-RES nanocomplex showed significantly higher in vitro antioxidant activity than free RES. The results showed that the simple ALA-RES nanocomplex has the potential to be used as an effective antioxidant. The information obtained may enable the expanded application of ALA as an effective nanoscale carrier for delivering RES or other lipophilic nutraceuticals in the functional food, biomedical, and pharmaceutical products.

Fabrication of Resveratrol-Loaded Whey Protein-Dextran Colloidal Complex for the Stabilization and Delivery of ß-Carotene Emulsions.

The effects of resveratrol (RES)-loaded whey protein isolate (WPI)-dextran nanocomplex on the physicochemical stability of ß-carotene (BC) emulsions were evaluated. WPI-dextran was prepared by Maillard-based glycation and confirmed with gel electrophoresis and OPA assay. WPI-RES and WPI-dextran-RES nanoparticles were prepared with a simple nanocomplexation protocol. Fluorescence spectra indicated that hydrophobic interaction was the main driving force for the WPI-dextran-RES nanocomplex. Spherical and uniformly dispersed structures as well as nanoscale Z-average size (<100 nm) were confirmed for WPI-RES and WPI-dextran-RES nanocomplex with DLS and TEM. The Z-average diameter of emulsions with WPI-dextran conjugate was remarkably lower than that with WPI. Environmental stress (ionic strength, heat, and pH) and storage stability were pronouncedly improved. The chemical stability of BC with WPI-dextran-RES and WPI-RES was also remarkably enhanced when exposed to UV light and thermal treatment. The advantages of the WPI-dextran-RES colloidal complex may provide a better alternative to effectively protect and deliver hydrophobic nutraceuticals.

Oxidative stability and in vitro digestion of menhaden oil emulsions with whey protein: Effects of EGCG conjugation and interfacial cross-linking.

The goal of this study was to improve the chemical stability of menhaden oil and control the lipolysis in emulsions with whey protein during in vitro digestion through EGCG conjugation and genipin-mediated interfacial cross-linking (CL). WPI-EGCG conjugate was successfully synthesized, confirmed by SDS-PAGE, ESI-MS, and phenolic group quantifications (125.3 mg/g), and characterized with far UV CD and ATR-FTIR. Emulsion particle diameter with WPI-EGCG is lower than with WPI. Compared to the native emulsion, WPI CL increased particle diameter and physical stability. Higher oxidative stability was observed for emulsions stabilized with WPI-EGCG conjugate than that with interfacial cross-linking due to the great antioxidant activity. Whereas, WPI CL is more effective than WPI-EGCG conjugate in hindering the rate and extent of lipolysis. The combination of EGCG conjugation and interfacial CL showed both the highest protection of menhaden oil against degradation and highest inhibition on the rate and extent of lipolysis of menhaden oil.

Improved chemical stability and cellular antioxidant activity of resveratrol in zein nanoparticle with bovine serum albumin-caffeic acid conjugate.

In this study, bovine serum albumin (BSA)-caffeic acid (CA) conjugate was prepared with free radical-induced grafting method. The CA to BSA ratio of the conjugate was 115.7 mg/g. In vitro antioxidant activity assays suggested that BSA-CA conjugates had stronger antioxidant activity than BSA. Resveratrol-loaded zein encapsulated with BSA and BSA-CA conjugate core-shell nanoparticles were prepared with antisolvent method. Particle sizes were 206.3 nm, and 217.2 nm for BSA and BSA-CA, respectively. The encapsulation efficiencies (EEs) were 85.3% and 86.5% for zein-BSA and zein-BSA-CA nanoparticles, respectively. SEM results indicated that both nanoparticles were spherical with mean diameter approximately 200 nm and smooth surfaces. Both thermal and UV light stability of resveratrol was significantly improved after nanoencapsulation. BSA-CA conjugate showed remarkably greater protection than BSA against resveratrol degradation. Cellular antioxidant activity (CAA) study confirmed that resveratrol in both zein-BSA and zein-BSA-CA nanoparticles had significant higher antioxidant activities than resveratrol alone.

Development of ß-Carotene-Loaded Organogel-Based Nanoemulsion with Improved In Vitro and In Vivo Bioaccessibility.

ß-Carotene (BC), a naturally occurring lipophilic carotenoid, is beneficial for human health. However, its water solubility and bioavailability are low. In this study, organogel-based nanoemulsion was successfully prepared to improve the loading amount, solubility, and bioavailability of BC. Corn oil was selected as the oil phase for the organogel as a result of the greatest release amount of BC. Tween 20 was optimized as the emulsifier based on the highest extent of lipolysis and BC bioaccessibility. The nanoemulsion was a better alternative than the organogel according to both the extent of lipolysis and BC bioaccessibility. Cellular uptake of BC was significantly improved through organogel-based nanoemulsion compared to BC suspension. Caveolae-/lipid-raft-mediated route was the main endocytosis pathway. Pharmacokinetic results confirmed that the in vivo bioavailability of BC in nanoemulsion was 11.5-fold higher than that of BC oil. The information obtained suggested that organogel-based nanoemulsion may be an effective encapsulation system for delivery of insoluble and indigestible bioactive compounds.