Highly Adhesive Amyloid-Polyphenol Hydrogels for Cell Scaffolding.

Rationally designing microstructures of soft hydrogels for specific biological functionalization is a challenge in tissue engineering applications. A novel and affordable soft hydrogel scaffold is constructed here by incorporating polyphenol modules with lysozyme amyloid fibrils (Lys AFs) via non-covalent self-assembly. Embedded polyphenols not only trigger hydrogel formation but also determine gel behavior by regulating the polyphenol gallol density and complex ratio. The feasibility of using a polyphenol-Lys AF hydrogel as a biocompatible cell scaffold, which is conducive to cell proliferation and spreading, is also shown. Notably, introducing polyphenols imparts the corresponding hydrogels a superior cell bioadhesive efficiency without further biofunctional decoration and thus may be successfully employed in both healthy and cancer cell lines. Confocal laser scanning microscopy also reveals that the highly expressed integrin-mediated focal adhesions form due to stimulation of the polyphenol-AF composite hydrogel, direct cell adhesion, proliferation, and spreading. Overall, this work constitutes a significant step forward in creating highly adhesive tissue culture platforms for in vitro culture of different cell types and may greatly expand prospects for future biomaterial design and development.

Fabrication and characterization of Pickering emulsions stabilized by desalted duck egg white nanogels and sodium alginate.

BACKGROUND: The waste of salted egg white resources has always been a serious problem in the food industry. In this current study, we report on a kind of Pickering emulsion system, which was stabilized by duck egg white nanogels (DEWNs) and sodium alginate (SA), followed by which this system was crosslinked by calcium carbonate (CaCO3 ) via controlling the gluconolactone (GDL) concentrations, aiming to open up a promising route for making full use of these protein resources. RESULTS: The droplet size of the emulsion exhibited a reduction with an increase in SA concentrations, indicating that higher negative charges and steric hindrance was useful for a stable emulsion system. Meanwhile, the result of rheology measurement showed that storage modulus (G') values were higher than loss modulus (G″) values of the samples at higher GDL concentration, revealing the formation of elastic gel-like networks in the system, which was fabricated by SA and Ca2+ released by the CaCO3 particles. The gel-like network structure in the continuous phase improved both the freeze-thaw and thermal stability of the obtained Pickering emulsion system. Encouragingly, the Pickering high internal phase emulsions (HIPEs, φ = 0.75) stabilized by DEWN/SA3 -GDL3 were prepared, which could be stored at 4 °C for at least 30 days without oiling-off and creaming. CONCLUSION: These findings not only develop a green ultra-stable Pickering emulsion system but also extend the potential commercial applications of duck egg white proteins in the food, cosmetics, and pharmaceutical industries. © 2021 Society of Chemical Industry.

Enzymatic hydrolysis re-endows desalted duck egg white nanogel with outstanding foaming properties.

Heat-induced gel-assisted desalination could efficiently and inexpensively remove salt from salted egg whites. However, it was at the expense of the excellent foaming properties of egg whites, caused by the denaturation and aggregation of proteins during heating treatment. Hence, in this current work, the enzymatic treatment was used to re-endow duck egg white nanogels (DEWN) with outstanding foaming properties. We found that low levels of hydrolysis (DH = 2.27 %) could dramatically improve the foaming capability (FC), reaching >200 %, which also enhanced the foaming stability (FS). As the hydrolysis time extended, the adsorption and diffusion rate of the supernatant on the interface increased and performed high elasticity. The dilatational rheology and Lissajous plots were explored to investigate the nonlinear dilatational rheological behaviors of the air/water interface stabilized by the hydrolysed samples. Finally, we evaluated the effect of pH on foaming properties and found that the FC could exceed 250 %, and the FS was close to 80 % at pH = 5. These encouraging results showed that simple enzymatic treatment could revive nanogels from their dissatisfied foaming properties. In this work, gel-assisted desalination combined with enzyme treatment significantly promotes the high-quality and high-value utilization of salted egg white.

Improvement of O/W emulsion performance by adjusting the interaction between gelatin and bacterial cellulose nanofibrils.

This study was designed to improve the stability of medium internal phase emulsion by adjusting the electrostatic interaction between gelatin (GLT) and TEMPO-oxidized bacterial cellulose nanofibrils (TOBC). The influences of polysaccharide-protein ratio (1:10, 1:5, and 1:2.5) and pH (3.0, 4.7, 7.0, and 11.0) on the emulsion properties were investigated. The droplet size of TOBC/GLT-stabilized emulsion was increased with the TOBC proportion increasing at pH 3.0-11.0. Additionally, emulsion had a larger droplet size at pH 4.7 (the electrical equivalence point pH of mixtures). However, the addition of TOBC significantly improved the emulsion stability. The emulsions prepared with TOBC/GLT mixtures (mixing ratio of 1:2.5) at pH 3.0-7.0 were stable without creaming during the storage. It was because the formation of nanofibrils network impeded the droplet mobility, and the emulsion viscosity and viscoelastic modulus were increased with the addition of TOBC. These findings were meaningful to modulate the physical properties of emulsions.

Tuning of Molecular Interactions between Zein and Tannic Acid to Modify Sunflower Sporopollenin Exine Capsules: Enhanced Stability and Targeted Delivery of Bioactive Macromolecules.

There are multiple obstacles for the storage and digestion of orally administered bioactive macromolecules. This study developed a low-cost and sustained-release delivery system (sporopollenin exine capsules with zein/tannic acid modification) of proteins with excellent storage stability, and at the same time provided insights into the sustained-release mechanism through exploring the interaction between zein and tannic acid (TA). ß-Galactosidase (ß-Gal) was utilized as a model protein and loaded into sporopollenin exine capsules (SECs), which were then coated with the zein/TA system. Under the optimized zein/TA conditions, the zein/TA system showed better performance than the zein alone system in the sustained release of ß-Gal, with the residual activity of about 70.26% after 24 h of simulated digestion. Evaluation of the storage stability demonstrated a ß-Gal residual activity of nearly 90% for 28 days at 25 °C. Additionally, FTIR analysis demonstrated that the stability of the zein/TA system depends on both hydrogen bonding and certain covalent bonding through the Schiff-base reaction, and the sustained release is regulated by the bonding strength.

Coalescence behavior of eco-friendly Pickering-MIPES and HIPEs stabilized by using bacterial cellulose nanofibrils.

O/W Pickering emulsions containing oil phase with different volume fractions (50-75 v%) were facilely prepared by using bacterial cellulose nanofibrils (BCNFs) alone. The effect of oil phase volume, storage time on the surface coverage, and coalescence rate of the Pickering-MIPEs and HIPEs (medium internal phase emulsions/high internal phase emulsions) were investigated. The Pickering-MIPEs/HIPEs exhibited excellent physical stability and low coalescence rate with droplet size varying from 32 to 91 µm. The increasing of particle contents could obviously decrease the droplet size and enhance the stability of the emulsions by strengthening the network structure and increasing the steric hindrance. The result of rheology analysis confirmed the formation of a three-dimensional network, endowing the exceptional stability of the emulsions. The emulsions revealed superb stability against a wide temperature (4-50 °C) range and salt condition (0-100 mM). This novel eco-friendly Pickering-MIPEs and HIPEs would provide great opportunities for their effective utilization in green-labelled food industry.

Designable Carboxymethylpachymaran/Metal Ion Architecture on Sunflower Sporopollenin Exine Capsules as Delivery Vehicles for Bioactive Macromolecules.

There are multiple obstacles in the gastrointestinal tract (GIT) for oral administration of bioactive macromolecules. Here, we engineered an oral delivery vehicle (sporopollenin exine capsules with carboxymethylpachymaran (CMP)/metal ion modification) with targeted release based on food-grade ingredients and processing operations. Then, the interaction and binding mechanisms between CMP and metal ions in the vehicle were investigated. By using ß-galactosidase (ß-Gal) as a model protein, the systems were characterized for the surface morphology and monitored by the in vitro release profile of ß-Gal. Notably, the CMP/metal ion systems not only markedly decreased the CMP dosage but also achieved a valid long-term release compared with the previously reported CMP system. Among all the systems, the CMP/3% AlCl3 system showed the best ability to control the release with the maximum residual activity of ß-Gal at nearly 72% after 24 h of treatment. Subsequently, the interaction mechanism between CMP and metal ions within the system was characterized by the perspectives of microstructure, rheological properties, and spectroscopy characteristics. The results indicated that the low pH conditions are conducive to the further cross-linking of CMP and metal ions, resulting in a high gel strength and thus a dense structure, which can impact the controlled release of ß-Gal in the GIT. Overall, the system may be utilized in the administration of medical and functional foods, specifically for the delivery of bioactive proteins via the oral route.

Impact of plant extract on the gastrointestinal fate of nutraceutical-loaded nanoemulsions: phytic acid inhibits lipid digestion but enhances curcumin bioaccessibility.

The impact of phytic acid on lipid digestion and curcumin bioaccessibility in oil-in-water nanoemulsions was investigated using a simulated gastrointestinal tract (GIT). The size, charge, and structural organization of the colloidal particles in the system were measured as the curcumin-loaded emulsions (7 mg curcumin per g lipid) were passed through simulated mouth (pH 6.8, 2 min), stomach (pH 2.5, 2 hours), and small intestine (pH 7.0, 2 hours) stages. After the small intestine stage, the level of free fatty acids (FFAs) generated and the bioaccessibility of curcumin were measured. The total amount of FFAs released significantly decreased with increasing phytic acid level, from 105.7 ± 5.9% (control) to 78.4 ± 6.4% (0.5% phytic acid). Conversely, curcumin bioaccessibility significantly increased from 39.4 ± 3.5% (control) to 74.7 ± 2.6% (0.5% phytic acid). The inverse relationship between lipolysis and curcumin bioaccessibility was ascribed to the impact of phytic acid on droplet flocculation and the level of free calcium ions present, which affected the production of mixed micelles capable of solubilizing the nutraceutical. The knowledge obtained here might prove beneficial for the employment of phytic acid as a multifunctional ingredient that inhibits lipid digestion while boosting nutraceutical bioavailability.

Carboxymethylpachymaran entrapped plant-based hollow microcapsules for delivery and stabilization of ß-galactosidase.

ß-Galactosidase (ß-Gal) as a dietary supplement can alleviate symptoms of lactose intolerance. However, ß-Gal is deactivated due to the highly acidic conditions and proteases in the digestive tract. In this work, ß-Gal was encapsulated into L. clavatum sporopollenin exine capsules (SECs) to fabricate an oral-controlled release system and increase the stability of ß-Gal in the digestive tract. The SEC extraction process was optimized. A 3-hour vacuum loading was determined as the optimal loading time. Five different initial ratios of SECs : ß-Gal were optimized with the maximum enzyme retention rate reaching 79.40 ± 1.96%. Furthermore, ß-Gal-loaded SECs entrapped in carboxymethylpachymaran (CMP) could control the release of ß-Gal under simulated gastrointestinal conditions (SGC). The optimal enzyme retention rate reached 65.33 ± 1.46% within 24 h under SGC. Collectively, these results indicated that the entrapped SECs could be used as an effective oral delivery vehicle of ß-Gal to improve its performance as a dietary supplement in the digestion of lactose.

Phosphoprotein/chitosan electrospun nanofibrous scaffold for biomineralization.

In this study, negatively charged phosvitin (PV) and positively charged chitosan (CS) were alternately deposited on negatively charged cellulose mats via layer-by-layer (LBL) self-assembly technique. Morphologies of the LBL films coating mats were observed by scanning electron microscope (SEM). Afterwards, in vitro biomimetic mineralization was carried out through incubation of the fibrous mats in a simulated body fluid (SBF) solution for different time. Scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD) were used to characterize the morphology and structure of the deposited mineral phase on the scaffolds. In addition, the cell culture experiment demonstrated that the scaffolds with the LBL structured films were of good cell compatibility for MC3T3-E1 cells. Moreover, the cell proliferation was affected by the number of deposition layers and the composition of outer-most layer. Confocal laser scanning microscopy (CLSM) and SEM imaging revealed a good performance of cell adhesion and spreading of MC3T3-E1 cells on the surface of biocomposite scaffold. So CS/PV nanofibrous mats were satisfactory for the composite to be used in bioapplications.

Construction of pH-sensitive lysozyme/pectin nanogel for tumor methotrexate delivery.

Novel nano-particles were developed from lysozyme-pectin through self-assembly, and the nanogels could be used as a carrier for the antitumor agent, methotrexate (MTX). The nanogels exhibited spherical with diameters about 109 ± 2 nm and narrow particle size distribution, as well as negative surface charge. Furthermore, the particle size and morphology of the nanogels hardly changed with the incorporation of MTX. The loading capacity of MTX in nanogels could reach 17.58 ± 0.85%. MTX-loaded nanogels were pH-dependent, accelerated release of MTX at a decreasing pH from 7.4 to 5.3. The MTT assay indicated that encapsulated MTX exhibited higher anticancer activity than free MTX. Meanwhile, MTX-loaded nanogels could be effectively endocytosed by HepG2 cells, resulting in enhanced cancer-cell apoptosis comparing to free MTX. It indicated that the nanogels had good biocompatibility and low toxicity. The obtained nanogels had great potential in the development of a new nanocarrier for anti-cancer drug delivery.

Green-step assembly of low density lipoprotein/sodium carboxymethyl cellulose nanogels for facile loading and pH-dependent release of doxorubicin.

In this study, a simple and green approach was developed to produce a novel nanogel via self-assembly of low density lipoproteins (LDL) and sodium carboxymethyl cellulose (CMC), to efficiently deliver doxorubicin (DOX) to cancer cells. Under optimal conditions, the stable nanogels were of spherical shape with an average diameter of about 90 nm, PDI<0.3 and a zeta potential -35 mV. Furthermore, the cationic anticancer drug, doxorubicin (DOX) was effectively encapsulated into LDL/CMC nanogels with an exceptionally high encapsulation efficiency of ∼ 98%. The release of DOX from DOX-LDL/CMC nanogels was pH-dependent, and DOX was released at a quicker rate at pH 6.2 than at pH 7.4. Importantly, the DOX-LDL/CMC nanogels were shown to effectively kill cancer cells in vitro. The IC50 of the DOX-LDL/CMC nanogels in HeLa and HepG2 cells was approximately 2.45 and 1.72 times higher than that of free DOX. The slightly reduced antitumor efficacy was primarily due to the less cellular uptake of the DOX-LDL/CMC nanogels, which was confirmed by confocal laser scanning microscope (CLSM) and flow cytometry analysis. The high DOX payload and pH-dependent drug release rendered LDL/CMC nanogels as an efficient carrier for doxorubicin and possibly be used for other cationic drugs in different biomedical applications.