Effects of eugenol on the structure and gelling properties of myofibrillar proteins under hydroxyl radical-induced oxidative stress.

The effects of eugenol (EG; 0, 5, 20, and 50 mg/g protein) on the structure and gel properties of pork myofibrillar protein (MPs) under a hydroxyl radical-generating system were explored in this study. The results revealed that the addition of a high concentration of EG (50 mg/g protein) markedly reduced the carbonyl content and enhanced the fluorescence intensity, surface hydrophobicity, and protected the secondary structure of MPs, compared to oxidized MPs. In addition, the high concentration group noticeably increased the storage modulus (G'), gel strength, and water-holding capacity (WHC), and significantly hindered the oxidation-induced transformation of immobilized water of the MPs gel to free water and basically favored the formation of a finer and more homogeneous three-dimensional network structure, This work verified that the adding of EG could effectively improve the gel quality of oxidized MPs and more successfully delay oxidation-induced damage to muscle protein structure.

Restoration and preservation effects of mung bean antioxidant peptides on H2O2-induced WRL-68 cells via Keap1-Nrf2 pathway.

Mung bean antioxidant peptides (MBAPs) were prepared from mung bean protein hydrolysate, and four peptide sequences including Ser-Asp-Arg-Thr-Gln-Ala-Pro-His (~953 Da), Ser-His-Pro-Gly-Asp-Phe-Thr-Pro-Val (~956 Da), Ser-Asp-Arg-Trp-Phe (~710 Da), and Leu-Asp-Arg-Gln-Leu (~644 Da) were identified. The effects of MBAPs on the oxidation-induced normal human liver cell line WRL-68 were analyzed to determine the mechanism protecting the oxidation-induced injury. The results showed that the cells were subjected to certain oxidative damage by H2O2 induction, as evidenced by decreased cell number and viability, overproduction of intracellular ROS, and decreased mitochondrial membrane potential. Compared with the H2O2-induced group, the MBAP-treated oxidation-induced group exhibited significantly higher cell number and viability, and the intracellular ROS was similar to that of the control group, suggesting that MBAP scavenges excessive intracellular free radicals after acting on the oxidation-induced cells. Combined with Western blotting results, it was concluded that the MBAP-treated oxidation-induced group also significantly promoted the expression of proteins related to the kelch-like ech-related protein 1 (Keap1)/ nuclear factor e2-related factor 2 (Nrf2) signaling pathway, which resulted in an approximately 2-fold increase in antioxidant enzymes, and a decrease in malondialdehyde content of approximately 55% compared to oxidatively-induced cells, leading to the recovery of both cell morphology and viability. These results suggest that MBAPs scavenge intracellular free radicals and improve oxidative stress in hepatocytes through the expression of Keap1/Nrf2 pathway-related protein, thereby reducing oxidative attack on the liver. Therefore, MBAP is applied as a nutritional ingredient in the functional food field, and this study provides a theoretical basis for the high utilization of mung bean proteins.

Effects of Heat Treatment on the Structural and Functional Properties of Phaseolus vulgaris L. Protein.

The paper presents the effect of heat treatment at 80 °C at different times (3, 5, 7, and 9 min) on the structural and functional properties of Phaseolus vulgaris L. protein (PVP, bean protein powder). Surface and structure properties of PVP after heat treatment were analyzed using a Fourier transform infrared spectrometer (FTIR), a fluorescence spectrophotometer, a visible light spectrophotometer, a laser particle size analyzer, and other equipment. The secondary structure and surface hydrophobicity (H0) of PVP changed significantly after heat treatment: the ß-sheet content decreased from 25.32 ± 0.09% to 24.66 ± 0.09%, the random coil content increased from 23.91 ± 0.11% to 25.68 ± 0.08%, and the H0 rose by 28.96-64.99%. In addition, the functional properties of PVP after heat treatment were analyzed. After heat treatment, the emulsifying activity index (EAI) of PVP increased from 78.52 ± 2.01 m2/g to 98.21 ± 1.33 m2/g, the foaming ability (FA) improved from 87.31 ± 2.56% to 95.82 ± 2.96%, and the foam stability (FS) rose from 53.23 ± 1.72% to 58.71 ± 2.18%. Finally, the degree of hydrolysis (DH) of PVP after gastrointestinal simulated digestion in vitro was detected by the Ortho-Phthal (OPA) method. Heat treatment enhanced the DH of PVP from 62.34 ± 0.31% to 73.64 ± 0.53%. It was confirmed that heat treatment changed the structural properties of PVP and improved its foamability, emulsification, and digestibility. It provides ideas for improving PVP's potential and producing new foods with rich nutrition, multiple functions, and easy absorption.

Recent advances in exploring and exploiting soybean functional peptides-a review.

Soybeans are rich in proteins and phytochemicals such as isoflavones and phenolic compounds. It is an excellent source of peptides with numerous biological functions, including anti-inflammatory, anticancer, and antidiabetic activities. Soy bioactive peptides are small building blocks of proteins that are released after fermentation or gastrointestinal digestion as well as by food processing through enzymatic hydrolysis, often in combination with novel food processing techniques (i.e., microwave, ultrasound, and high-pressure homogenization), which are associated with numerous health benefits. Various studies have reported the potential health benefits of soybean-derived functional peptides, which have made them a great substitute for many chemical-based functional elements in foods and pharmaceutical products for a healthy lifestyle. This review provides unprecedented and up-to-date insights into the role of soybean peptides in various diseases and metabolic disorders, ranging from diabetes and hypertension to neurodegenerative disorders and viral infections with mechanisms were discussed. In addition, we discuss all the known techniques, including conventional and emerging approaches, for the prediction of active soybean peptides. Finally, real-life applications of soybean peptides as functional entities in food and pharmaceutical products are discussed.

Beta-TCP scaffolds with rationally designed macro-micro hierarchical structure improved angio/osteo-genesis capability for bone regeneration.

The design of hierarchical porous structure in scaffolds is crucial for bone defect regenerative repair. However, bioceramic materials present a challenge in precisely constructing designed micropores owing to the limitation of forming process. To investigate micropore shape influences bone regeneration in bioceramic scaffolds with macropores, hierarchical porous scaffolds with interconnective macropores (~400 µm) and two types of micropores (spherical and fibrous) were prepared using a combination of direct ink writing (DIW) and template sacrifice methods. Compared to the scaffold with spherical micropores, the scaffold with highly interconnected fibrous micropores significantly improved cell adhesion and upregulated osteogenic and angiogenetic-related gene expression in mBMSCs and HUVECs, respectively. Furthermore, in vivo implantation experiments showed that hierarchical scaffolds with fibrous micropores accelerated the bone repair process significantly. This result can be attributed to the high interconnectivity of fibrous micropores, which promotes the transportation of nutrients and waste during bone regeneration. Our work demonstrates that hierarchical porous scaffold design, especially one with a fibrous micropore structure, is a promising strategy for improving the bone regeneration performance of bioceramic scaffolds.

Concentration-dependent effect of eugenol on porcine myofibrillar protein gel formation.

The effects of different concentrations of eugenol (EG = 0, 5, 10, 20, 50, and 100 mg/g protein) on the structural properties and gelling behavior of myofibrillar proteins (MPs) were investigated. The interaction of EG and MPs decreased free thiol and amine content, and reduced tryptophan fluorescence intensity and thermal stability, but enhanced surface hydrophobicity and aggregation of MPs. Compared with the control (EG free), the MPs' gels treated with 5 and 10 mg/g of EG had a higher storage modulus, compressive strength, and less cooking loss. A high microscopic density was observed in these EG-treated gels. However, EG at 100 mg/g was detrimental to the gelling properties of the MPs. The results indicate that an EG concentration of 20 mg/g is a turning point, i.e., below 20 mg/g, EG promoted MPs gelation, but above 20 mg/g, it impeded gelation by interfering with protein network formation. The EG modification of MPs could provide a novel ingredient strategy to improve the texture of comminuted meat products.

3D printed pore morphology mediates bone marrow stem cell behaviors via RhoA/ROCK2 signaling pathway for accelerating bone regeneration.

Bone bionics and structural engineering have sparked a broad interest in optimizing artificial scaffolds for better bone regeneration. However, the mechanism behind scaffold pore morphology-regulated bone regeneration remains unclear, making the structure design of scaffolds for bone repair challenging. To address this issue, we have carefully assessed diverse cell behaviors of bone mesenchymal stem cells (BMSCs) on the ß-tricalcium phosphate (ß-TCP) scaffolds with three representative pore morphologies (i.e., cross column, diamond, and gyroid pore unit, respectively). Among the scaffolds, BMSCs on the ß-TCP scaffold with diamond pore unit (designated as D-scaffold) demonstrated enhanced cytoskeletal forces, elongated nucleus, faster cell mobility, and better osteogenic differentiation potential (for example, the alkaline phosphatase expression level in D-scaffold were 1.5-2 times higher than other groups). RNA-sequencing analysis and signaling pathway intervention revealed that Ras homolog gene family A (RhoA)/Rho-associated kinase-2 (ROCK2) has in-depth participated in the pore morphology-mediated BMSCs behaviors, indicating an important role of mechanical signaling transduction in scaffold-cell interactions. Finally, femoral condyle defect repair results showed that D-scaffold could effectively promote endogenous bone regeneration, of which the osteogenesis rate was 1.2-1.8 times higher than the other groups. Overall, this work provides insights into pore morphology-mediated bone regeneration mechanisms for developing novel bioadaptive scaffold designs.

3D printed strontium-doped calcium phosphate ceramic scaffold enhances early angiogenesis and promotes bone repair through the regulation of macrophage polarization.

The vascularization of bone repair materials is one of the key issues that urgently need to be addressed in the process of bone repair. The changes in macrophage phenotype and function play an important role in the process of vascularization, and endowing bone repair materials with immune regulatory characteristics to enhance angiogenesis is undoubtedly a new strategy to improve the effectiveness of bone repair. In order to improve the effect of tricalcium phosphate (TCP) on vascularization and bone repair, we doped strontium ions (Sr) into TCP (SrTCP) and prepared porous 3D printed SrTCP scaffolds using 3D printing technology, and studied the scaffold mediated macrophage polarization and subsequent vascularization and bone regeneration. The results of the interaction between the scaffold and macrophages showed that the SrTCP scaffold can promote the polarization of macrophages from M1 to M2 and secrete high concentrations of VEGF and PDGF-bb cytokines, which shows excellent angiogenic potential. When human umbilical vein endothelial cells (HUVECs) were co-cultured with macrophage-conditioned medium of SrTCP scaffold, HUVECs exhibited excellent early angiogenesis-promoting effects in terms of scratch healing, angiogenic gene expression, and in vitro tube formation performance. The results of in vivo bone repair experiments showed that the SrTCP scaffold formed a vascular network with high density and quantity in the bone defect area, which could increase the rate of new bone formation and advance the period of bone formation, and finally achieved a better bone repair effect. We observed a cascade effect in which Sr-doped SrTCP scaffold regulate macrophage polarization to enhance angiogenesis and promote bone repair, which may provide a new strategy for the repair of clinical bone defects.

Flavor Differences of Edible Parts of Grass Carp between Jingpo Lake and Commercial Market.

This study investigated the flavor differences among three individual parts (abdomen, back, and tail) of Jingpo Lake grass carp (JPGC) and commercial grass carp (CGC). The growing environment and fish parts influenced the volatile compounds of the fish. The highest total contents of alcohols and ethers were found in the back of JPGC (p < 0.05). The combination of an electronic tongue and electronic nose (E-nose) could effectively distinguish the flavor differences between the different parts of JPGC and CGC by principal component analysis. Both the content of total free amino acids (FAAs) and content of amino acids contributing to the sweet and fresh flavors were higher in JPGC than CGC (p < 0.05). Among the ATP-associated products, the inosine 5'-monophosphate (IMP) contents of the back and tail of JPGC were higher (p < 0.05), but the abdomen content was lower (p > 0.05) than the respective contents in the corresponding parts of CGC. Sensory evaluation shows that JPGC had a better texture, odor, and taste, compared to CGC. Correlation analysis showed that the E-nose data and FAAs were highly correlated with the content of alcohols, aldehydes, and ethers. This study showed that the flavors of the different parts of JPGC differed significantly from those of CGC.

Hydrothermal-induced changes in the gel properties of Mung bean proteins and their effect on the cooking quality of developed compound noodles.

Mung bean proteins (MBPs) are highly nutritious food ingredients, but their lack of gluten limits their use in staple foods such as noodles. In this study, MBPs were modified by hydrothermal treatment, and their gel properties and the major structural changes were analyzed at different heating temperatures (25, 65, 75, 85, 95, and 105°C), moisture contents (0, 15, 20, 25, 30, and 35%), and hydrothermal treatment times (0, 15, 30, 45, 60, and 75 min). Thereafter, the modified MBPs (MMBPs) were added to wheat noodles at substitution levels of 3, 6, and 9% to evaluate their effect on the quality of the noodles. The results showed that the hydrothermal treatment significantly improved the gel properties and water absorption capacity of the MBPs and slightly increased their disulfide bond content. When MBPs with a 25% moisture content were heated at 85°C for 60 min, their gel properties notably improved, and their structural changes were maximal. The structural changes revealed that the MBP molecule formed a macromolecular polymer because a significant protein band appeared at about 66.2 kDa. Secondary structure and microstructure analyses revealed that the MBP structure was significantly damaged and that the ß-sheet structure increased because of changes in the degree of aggregation between the protein molecules. Compared to the untreated MBPs, the MMBPs significantly improved the cooking quality and texture properties of the noodles, and the addition amount reached more than 6%, whereas that of the untreated MBPs was less than 3%. At this time, the cooking loss and the broken rate of the untreated MBPs group were about 2 times higher than that of the 6% MMBP-treated group. An analysis of changes in the water distribution, rheological properties, and microstructure revealed that intermolecular cross-linking occurred between the MMBPs and wheat dough, which improved the quality of the MMBP-treated noodles. The findings demonstrated that the MMBPs obtained by hydrothermal treatment had a positive effect on the wheat dough properties and noodle quality. These results provide a technical foundation for incorporating novel protein supplements into staple foods.

The Effectiveness of Clove Extract on Oxidization-Induced Changes of Structure and Gelation in Porcine Myofibrillar Protein.

This study aimed to investigate the structural characteristics and gelation behavior of myofibrillar proteins (MPs) with or without clove extract (CE) at different oxidation times (0, 1, 3, and 5 h). Circular dichroism spectra and Fourier transform infrared spectra showed that samples with CE addition had significantly higher α-helix content after oxidation than those without CE addition. However, prolonged oxidation (5 h) would make the effect of CE addition less pronounced. Similarly, the ultraviolet-visible (UV) spectra analysis revealed that CE controlled the oxidative stretching of the protein tertiary structure and reduced the exposure of aromatic amino acids. In addition, the particle size and turbidity values of the CE group significantly decreased after oxidation compared to the non-CE group. CE increased the gel strength by 10.05% after 5 h of oxidation, which could be observed by scanning electron microscopy (SEM) as a more homogeneous, dense, less porous, network-like gel structure. Therefore, these results showed that oxidation induced significant changes in the structure and gel properties of MPs, but the addition of CE effectively inhibited these destructive changes.

Anti-inflammatory effects of mung bean protein hydrolysate on the lipopolysaccharide- induced RAW264.7 macrophages.

The anti-inflammatory effects of mung bean protein hydrolysate (MBPH) on the lipopolysaccharide (LPS)-induced macrophages were investigated herein. MBPH was shown to affect the cell morphology, proliferation, cell cycle, cytokine levels at different culture times, and the expression level of nuclear factor-kappa B (NF-κB). The obtained results revealed that different fractions of MBPH promote cell proliferation, alter the cell cycle by decreasing the proportion of cells in the S stage and increasing the proportion of cells in the G2 stage, increase the expression of cytokines, included IL-6, IL-1ß, and TNF-α, and negatively affect the LPS-induced inflammatory cytokines. Based on the analysis of cytokine expression at different points in time, it is concluded that cytokine secretion of MBPH-treated group reaches a peak at 24 h, the result was significantly different compared to other treatment groups (P < 0.05). It can be observed that the inflammatory response induced by LPS in the MBPH-III treatment group is reduced compared with other fractions (P < 0.05). In addition, MBPH inhibits the activation of NF-κB signaling pathway by inhibiting the nuclear transcription of p65 and phosphorylation of IκBα in macrophages induced by LPS. Our results demonstrated that lower molecular weight MBPH exerted stronger anti-inflammatory effects than other molecular fractions. Thus, MBPH could be utilized as a functional food ingredient to prevent inflammation in chronic diseases.

3D-printed bioactive ceramic scaffolds with biomimetic micro/nano-HAp surfaces mediated cell fate and promoted bone augmentation of the bone-implant interface in vivo.

Calcium phosphate bio-ceramics are osteo-conductive, but it remains a challenge to promote the induction of bone augmentation and capillary formation. The surface micro/nano-topography of materials can be recognized by cells and then the cell fate are mediated. Traditional regulation methods of carving surface structures on bio-ceramics employ mineral reagents and organic additives, which might introduce impurity phases and affect the biological results. In a previous study, a facile and novel method was utilized with ultrapure water as the unique reagent for hydrothermal treatment, and a uniform hydroxyapatite (HAp) surface layer was constructed on composite ceramics (ß-TCP/CaSiO3) in situ. Further combined with 3D printing technology, biomimetic hierarchical structure scaffolds were fabricated with interconnected porous composite ceramic scaffolds as the architecture and micro/nano-rod hybrid HAp as the surface layer. The obtained HAp surface layer favoured cell adhesion, alleviated the cytotoxicity of precursor scaffolds, and upregulated the cellular differentiation of mBMSCs and gene expression of HUVECs in vitro. In vivo studies showed that capillary formation, bone augmentation and new bone matrix formation were upregulated after the HAp surface layer was obtained, and the results confirmed that the fabricated biomimetic hierarchical structure scaffold could be an effective candidate for bone regeneration.

Hierarchically porous calcium-silicon nanosphere-enabled co-delivery of microRNA-210 and simvastatin for bone regeneration.

The regenerative repair of large bone defects is a major problem in orthopedics and clinical medicine. The key problem is the lack of ability of existing bone graft materials to promote osteogenesis and angiogenesis. Previous studies have shown that the osteogenic or angiogenic abilities of these materials could be significantly improved by adding miRNA or small-molecule drugs to bone graft materials; however, the synergistic effect arising from this combination is not clear. Therefore, we proposed to construct a dual drug delivery system that could simultaneously achieve the co-encapsulation and co-delivery of miRNA and small-molecule drugs to explore the effect of a dual drug delivery system on bone repair. In this study, we constructed dual-sized pore structure calcium-silicon nanospheres (DPNPs) and achieved the co-encapsulation of miR-210, angiogenic gene drugs, and simvastatin (Siv), a small-molecule osteogenic drug, through metal-ion coordination and physical adsorption. In vitro and in vivo osteogenic and angiogenic experiments showed that the dual drug delivery system (Siv/DPNP/miR-210) exhibited better properties than those of the individual unloaded and single drug-loaded systems and could significantly accelerate the process of bone repair, which provides a novel strategy for the regeneration and repair of bone defects.

Cellulose Nanofiber/Carbon Nanotube Dual Network-Enabled Humidity Sensor with High Sensitivity and Durability.

Humidity sensors have been widely used for humidity monitoring in industrial fields, while the unsatisfactory flexibility, time consumption, and expensive integration process of conventional inorganic sensors significantly limit their application in wearable electronics. Using paper-based humidity sensors is considered a feasible method to overcome these drawbacks because of their good flexibility and roll-to-roll manufacturability, while they still face problems such as poor durability and low sensitivity. In this study, we report a high-performance paper-based humidity sensor based on a rationally designed bilayered structure consisting of a nanoporous cellulose nanofiber/carbon nanotube (CNF/CNT) sensitive layer and a microporous paper substrate. The vast number of hydrophilic hydroxyl groups on the surface of CNF and paper fibers enables fast water molecule exchange between the humidity-sensitive material and the external environment via hydrogen bonding, endowing the paper-based sensor with an excellent humidity responsive property. The obtained sensor displays a maximum response value of 65.0% (ΔI/I0) at 95% relative humidity. Furthermore, the mechanical interlocking structure formed between the CNF/CNT layer and the paper layer provides the sensor with strong interlayer adhesion. Benefiting from the unique structure, the sensor also exhibits outstanding bending (with a maximum curvature of 22.2 cm-1) and folding durability (up to 50 times). Finally, as a proof of concept, a simple humidity-measuring device is assembled, which demonstrates an excellent responsive property toward human breath and the change of air humidity, indicating a great potential of our paper-based humidity sensor toward practical applications.

Mung Bean Protein Hydrolysate Modulates the Immune Response Through NF-κB Pathway in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages.

The objective of this study was to evaluate the immunomodulatory activity of mung bean protein hydrolysate (MBPH) in lipopolysaccharide (LPS)-induced RAW 264.7 cells and discuss the possible immune regulatory mechanism. MBPH was prepared by alcalase, trypsin, neutrase, and flavourzyme. The 3-h alcalase-hydrolyzed hydrolysate with a molecular weight less than 1,450 Da was selected for the immunological tests. Results showed that MBPH possessed strong suppressing activity to proinflammatory mediators in a dose-dependent manner. Compared to the LPS alone group, MBPH (200 µg/mL) significantly reduced nitric oxide (NO), inducible nitric oxide synthase, interleukin (IL)-6, and IL-1ß secretion levels by 52.6%, 53.2%, 48.4%, and 49.7%, respectively, in LPS-induced macrophages. It also enhanced IL-10 secretion from 789 to 3,678 pg/mL. MBPH blocked nuclear factor-kappa B (NF-κB) translocation in LPS-induced macrophages through the prevention of IκBα phosphorylation, and this process further prevented p65 translocation into the nucleus. A possible mechanism of MBPH is that it regulated the expression of inflammatory factors via the NF-κB pathway, thus inhibiting inflammatory reactions. The results suggested that MBPH is of application potential in the development of immunomodulatory functional food to ameliorate immunosuppression.

3D-Plotted Beta-Tricalcium Phosphate Scaffolds with Smaller Pore Sizes Improve In Vivo Bone Regeneration and Biomechanical Properties in a Critical-Sized Calvarial Defect Rat Model.

Due to the difficulty in fabricating bioceramic scaffolds with smaller pore sizes by the current 3D printing technique, the effect of smaller pore sizes (below 400 µm) of 3D printed bioceramic scaffolds on the bone regeneration and biomechanical behavior is never studied. Herein beta-tricalcium phosphate (ß-TCP) scaffolds with interconnected smaller pores of three different sizes (100, 250, and 400 µm) are fabricated by 3D plotting. The resultant scaffolds are then implanted into rat critical-sized calvarial defects without any seeded cells. A custom-designed device is developed to investigate the biomechanical properties of the scaffolds after surgical implantation for 4, 8, and 12 weeks. The scaffolds with the 100 µm pore size are found to present the highest maximum load and stiffness, comparable to those of the autogenous bone, after being implanted for 12 weeks. Micro-computed tomography (micro-CT) and histological analysis further indicate that the scaffolds with the 100 µm pore size achieve the highest percentage of new bone ingrowth, which correlates to their best in vivo biomechanical properties. This study demonstrates that tailoring the pore size of ß-TCP scaffolds to a smaller range by 3D-plotting can be a facile and efficient approach to enhanced bone regeneration and biomechanical behaviors in bone repair.

The effectiveness of clove extracts in the inhibition of hydroxyl radical oxidation-induced structural and rheological changes in porcine myofibrillar protein.

Oxidation is a major cause of protein quality deterioration during the storage and processing of food. This study investigated the effects of clove extract (CE) on structural and rheological changes in porcine longissimus myofibrillar proteins (MP) and the effects of oxidizing radicals produced by a Fenton reaction system (FRS). Increased oxidation time was accompanied by increased carbonyl content, reduced Ca-ATPase activity, decreased enthalpy of denaturation, decreased thermal transition temperatures (P<0.05), and increased protein susceptibility to thermal aggregation. The addition of CE significantly inhibited carbonyl formation (P<0.05), enhanced solubility and thermal stability, and improved the gel formation ability (storage modulus, loss modulus) of MP. The protective effect of CE on protein denaturation was demonstrated by its efficacy in maintaining Ca-ATPase activity and decreasing the degree of protein aggregation. Overall, the hydroxyl radical-induced loss of the structural and functional properties of MP was significantly reduced by the presence of CE.