Nanostarch-Stimulated Cell Adhesion in 3D Bioprinted Hydrogel Scaffolds for Cell Cultured Meat.

Three-dimensional (3D) bioprinting has great potential in the applications of tissue engineering, including cell culturing meat, because of its versatility and bioimitability. However, existing bio-inks used as edible scaffold materials lack high biocompatibility and mechanical strength to enable cell growth inside. Here, we added starch nanoparticles (SNPs) in a gelatin/sodium alginate (Gel/SA) hydrogel to enhance printing and supporting properties and created a microenvironment for adherent proliferation of piscine satellite cells (PSCs). We demonstrated the biocompatibility of SNPs for cells, with increasing 20.8% cell viability and 36.1% adhesion rate after 5 days of incubation. Transcriptomics analysis showed the mechanisms underlying the effects of SNPs on the adherent behavior of myoblasts. The 1% SNP group had a low gel point and viscosity for shaping with PSCs infusion and had a high cell number and myotube fusion index after cultivation. Furthermore, the formation of 3D muscle tissue with thicker myofibers was shown in the SNP-Gel/SA hydrogel by immunological staining.

Tissue-like cultured fish fillets through a synthetic food pipeline.

Tissue-like cultured meats of some livestock have successfully been established by different approaches. However, the production of a structure similar to fish fillets is still challenging. Here, we develop tissue-like cultured fish fillets by assembly of large yellow croaker muscle fibers and adipocytes with 3D-printed gel. Inhibition of Tgf-ß and Notch signals significantly promoted myogenic differentiation of piscine satellite cells (PSCs). The mixture of fish gelatin and sodium alginate combined with a p53 inhibitor and a Yap activator supported PSC viability and proliferation. Based on the texture of fish muscle tissue, a 3D scaffold was constructed by gelatin-based gel mixed with PSCs. After proliferation and differentiation, the muscle scaffold was filled with cultured piscine adipocytes. Finally, tissue-like fish fillets with 20 × 12 × 4 mm were formed, consisting of 5.67 × 107 muscles and 4.02 × 107 adipocytes. The biomanufacture of tissue-like cultured fish fillet here could be a promising technology to customize meat production with high fidelity.

Ultrasound-induced cell disintegration and its ultrastructure characterization for the valorisation of Chlorella pyrenoidosa protein.

Chlorella pyrenoidosa (CP) has great potential for feeding future demands in food, environment, energy, and pharmaceuticals. To achieve this goal, the exploitation of emerging efficient technique such as ultrasound-assisted extraction (UAE) for CP nutrient enrichment is crucial. Here, UAE is deployed for high-efficient CP protein (CPP) valorisation. Compared to conventional solvent extraction (CSE), remarkable mass transfer enhancements with 9-time protein yields and 3-time extraction rate are achieved by ultrasonic cavitation in UAE, indicating UAE can drastically shift intracellular nutrients including proteins and pigments to solvent. Cell morphology and ultrastructure show the different responses of cell wall and membrane, indicating that the cell membrane may play a role in the extraction process, based on which the extremely-low efficiency of CSE and high efficiency of UAE are highlighted. This study provides a solution for future food crisis by extracting CPP and may open a new discussion field in ultrasonic extraction.

Digestion, absorption, and transport properties of soy-fermented douchi hypoglycemic peptides VY and SFLLR under simulated gastrointestinal digestion and Caco-2 cell monolayers.

Two novel hypoglycemic peptides VY and SFLLR were identified from douchi as the major peptides responsible for the glucose uptake activity. The present work aimed to elucidate their digestion, absorption and transport properties using simulated digestion and Caco-2 cell monolayers transport models. Besides, the effects of digestion and absorption on the structure and activity were also studied. The results showed that VY was resistant to gastrointestinal tract digestion and could cross Caco-2 cell monolayers intactly via both TJs-mediated passive paracellular pathway and PepT1-mediated active route. In comparison, SFLLR was partially degraded into small fragments of SFLL, SFL, and SF by the digestive system, leading to increased glucose uptake activity. Notably, SFLLR, SFLL, and SFL were partly hydrolyzed by aminopeptidase N or dipeptidyl peptidase IV during transport, but they were transported intact. SFL was transported via both paracellular diffusion and PepT1-mediated routes, while SFLLR and SFLL were via paracellular route only.

Efficient Retention and Complexation of Exogenous Ferulic Acid in Starch: Could Controllable Bioextrusion Be the Answer?

The starch-phenolics complexes are widely fabricated as functional foods but with low phenolics retention limited by traditional liquid reaction and washing systems. In this study, ferulic acid (FA, 5%) was exogenously used in the crystalline form, and it reacted with starch in a high-solid extrusion environment, which was simultaneously controlled by thermostable α-amylase (0-252 U/g). Moderate enzymolysis (21 or 63 U/g) decreased the degree of the starch double helix and significantly increased the FA retention rate (>80%) with good melting and distribution. Although there were no significantly strong chemical bonds (with only 0.17-2.39% FA bound to starch hydrolysate), the noncovalent interactions, mainly hydrogen bonds, van der Waals forces, and electrostatic interactions, were determined by 1H NMR and molecular dynamics simulation analyses. The phased release of total FA (>50% in the stomach and ∼100% in the intestines) from bioextrudate under in vitro digestion conditions was promoted, which gives a perspective for handing large loads of FA and other phenolics based on starch carrier.

Composition and Rheological Properties of Peanut Oil Bodies from Aqueous Enzymatic Extraction.

In this study, the relationship between the composition and rheological properties of peanut oil bodies from aqueous enzymatic extraction was evaluated. Aqueous enzymatic extraction using a combination of cellulase and pectinase at a 1:1 ratio effectively destroyed the structure of the cell wall and resulted in the maximum oil body yield of 90.7%. The microstructure and interfacial membrane composition of the peanut oil bodies were observed by confocal laser scanning microscopy. The oil bodies contained three inherent proteins (oleosin, caleosin, and steroleosin) along with two adsorbed foreign proteins (arachin and lipoxygenase). Five phospholipids were detected using 31P nuclear magnetic resonance spectroscopy. Among them, phosphatidylcholine, which plays a major role in the stability of oil bodies, was the most abundant. The measured rheological properties indicated that the oil bodies were a typical elastic system. Elevated temperature and high-speed shear destroyed the binding between proteins and phospholipids, reducing the oil body stability. The findings will facilitate the commercial application of peanut oil bodies by improving the extraction rate of peanut oil bodies and clarifying their stabilization mechanism.Practical Application: This paper studies the enzymatic extraction, composition and rheological properties of peanut oil bodies. It provides a theoretical basis for the large-scale application of peanut oil bodies in the food and cosmetic industries. It is beneficial to improve the application value of peanut resources.

Effects of Pretreatment on the Yield of Peanut Oil and Protein Extracted by Aqueous Enzymatic Extraction and the Characteristics of the Emulsion.

Effects of comminution on peanut particle size and yield of peanut oil and protein were analyzed. Additionally, the emulsion properties (surface protein concentration, particle size, and ξ-potential) were compared. Moreover, different demulsification methods were used to investigate the emulsion stability. Results showed that the yield of peanut oil and protein was highest (87.23% and 82.05%, respectively) after dry comminution for 72 s. Upon wet comminution for 120 s, the yields of peanut oil and protein were 89.91% and 84.70%, respectively, which were both significantly higher than that obtained after dry comminution (p < 0.05). The surface protein concentration and ξ-potential of emulsion made by dry comminution (DCE) were 7.02 mg/m2 and 12.08 mV, respectively, and those of emulsion made by wet comminution (WCE) were 10.71 mg/m2 and 15.25 mV, respectively, which were significantly higher than that of DCE (p < 0.05). The volume average particle size of DCE was 3.41 µm, which was significantly higher than that of WCE (3.18 µm, p < 0.05). Collectively, these results indicated that WCE was more stable than DCE. Further, the demulsification rate of DCE was significantly higher than that of WCE when treated by freeze-thawing, pH, papain, and phospholipase A2 (p < 0.05). Demulsification effect of Alcalase 2.4L was the best among these five demulsification methods treated, and the demulsification rate of DCE reached 92.77%, which was slightly higher than that of WCE (92.67%), further illustrating the higher stability of WCE.

Effect of Papain on the Demulsification of Peanut Oil Body Emulsion and the Corresponding Mechanism.

This study investigated the effect of papain on the demulsification of peanut oil body emulsion extracted using an aqueous enzymatic method and the associated mechanism. The highest free oil yield using papain (92.39%) was obtained under the following conditions: an enzymatic hydrolysis temperature of 55°C, sample-to-water ratio of 1:3, enzyme concentration of 1400 U/g, and an enzymatic hydrolysis time of 3 h. Papain degraded the peanut oil body protein to small-molecular-weight peptides (≤ 14.4 kDa). Compared to the emulsion before enzymatic hydrolysis, the amino acid content in the aqueous phase was higher after enzymatic hydrolysis, the viscosity of the oil body emulsion was lower, and the particle diameter of the emulsion was significantly larger. The following demulsification mechanism was derived. Papain degrades the protein on the peanut oil body and dissolves it in water. The outer side of the oil body loses the protection of electrostatic repulsion and steric hindrance provided by the membrane protein. This causes the viscosity of the emulsion system and the molecular steric hindrance to decrease. As a result, the oil droplets gather and eventually demulsify. The results of this study provide the theoretical basis for the instability in oil body emulsions and are expected to promote the application of enzymatic demulsification in industry.