Effect of Porosity on the Colonization of Digital Light-Processed 3D Hydrogel Constructs toward the Development of a Functional Intestinal Model.

With the rapid increase of the number of patients with gastrointestinal diseases in modern society, the need for the development of physiologically relevant in vitro intestinal models is key to improve the understanding of intestinal dysfunctions. This involves the development of a scaffold material exhibiting physiological stiffness and anatomical mimicry of the intestinal architecture. The current work focuses on evaluating the scaffold micromorphology of gelatin-methacryloyl-aminoethyl-methacrylate-based nonporous and porous intestinal 3D, intestine-like constructs, fabricated via digital light processing, on the cellular response. To this end, Caco-2 intestinal cells were utilized in combination with the constructs. Both porous and nonporous constructs promoted cell growth and differentiation toward enterocyte-like cells (VIL1, ALPI, SI, and OCLD expression showed via qPCR, ZO-1 via immunostaining). The porous constructs outperformed the nonporous ones regarding cell seeding efficiency and growth rate, confirmed by MTS assay, live/dead staining, and TEER measurements, due to the presence of surface roughness.

Visualisation of microalgal lipid bodies through electron microscopy.

In this study, transmission electron microscopy (TEM) and cryo-scanning electron microscopy (cryo-SEM) were evaluated for their ability to detect lipid bodies in microalgae. To do so, Phaeodactylum tricornutum and Nannochloropsis oculata cells were harvested in both the mid-exponential and early stationary growth phase. Two different cryo-SEM cutting methods were compared: cryo-planing and freeze-fracturing. The results showed that, despite the longer preparation time, TEM visualisation preceded by cryo-immobilisation allows a clear detection of lipid bodies and is preferable to cryo-SEM. Using freeze-fracturing, lipid bodies were rarely detected. This was only feasible if crystalline layers in the internal structure, most likely related to sterol esters or di-saturated triacylglycerols, were revealed. Furthermore, lipid bodies could not be detected using cryo-planing. Cryo-SEM is also not the preferred technique to recognise other organelles besides lipid bodies, yet it did reveal chloroplasts in both species and filament-containing organelles in cryo-planed Nannochloropsis oculata samples.

Multiscale analysis of triglycerides using X-ray scattering: implementing a shape-dependent model for CNP characterization.

In the last decade, research has focused on examining the fundamental interactions occurring in triglycerides, aiming to comprehend the self-assembly of crystalline nanoplatelets (CNPs) and their role in forming larger hierarchical structures essential for fat functionality. Microscopy research on CNPs frequently requires disruptive preparatory techniques, such as deoiling and sonication, to achieve quantitative outcomes. Conversely, X-ray scattering has proven to be an advantageous method for studying triglycerides, as little sample is needed to quantify the system's hierarchical structures. Specifically, ultra-small-angle X-ray scattering (USAXS) has emerged as a fitting technique for studying CNPs, owing to its length scale range falling between 25 nm and 3.49 µm. In this study, we characterized four different 30% fat dilutions of stearic acid-based fats in triolein, with various purities and preparation protocols. Samples were characterized by combining diverse microscopy techniques (cryo-SEM, TEM, polarized light and phase contrast microscopy) with synchrotron-radiation X-ray scattering (WAXS, SAXS, and USAXS). A shape-dependent model for the interpretation of USAXS data is proposed, overcoming some of the drawbacks linked to previously utilized models. CNPs are modeled as polydisperse parallelepipeds, and the aggregates are characterized by fractal dimensionality. This model offers novel insights into CNP cross-section, as well as aggregation. In the long run, we hope that the model will increase our understanding of CNP conformation and interactions, helping us design new fat systems on the mesoscale.

Recent progress in oil-in-water-in-oil (O/W/O) double emulsions.

Oil-in-water-in-oil (O/W/O) double emulsions are recognized as an advanced design route for oil structuring that shows promising applications in the pharmaceutical, cosmetic, and food fields. This review summarizes the main research advances of O/W/O double emulsions over the past two decades. It mainly focuses on understanding the preparation strategies, stabilization mechanism, and potential applications of O/W/O double emulsions. Several emulsification strategies are discussed, including traditional two-step emulsification method, phase-inversion approach, membrane emulsification, and microfluidic emulsification. Further, the role of interfacial stabilizers and viscosity in the stability of O/W/O double emulsions will be discussed with a focus on synthetic emulsifiers, natural biopolymer sand solid particles for achieving this purpose. Additionally, analytical methods for evaluating the stability of O/W/O double emulsions, such as advanced microscopy, rheology, and labeling assay are reviewed taking into account potential limitations of these characterization techniques. Moreover, possible innovative food applications are highlighted, such as simulating fat substitutes to decrease the trans- or saturated fatty acid content and developing novel delivery and encapsulation systems. This review paves a solid way for the exploration of O/W/O double emulsions toward large-scale implementation within the food industry.

An in-depth multiphasic analysis of the chocolate production chain, from bean to bar, demonstrates the superiority of Saccharomyces cerevisiae over Hanseniaspora opuntiae as functional starter culture during cocoa fermentation.

Hanseniaspora opuntiae is a commonly found yeast species in naturally fermenting cocoa pulp-bean mass, which needed in-depth investigation. The present study aimed at examining effects of the cocoa isolate H. opuntiae IMDO 040108 as part of three different starter culture mixtures compared with spontaneous fermentation, regarding microbial community, substrate consumption, and metabolite production dynamics, including volatile organic compound (VOC) and phytochemical compositions, as well as compositions of the cocoa beans after fermentation, cocoa liquors, and chocolates. The inoculated H. opuntiae strain was unable to prevail over background yeasts present in the fermenting cocoa pulp-bean mass. It led to under-fermented cocoa beans after four days of fermentation, which was however reflected in higher levels of polyphenols. Cocoa fermentation processes inoculated with a Saccharomyces cerevisiae strain enhanced flavour production during the fermentation and drying steps, which was reflected in richer and more reproducible aroma profiles of the cocoa liquors and chocolates. Sensory analysis of the cocoa liquors and chocolates further demonstrated that S. cerevisiae led to more acidic notes compared to spontaneous fermentation, as a result of an advanced fermentation degree. Finally, different VOC profiles were found in the cocoa beans throughout the whole chocolate production chain, depending on the fermentation process.

Assessing the flavor of cocoa liquor and chocolate through instrumental and sensory analysis: a critical review.

The performance of appropriate instrumental and/or sensory analyses is essential to gain insights into the flavor profile of cocoa products. This three-part review is compiled of an overview of the most commonly used instrumental techniques to study cocoa liquor and chocolate flavor, their perception by a trained panel and the potential relationship between them. Each part is the result of a thorough literature study, principally focusing on the assumptions, features and limitations of these techniques. Reviewing of the literature revealed that cocoa matrix effects and methodology restraints were not always considered when instrumentally analyzing cocoa flavor. With respect to sensory analyses, various studies lacked reporting of accomplished trainings and performance of panelists. Moreover, a discrepancy was noticed in the descriptive flavor lexicon employed. Finally, when linking instrumental and sensory data, linear modeling is regularly applied, which might not always be appropriate. This review paper addresses the challenges associated with flavor assessment, intending to incite researchers to critically study cocoa flavor and apply standardized protocols and procedures.

Foliar water uptake does not contribute to embolism repair in beech (Fagus sylvatica L.).

BACKGROUND AND AIMS: Foliar water uptake has recently been suggested as a possible mechanism for the restoration of hydraulically dysfunctional xylem vessels. In this paper we used a combination of ecophysiological measurements, X-ray microcomputed tomography and cryo-scanning electron microscopy during a drought treatment to fully evaluate this hypothesis. KEY RESULTS: Based on an assessment of these methods in beech (Fagus sylvatica L.) seedlings we were able to (1) confirm an increase in the amount of hydraulically redistributed water absorbed by leaves when the soil water potential decreased, and (2) locate this redistributed water in hydraulically active vessels in the stem. However, (3) no embolism repair was observed irrespective of the organ under investigation (i.e. stem, petiole or leaf) or the intensity of drought. CONCLUSIONS: Our data provide evidence for a hydraulic pathway from the leaf surface to the stem xylem following a water potential gradient, but this pathway exists only in functional vessels and does not play a role in embolism repair for beech.

Development of a method to determine the SFC in the fat phase of emulsions using TD-NMR FID-CPMG deconvolution.

Fat crystallisation in emulsions is a complex process. One of the important parameters is the solid fat content (SFC). Up to now, there is no standardised method to measure the SFC in emulsions, let alone to determine the SFC of the fat inside droplets, thus avoiding the signal of the aqueous phase. This work evaluates the capabilities of deconvolution of the free induction decay (FID)-Carr-Purcell-Meiboom-Gill (CPMG) signal of emulsions. Three models were evaluated. The first model was a combination of a Gaussian function and a bi-exponential function (GBE model). The second model combined a Gaussian function with multiple exponential functions (GME model). The last model contained multiple Gaussian functions and multiple exponential functions (MGME model). The latter two models used a simplified CONTIN analysis. Based on the analysis of the determination coefficient R2 , the calculated water content and the estimated SFC of nonemulsified two-phase systems, the GBE model was selected to analyse the FID-CPMG signal of emulsified systems. However, the results obtained with the other models did not differ substantially, and hence, they could be used to obtain a full relaxation time distribution. When the GBE model was applied on different emulsion systems, no significant differences in estimated SFC of the fat phase were found, thus indicating that the emulsion formulation (i.e. water-in-oil [W/O], oil-in-water [O/W] or water-in-oil-in-water [W/O/W]) only had a minor effect on the SFC in the systems considered here.

Arabinoxylan, ß-glucan and pectin in barley and malt endosperm cell walls: a microstructure study using CLSM and cryo-SEM.

The architecture of endosperm cell walls in Hordeum vulgare (barley) differs remarkably from that of other grass species and is affected by germination or malting. Here, the cell wall microstructure is investigated using (bio)chemical analyses, cryogenic scanning electron microscopy (cryo-SEM) and confocal laser scanning microscopy (CLSM) as the main techniques. The relative proportions of ß-glucan, arabinoxylan and pectin in cell walls were 61, 34 and 5%, respectively. The average thickness of a single endosperm cell wall was 0.30 µm, as estimated by the cryo-SEM analysis of barley seeds, which was reduced to 0.16 µm after malting. After fluorescent staining, 3D confocal multiphoton microscopy (multiphoton CLSM) imaging revealed the complex cell wall architecture. The endosperm cell wall is composed of a structure in which arabinoxylan and pectin are colocalized on the outside, with ß-glucan depositions on the inside. During germination, arabinoxylan and ß-glucan are hydrolysed, but unlike ß-glucan, arabinoxylan remains present in defined cell walls in malt. Integrating the results, an enhanced model for the endosperm cell walls in barley is proposed.

Investigating the effect of different types of cocoa powder and stabilizers on suspension stability of cinnamon-cocoa drink.

Sedimentation of particles in cocoa drink is a technological challenge for the food industry. This study investigates the effect of different stabilizers (alginate, xanthan gum or carrageenan) on the suspension stability of cinnamon-cocoa drink made from 2 types of cocoa powder (natural or alkalized). Rheological and microstructural properties determination was used to examine the stabilization effect mechanism. The cocoa powder characteristic was investigated to study the correlation between cocoa powder properties and suspension stability. The results showed that xanthan gum is the most effective stabilizer to prevent particle sedimentation of the cinnamon-cocoa drink. Xanthan gum formed a network entrapping the particles. It increased the viscosity from 2.47 to 70.44 mPa s at a shear rate of 10/s. The drink formulated with alkalized cocoa powder has a better stability than that formulated with natural cocoa powder. However, at the concentration of 0.1% (w/v), xanthan gum could prevent sedimentation regardless the type of cocoa powder. The addition of xanthan gum up to 0.1% (w/v) had no significant effect on pH and antioxidant properties of the cinnamon-chocolate drink with a minor change in the lightness (L*) parameter. As such, the value of L*, pH, phenolic content and antioxidant activity of the cinnamon-cocoa drinks remained stable at around 22.5 ± 0.9, 7.2 ± 0.1, 0.31 ± 0.5 mg epicatechin equivalent /ml and 0.44 ± 0.3 mg tannic acid equivalent /ml, respectively. This study can be useful for the food industry to define a novel strategy to produce "ready-to-drink" cocoa-based beverage with prolonged suspension stability.

Antioxidant activity and quality attributes of white chocolate incorporated with Cinnamomum burmannii Blume essential oil.

White chocolate is often considered as an unhealthy product with low phenolic content and antioxidant activity since it does not contain cocoa liquor. In this study, investigation on the phytochemical composition of cinnamon essential oil as well as its potential use to improve the antioxidant activity of white chocolate were carried out. The effect of the essential oil incorporation on the quality attributes of white chocolate was also examined. The results show that cinnamon essential oil was rich in cinnamaldehyde and exhibited antioxidant activity. The incorporation of cinnamon essential oil at a level of 0.1% (w/w) increased the antioxidant activity of the white chocolate more than twofold without significant effect on its hardness, melting properties and colour. However, a slight alteration on the flow behaviour of the white chocolate was observed. This study clearly shows that natural cinnamon essential oil could be an alternative to synthetic additives in foods to improve their antioxidant activity.

Fat crystals: A tool to inhibit molecular transport in W/O/W double emulsions.

Water-in-oil-in-water (W/O/W) double emulsions are a promising technology for encapsulation applications of water soluble compounds with respect to functional food systems. Yet molecular transport through the oil phase is a well-known problem for liquid oil-based double emulsions. The influence of network crystallization in the oil phase of W/O/W globules was evaluated by NMR and laser light scattering experiments on both a liquid oil-based double emulsion and a solid fat-based double emulsion. Water transport was assessed by low-resolution NMR diffusometry and by an osmotically induced swelling or shrinking experiment, whereas manganese ion permeation was followed by means of T2 -relaxometry. The solid fat-based W/O/W globules contained a crystal network with about 80% solid fat. This W/O/W emulsion showed a reduced molecular water exchange and a slower manganese ion influx in the considered time frame, whereas its globule size remained stable under the applied osmotic gradients. The reduced permeability of the oil phase is assumed to be caused by the increased tortuosity of the diffusive path imposed by the crystal network. This solid network also provided mechanical strength to the W/O/W globules to counteract the applied osmotic forces.

Physical compatibility between wax esters and triglycerides in hybrid shortenings and margarines prepared in rice bran oil.

BACKGROUND: Wax esters contribute to the transformation of liquid oils into solid-like oleogel systems, which can act as alternatives for trans- and/or saturated fats in food products. The use of solely waxes reduces the solid content, consistency and sensory quality in the final products. Therefore, a combination of sunflower wax and palm fat in rice bran oil was created to accomplish the hybrid low-saturated shortenings and margarines with a compatible structure and lower amounts of saturated fats. RESULTS: During cooling of the hybrid shortenings, sunflower wax crystallized first and acted as nucleation sites for the crystallisation of palm fat. At 5 °C, a mixture of different crystal morphologies (α, ß', and ß crystals) existed in the hybrid shortening. In margarine processing, the hybrid samples were subjected to a simultaneous cooling-emulsification, in which sunflower wax crystallised first at the interface and adsorbed onto the water droplets. Based on the hardness measurements, the maximum amount of palm fat replaceable by 1.0%wt sunflower wax was up to 40% in shortenings and 25% in margarines. A higher amount of sunflower wax (2.5%wt) reduced up to 40% of saturated fats in the hybrid emulsions. CONCLUSION: The addition of 1.0%wt sunflower wax enhanced the solid content and network strength of hybrid palm-based shortenings. Sunflower wax helped to stabilise the water droplets inside the wax-based crystalline network without flocculation during shear-cooling. This research provides fundamental insight into the structuring of hybrid systems containing waxes, which could be interesting for the production of low-saturated fat products in the food industry. © 2017 Society of Chemical Industry.

The Contribution of Modern Margarine and Fat Spreads to Dietary Fat Intake.

The study of dietary fat consumption and its resultant effects on human health has been one of the most investigated topics in the field of human nutrition. Based on the results obtained from such studies, specific dietary recommendations on fat intake (both in terms of quantity and quality) have been established by health organizations around the globe. Among the various food industry sectors, the margarine manufacturers have also responded to these guidelines and now offer improved formulations with a desirable balance of fat contents and fat types. The main aim of this article is to provide an overview on how these modern margarines can contribute towards reaching the dietary guidelines relating to fat intake. In particular, the dietary recommendations with respect to the specific fatty acid types are comprehensively detailed along with an emphasis on the role of modern margarines in providing balanced fat types (more polyunsaturated fats, less saturated fats and a near-complete absence of trans fats) in the daily diet.

Biopolymer-based structuring of liquid oil into soft solids and oleogels using water-continuous emulsions as templates.

Physical trapping of a hydrophobic liquid oil in a matrix of water-soluble biopolymers was achieved using a facile two-step process by first formulating a surfactant-free oil-in-water emulsion stabilized by biopolymers (a protein and a polysaccharide) followed by complete removal of the water phase (by either high- or low-temperature drying of the emulsion) resulting in structured solid systems containing a high concentration of liquid oil (above 97 wt %). The microstructure of these systems was revealed by confocal and cryo-scanning electron microscopy, and the effect of biopolymer concentrations on the consistency of emulsions as well as the dried product was evaluated using a combination of small-amplitude oscillatory shear rheometry and large deformation fracture studies. The oleogel prepared by shearing the dried product showed a high gel strength as well as a certain degree of thixotropic recovery even at high temperatures. Moreover, the reversibility of the process was demonstrated by shearing the dried product in the presence of water to obtain reconstituted emulsions with rheological properties comparable to those of the fresh emulsion.

Polysaccharide-based oleogels prepared with an emulsion-templated approach.

The preparation and characterization of oleogels structured by using a combination of a surface-active and a non-surface-active polysaccharide through an emulsion-templated approach is reported. Specifically, the oleogels were prepared by first formulating a concentrated oil-in-water emulsion, stabilized with a combination of cellulose derivatives and xanthan gum, followed by the selective evaporation of the continuous water phase to drive the network formation, resulting in an oleogel with a unique microstructure and interesting rheological properties, including a high gel strength, G'>4000 Pa, shear sensitivity, good thixotropic recovery, and good thermostability.

Optimisation of enzymatic synthesis of cocoa butter equivalent from high oleic sunflower oil.

BACKGROUND: High oleic sunflower oil (HOSO) and a fatty acid (FA) mixture were inter-esterified in a solvent-free system catalysed by Lipozyme RM IM to produce a cocoa butter equivalent (CBE). The effects of reaction conditions on the percentage of saturate-oleoyl-saturate (SOS) and saturate-saturate-oleoyl (SSO) triacylglycerols (TAGs) were studied. The process was further optimised by response surface methodology. A five-factor response surface design was used to investigate the influences of the five major factors and their mutual relationships. The five factors were substrate ratio (A, FA/HOSO, mol mol⁻¹), enzyme load (B, wt% based on substrates), water content (C, wt% based on substrates), reaction temperature (D,°C) and reaction time (E, in hours) varying at three levels together with two star point levels. RESULTS: The highest yield (59.1% SOS) and lowest acyl migration (2.9% SSO) was obtained at 10% enzyme load, 1% water content, 1:7 substrate mole ratio, 65°C reaction temperature and 6 h reaction time. All the investigated factors except substrate ratio had significant effect on acyl migration. CONCLUSION: The quadratic response models sufficiently described the acidolysis reaction. All parameters had significant effect on the percentage of SOS TAGs. Based on the models, the reaction was optimised to obtain a maximum yield of SOS TAGs.

Terahertz time-domain transmission spectroscopy of water and hydrogel thin films: Extraction of optical parameters and application to agarose gel characterization.

Terahertz time-domain spectroscopy (THz-TDS) is an emerging optical technique that has potential applications in the characterization of (bio)materials. However, the complicated extraction of optical parameters from multi-layered and optically thin samples is a barrier towards its acceptance by applied scientists. Therefore, the aim of this work is to provide a straightforward approach for the extraction of the THz absorption coefficient and index of refraction profiles of aqueous thin films in a window-sample-window configuration, which is ubiquitous in many laboratories (i.e., sample in a cuvette). A numerical approach-based methodology that accounts for multiple layers, Fabry-Pérot effect, and sample thickness is elaborated which involves an optical interference model based on a tri-layer structure and a simple thickness estimation technique. This method was validated on water samples where a good agreement was found with the THz optical parameters of water reported in the literature, while the use of a commercial software resulted in erroneous optical parameters estimates when used without due regard to its limitations. A case study was then performed to demonstrate the ability of the proposed method to characterize agarose hydrogels with varying degree of sulfation. It was demonstrated that THz-TDS can provide insight into the hydration state of the agarose hydrogels, including the relative number of the hydrogen bonds between the hydroxyl moieties of water and the polysaccharide network which is perturbed by the presence of sulfate. The trend in the index of refraction profiles suggested microstructural differences between the agarose hydrogels, which were confirmed by visualizing the agarose network morphology using cryo-SEM imaging.

Edible insects as a novel source of lecithin: Extraction and lipid characterization of black soldier fly larvae and yellow mealworm.

Edible insects with high fat and phosphorus content are a potential novel source of lecithin, however, studies on their minor lipids are limited. In this study, lecithin was extracted from black soldier fly larvae and yellow mealworm. Herein, the effects of lecithin extraction method, matrix and ultrasound pretreatment were explored based on the fatty acid composition and phospholipid profile with soy lecithin as a reference. The use of a wet matrix and ultrasound pretreatment increased the extraction efficiency of total PLs from both insects. Insect lecithin contained a considerable amount of sphingomyelin compared to soy lecithin. In insect lecithin, a total of 47 glycerophospholipid and sphingomyelin molecular species, as well as four molecular species of fatty acyl esters of hydroxy fatty acid, were detected. This study is the first comprehensive investigation of insects as a new source of lecithin with applications in food, cosmetics and in the pharmaceutical industry.

Influence of calcium concentration on the re-assembly of sodium caseinate into casein micelles and on their renneting behavior.

Inducing the spontaneous aggregation from casein molecules (i.e. αs1, αs2, ß, and κ-casein) into re-assembled casein micelles (RCMs) through the addition of salts as an alternative to native casein micelles, has garnered increasing attention in recent years. In this investigation, re-assembled casein micelles were generated by adding varying amounts of calcium, phosphate, and citrate ions to a sodium caseinate dispersion. The formed micelles were further characterized in terms of particle size, optical density, and partitioning of calcium ions and caseins. Besides, their small-angle X-ray scattering (SAXS) profiles and renneting properties were evaluated. The observations revealed that the particle size and optical density of RCMs increased with the continuous addition of salts, while the micellar yield improved and could exceed 85 %. Moreover, the quantity of individual casein molecules that contributed to the creation of micelles was in concordance with their level of phosphorylation (i.e. αs2-casein > αs1-casein > ß-casein > κ-casein). Mineral analysis results and SAXS scattering profiles confirmed that the added calcium ions acted as cross-linkers and participated in the construction of calcium phosphate nanoclusters. The renneting ability of RCMs was primarily dependent upon the colloidal calcium content per gram of micellar casein.