Kinetic Model of Fluorescein Release through Bioprinted Polylactic Acid Membrane.

Polylactic acid (PLA)-based cylindrical membranes for the controlled release of fluorescein sodium salt (FS) were prepared by bioprinting on systems with an initial FS concentration of 0.003763 gdm-3 and 37.63 gdm-3, and the drug release process was monitored in a bath at 37 °C. Photographs, acquired at regular intervals during the process, revealed marked osmotic swelling of the polymer. Osmotic swelling consists in the enlargement of the polymer structure and due to the influx of water molecules across the membrane. The cylindrical PLA membrane starts to significantly swell once a certain threshold range is crossed. Important amounts of FS can dissolve under these radically changed circumstances, and the dissolved FS molecules are mobile enough to diffuse out of the cylinder, thus allowing drug release. As a matter of fact, in this investigation, we ascertained that polymer swelling promotes the mass transport phenomenon by altering the conditions for drug dissolution and diffusion, hence facilitating FS release after a specific lag time. Furthermore, in order to compare the release kinetics, the half-release time, t0.5, was taken into consideration. The data of this study evidence that, while increasing the initial concentration of FS by three orders of magnitude, the time parameter, t0.5, is only reduced by 5/6. In addition, the yield of the release process is drastically reduced due to the strong aggregation ability of the dye. Finally, it is demonstrated that a compressed exponential kinetic model fits the experimental data well despite the varying physical conditions.

Effects of Anionic Liposome Delivery of All-Trans-Retinoic Acid on Neuroblastoma Cell Differentiation.

All-trans-retinoic acid (ATRA) has long been known to affect cell growth and differentiation. To improve ATRA's therapeutic efficacy and pharmacodynamics, several delivery systems have been used. In this study, free ATRA and anionic-liposome-encapsulated ATRA were compared for their effects on SK-N-SH human neuroblastoma cell growth and differentiation. Anionic liposomes made of L-α-phosphatidylcholine (PC) and L-α-phosphatidic acid (PA), empty (PC-PA) and loaded with ATRA (PC-PA-ATRA), were characterized by dynamic light scattering (DLS) and electrophoretic mobility measurements, and drug entrapment efficiency (EE%) was measured to evaluate the applicability of the new colloidal formulation. The results of brightfield microscopy and cell growth curves indicated that ATRA, whether free or encapsulated, reduced growth and induced differentiation, resulting in SK-N-SH cells changing from epithelioid to neuronal-like morphologies, and producing a significant increase in neurite growth. To further characterize the neuro-differentiation of SK-N-SH cells, the expression of ßIII-Tubulin and synaptophysin and mitochondria localization were analyzed via immunofluorescence. Increased expression of neuronal markers and a peculiar localization of mitochondria in the neuritic extensions were apparent both in ATRA- and PC-PA-ATRA-differentiated cells. As a whole, our results strongly indicate that ATRA treatment, by any means, can induce the differentiation of parent SK-N-SH, and they highlight that its encapsulation in anionic liposomes increases its differentiation ability in terms of the percentage of neurite-bearing cells. Interestingly, our data also suggest an unexpected differentiation capability of anionic liposomes per se. This work highlights the importance of developing and carefully testing novel delivery nanocarriers, which are a necessary first "step" in the development of new therapeutic settings.

Alginate-Based Emulsions and Hydrogels for Extending the Shelf Life of Banana Fruit.

Edible coatings are used to extend the shelf life of various fruit, including bananas (Musa from the Musaceae family). After harvest, bananas reach the ripening and subsequent senescence phase. During senescence, the quality of the fruit deteriorates as it takes on a brown color and the tissue becomes soft. To extend the shelf life of such a fruit, effective methods to delay ripening are required. In this study, an alginate-based emulsion, i.e., an oil-in-water emulsion of lemongrass essential oil in alginate, was used to combine the mechanical properties of hydrocolloids with the water barrier properties of the oil phase. The emulsion was sprayed onto the whole fruit with an airbrush, and calcium chloride was added to promote gelling of the alginate. Compared to the uncoated fruit, coated bananas remained uniform in appearance (peel color) for longer, showed less weight loss, had a delay in the formation of total soluble solids, and in the consumption of organic acids. The shelf life of the coated fruit was extended by up to 11 days, at least 5 days more than uncoated bananas. Overall, the proposed coating could be suitable for reducing the global amount of food waste.

Antioxidative Properties of Fermented Soymilk Using Lactiplantibacillus plantarum LP95.

In recent times, there has been a growing consumer interest in replacing animal foods with alternative plant-based products. Starting from this assumption, for its functional properties, soymilk fermented with lactic acid bacteria is gaining an important position in the food industry. In the present study, soymilk was fermented with Lactiplantibacillus plantarum LP95 at 37 °C, without the use of stabilizers as well as thickeners and acidity regulators. We evaluated the antioxidant capacity of fermented soymilk along with its enrichment in aglycone isoflavones. The conversion of isoflavone glucosides to aglycones (genistein, glycitein, and daidzein) was analyzed together with antioxidant activity (ABTS) measurements, lipid peroxidation measurements obtained by a thiobarbituric acid reactive substance (TBARS) assay, and apparent viscosity measurements. From these investigations, soymilk fermentation using Lp. plantarum LP95 as a starter significantly increased isoflavones' transformation to their aglycone forms. The content of daidzein, glycitein, and genistein increased after 24 h of fermentation, reaching levels of 48.45 ± 1.30, 5.10 ± 0.16, and 56.35 ± 1.02 µmol/100 g of dry weight, respectively. Furthermore, the antioxidant activity increased after 6 h with a reduction in MDA (malondialdehyde). The apparent viscosity was found to increase after 24 h of fermentation, while it slightly decreased, starting from 21 days of storage. Based on this evidence, Lp. plantarum LP95 appears to be a promising candidate as a starter for fermented soymilk production.

Characterization, Sensory and Oxidative Stability Analysis of Vegetable Mayonnaise Formulated with Olive Leaf Vinegar as an Active Ingredient.

Development of novel food products represents a basic meeting point for health and business requirements. Mayonnaise sauce is well-suited to be a healthy and tasty dressing. In this study, mayonnaise was formulated by using unconventional ingredients, such as olive leaf vinegar (OLV), soybean/high oleic sunflower oil blend, and soymilk (as an egg substitute). An 18% alcoholic vinegar was used as the control sample. OLV is a rich source of bioactive substances, especially polyphenols and represents a possible way to enhance the olive oil by-product valorisation. For this new typology of vinegar an high level of phenolic compounds (7.2 mg/mL GAE), especially oleuropein (6.0 mg/mL oleuropein equivalent) was found. OLV mayonnaise had 57% fat, composed of 11%, 64%, and 23% saturated, monounsaturated, and polyunsaturated fatty acids, while linolenic acid was up to 1.7%. The phenol and oleuropein contents were 68 and 52 mg/100 g, respectively. Sensory panellists expressed a moderate overall acceptability for both samples but attested more distinctive and positive sensations for the colour, odour, and taste attributes of OLV mayonnaise. Finally, oxidative stability and shelf life were better in OLV mayonnaise than in the control. Specifically, the peroxide value remained low (around 4.5 meqO2/kg) after 12 months of storage at room and low (4 °C) temperatures.

Effect of α-Amylase on the Structure of Chia Seed Mucilage.

Thanks to its nutritional and mechanical properties, chia seed mucilage is becoming increasingly popular in the food industry as a small biomolecule. The mechanical properties of an ingredient are a key element for food appreciation during chewing. Therefore, with this study, we explore for the first time the structural changes that chia seed mucilage undergoes when treated with α-amylase, the most abundant enzyme in human saliva. First, rheological time-sweep tests were performed on samples with different enzyme and constant chia mucilage concentrations. Then, the effect of increasing the chia mucilage concentration at a constant enzyme content was investigated. The results show that structural changes occur after enzyme treatment. Rheological measurements show a thickening of the material with an increase in the elastic modulus depending on the concentrations of α-amylase and chia used. This effect is attributed to the release and aggregation of insoluble fibrous aggregates that naturally form the mucilage after the cleavage of the α-1,4-glucoside bond between the α-D-glucopyranose residue and the second ß-D-xylopyranose residue by α-amylase. Thus, our data suggest an α-amylase-mediated restructuring of the chia mucilage network that could have implications for the commercial processing of this material.

Progress in Colloid Delivery Systems for Protection and Delivery of Phenolic Bioactive Compounds: Two Study Cases-Hydroxytyrosol and Curcumin.

Insufficient intake of beneficial food components into the human body is a major issue for many people. Among the strategies proposed to overcome this complication, colloid systems have been proven to offer successful solutions in many cases. The scientific community agrees that the production of colloid delivery systems is a good way to adequately protect and deliver nutritional components. In this review, we present the recent advances on bioactive phenolic compounds delivery mediated by colloid systems. As we are aware that this field is constantly evolving, we have focused our attention on the progress made in recent years in this specific field. To achieve this goal, structural and dynamic aspects of different colloid delivery systems, and the various interactions with two bioactive constituents, are presented and discussed. The choice of the appropriate delivery system for a given molecule depends on whether the drug is incorporated in an aqueous or hydrophobic environment. With this in mind, the aim of this evaluation was focused on two case studies, one representative of hydrophobic phenolic compounds and the other of hydrophilic ones. In particular, hydroxytyrosol was selected as a bioactive phenol with a hydrophilic character, while curcumin was selected as typical representative hydrophobic molecules.

Alginate-Assisted Lemongrass (Cymbopogon nardus) Essential Oil Dispersions for Antifungal Activity.

The use of natural compounds as food preservatives is becoming increasingly popular as it is perceived positively by consumers. Among these substances, essential oils have attracted great interest owing to their antioxidant and antimicrobial properties. However, several challenges impair the use of essential oils in food products, such as their degradation or loss during food processing and storage, the strong aroma, even at low concentrations, which may negatively affect the sensory characteristics of food. In this context, the development of nanoformulations able to stabilize essential oils may represent a smart solution to this issue. The aim of the study was to evaluate the efficiency of alginate-based nanoformulations enriched with lemongrass (Cymbopogon nardus) essential oil (LEO) and Tween 80 against several fungi namely Penicillium expansus, Aspergillus niger and Rhizopus spp. Firstly, the flow behavior of systems at different concentrations of alginate (1%, 2% and 3% w/w) were studied. Then, emulsion-based nanoformulations at different concentrations of lemongrass essential oil in the range of 0-2% w/w were stabilized by a fixed amount of Tween 80, characterized and tested for their antifungal activity. Our results showed that the best nanoformulation able to inhibit Rhizopus spp., Penicillium expansum and Aspergillus niger, for at least 10 days, was constituted by 1% alginate/1.5% LEO/1% Tween 80. Hence, the incorporation of essential oil into nanoformulation systems may represent a valid alternative to overcome the disadvantages that limit the commercial application of essential oils.

Rheological and Nutritional Assessment of Dysphagia-Oriented New Food Preparations.

Dysphagia that involves difficulty swallowing food and liquids is a symptom of different diseases. In some cases, patients who experience this symptom should be fed with modified consistency foods. Dysphagia is often accompanied by malnutrition and dehydration and an interesting approach to dealing with these conditions is to provide patients with nutrient-rich foods. In this study, two new food formulations for dysphagia patients are proposed: a cereal-based protein meal and a vegetable cream. The nutritional and rheological characteristics of the two innovative preparations were assessed and compared with those of commercial products. The proposed protein meal formulations meet the criteria for the "high protein" claim and the vegetable cream meets those of the "source of fiber" claim. The rheological investigation revealed that the flow properties of the innovative formulations were comparable to those of the commercial ones. Based on these preliminary outcomes, this investigation represents an interesting perspective potentially valuable to enlarge the offer of possibilities for people suffering from swallowing disorders.

Insights into Mechanical Behavior and Biological Properties of Chia Seed Mucilage Hydrogels.

In this contribution we report insights on the rheological properties of chia (Salvia hispanica) seed mucilage hydrogels. Creep experiments performed in steady state conditions allowed calculation of Newtonian viscosities for chia hydrogels with different polymer concentration, pointing at inter-chain interactions as the main responsible for the different behavior toward network slipping under constant stress. A combination of oscillatory frequency and stress sweep tests highlighted a moderate effect of temperature in influencing hydrogel mechanics. The latter results prompted us to investigate potential biological functions for this set of biomaterials. Lactate Dehydrogenase assay proved the lack of cytotoxicity of chia suspensions toward Human Mesenchymal Stem Cells from adipose tissue used here as a cell model. Differentiation experiments were finally undertaken to verify the influence of chia samples on osteo-induction triggered by chemical differentiation factors. Alkaline Phosphatase enzyme activity assay and Alizarin red staining demonstrated that chia mucilage did not alter in vitro stem cell differentiation. Collectively, this set of experiments revealed an almost inert role associated with chia suspensions, indicating a possible application of chia-based networks as scaffold models to study osteogenesis in vitro.

Oral delivery of all-trans retinoic acid mediated by liposome carriers.

All-trans retinoic acid (ATRA) is a molecule that finds wide applications in medicine. Connection between cancer cell proliferation and ATRA is a well-established item. Driven by the potential applications of liposomes in stabilizing and protecting therapeutic compounds thus enabling effective delivery of encapsulated compounds, recent research efforts have been directed to understanding mechanisms of oral delivery through the gastrointestinal tract. The surface charge of the liposome bilayers can modify the interactions between the aggregates and the gastrointestinal fluids. Here, we investigated the ability of cationic and anionic liposomes to encapsulate, protect and deliver ATRA in an in-vitro digestion process as a different oral administration route. Stability and encapsulation efficiency of ATRA in negatively and positively charged liposomes enriched with α-tocopherol were investigated by means of UV-vis spectroscopy, dynamic light scattering and ζ-potential. The applicability of the carriers was tested by means of an in-vitro digestion procedure allowing for the measurement of the bioavailability of ATRA. From this study evidence was provided that the water insoluble molecules, ATRA and α-tocopherol are intercalated in liposome membranes regardless of the surface charge of the vesicle bilayers. Comparisons between cationic and anionic liposomes incorporating retinoic acid show differences in bioavailability. The cationic vesicles are preferable for a larger amount of ATRA bioavailability, which can be understood from electrostatic interactions. Thus ATRA is ionized in a wide range of pHs but protonated in anionic vesicles.

On the Mechanism of Genipin Binding to Primary Amines in Lactose-Modified Chitosan at Neutral pH.

The present manuscript deals with the elucidation of the mechanism of genipin binding by primary amines at neutral pH. UV-VIS and CD measurements both in the presence of oxygen and in oxygen-depleted conditions, combined with computational analyses, led to propose a novel mechanism for the formation of genipin derivatives. The indications collected with chiral and achiral primary amines allowed interpreting the genipin binding to a lactose-modified chitosan (CTL or Chitlac), which is soluble at all pH values. Two types of reaction and their kinetics were found in the presence of oxygen: (i) an interchain reticulation, which involves two genipin molecules and two polysaccharide chains, and (ii) a binding of one genipin molecule to the polymer chain without chain-chain reticulation. The latter evolves in additional interchain cross-links, leading to the formation of the well-known blue iridoid-derivatives.

Surfactant Interactions with Protein-Coated Surfaces: Comparison between Colloidal and Macroscopically Flat Surfaces.

Surface interactions with polymers or proteins are extensively studied in a range of industrial and biomedical applications to control surface modification, cleaning, or biofilm formation. In this study we compare surfactant interactions with protein-coated silica surfaces differing in the degree of curvature (macroscopically flat and colloidal nanometric spheres). The interaction with a flat surface was probed by means of surface plasmon resonance (SPR) while dynamic light scattering (DLS) was used to study the interaction with colloidal SiO2 (radius 15 nm). First, the adsorption of bovine serum albumin (BSA) with both SiO2 surfaces to create a monolayer of coating protein was studied. Subsequently, the interaction of these BSA-coated surfaces with a non-ionic surfactant (a decanol ethoxylated with an average number of eight ethoxy groups) was investigated. A fair comparison between the results obtained by these two techniques on different geometries required the correction of SPR data for bound water and DLS results for particle curvature. Thus, the treated data have excellent quantitative agreement independently of the geometry of the surface suggesting the formation of multilayers of C10PEG over the protein coating. The results also show a marked different affinity of the surfactant towards BSA when the protein is deposited on a flat surface or individually dissolved in solution.

Binary Solutions of Hyaluronan and Lactose-Modified Chitosan: The Influence of Experimental Variables in Assembling Complex Coacervates.

A miscibility study between oppositely charged polyelectrolytes, namely hyaluronic acid and a lactose-modified chitosan, is here reported. Experimental variables such as polymers' weight ratios, pH values, ionic strengths and hyaluronic acid molecular weights were considered. Transmittance analyses demonstrated the mutual solubility of the two biopolymers at a neutral pH. The onset of the liquid-liquid phase separation due to electrostatic interactions between the two polymers was detected at pH 4.5, and it was found to be affected by the overall ionic strength, the modality of mixing and the polymers' weight ratio. Thorough Dynamic Light Scattering (DLS) measurements were performed to check the quality of the formed coacervates by investigating their dimensions, homogeneity and surface charge. The whole DLS results highlighted the influence of the hyaluronic acid molecular weight in affecting coacervates' dispersity and size.

Enhanced Curcumin Bioavailability through Nonionic Surfactant/Caseinate Mixed Nanoemulsions.

The potential of heterogeneous systems like oil-in-water (O/W) nanoemulsions is exploited as an oral delivery system for curcumin, a natural lipophilic compound with numerous health benefits. Two types of O/W nanoemulsions, one stabilized by sodium caseinate (Cas-O/W), a surface-active and emulsifying protein, and the other stabilized by a blend of caseinate and Tween 20 (Mix-O/W), were loaded with the bioactive compound and tested through a simulated gastrointestinal digestion process to evaluate their effects on delivering of curcumin. It was first demonstrated that the amount of curcumin solubilized through Mix-O/W nanoemulsion was higher than that in Cas-O/W nanoemulsion. Cas-O/W nanoemulsions, indeed, at their best, solubilized about 55 µg/mL of curcumin while Mix-O/W nanoemulsions reached a curcumin concentration around 180 µg/mL. Furthermore, for both the systems an increase of curcumin loading capacity was recorded with the rise of incubation temperature. Finally, after the in vitro simulated digestion process, the potential curcumin bioavailability was evaluated and the data suggested that Mix-O/W nanoemulsions provided more than twice the amount of curcumin compared to Cas-O/W nanoemulsions. On balance, the outcomes of this investigation demonstrated that the mixed emulsifier system offered a higher amount of lipophilic compound with a low fat intake compared to nanoemulsions stabilized by sodium caseinate. PRACTICAL APPLICATION: The outcomes of this study allow the recognition of the protein/surfactant-stabilized nanoemulsions as a suitable solution to deliver curcumin. The nanoemulsions proposed here provide a high intake of curcumin, a lipophilic compound, with low fat content. The use of such delivery systems helps to overcome limits in oral bioavailability related with the scarce solubility of some compounds in food preparations and beverages.

Quality Control of Fresh-Cut Apples after Coating Application.

The growing demand for ready-to-eat fresh fruits has led to set-up appropriate strategies for preserving fruit quality and freshness of such commodities. To slow down the deterioration events such as respiration, moisture loss and enzymatic activity, ready-to-eat products should be protected with an edible film. A suitable coating should combine hydrophilic and hydrophobic features to ensure good mechanical and gas barrier properties. Alginate/essential oil nanoformulations, one with low and the other with high oil content, here proposed to protect apple pieces during storage, were first characterized through dynamic light scattering and rheology. The effect of the application of the nanoformulations on the quality parameters of apples stored at 4 °C was considered by evaluating weight loss, pH and titratable acidity, total phenols content and the fruit appearance during storage. Mainly on the basis of pH and titratable acididty variation, the nanoformulation with low oil content resulted eligible for preserving the quality of fresh-cut apple pieces during storage.

Rheological Characterization of Hydrogels from Alginate-Based Nanodispersion.

The interest toward alginate and nanoemulsion-based hydrogels is driven by the wide potential of application. These systems have been noticed in several areas, ranging from pharmaceutical, medical, coating, and food industries. In this investigation, hydrogels prepared through in situ calcium ion release, starting from lemongrass essential oil nanodispersions stabilized in alginate aqueous suspensions in the presence of the nonionic surfactant Tween 80, were evaluated. The hydrogels prepared at different concentrations of oil, alginate, and calcium were characterized through rheological tests. Flow curves demonstrate that the hydrogels share shear thinning behavior. Oscillatory tests showed that the strength of the hydrogel network increases with the crosslinker increase, and decreases at low polymer concentrations. The hydrogels were thixotropic materials with a slow time of structural restoration after breakage. Finally, by analyzing the creep recovery data, the hydrogel responses were all fitted to the Burger model. Overall, it was demonstrated that the presence of essential oil in the proposed hydrogels does not affect the mechanical characteristics of the materials, which are mainly influenced by the concentration of polymer and calcium as a crosslinker.

Determination of bisphenol A in red wine using a double vortex-ultrasound-assisted microextraction assay: Role of the interfacial properties.

Bisphenol A (BPA) is a synthetic compound broadly used in medical devices as well as in packaging of food and drinks. Recently, BPA toxicity has become of concern to environmental public health. Red wine that is susceptible to BPA contamination is an alcoholic beverage made from yeast fermentation of grapes in the presence of grape skins so as to extract phenolic compounds. The aim of this study was to validate an efficient, low cost, and time-saving method for BPA determination in red-wine beverage. To this end, a rapid and simple microextraction method is here proposed consisting in liquid-liquid separation assisted by a vortex-ultrasound-vortex procedure combined with gas chromatographic analysis (GC-Fid or GC-IT/MS). By means of a comparative study between real red-wine matrix and synthetic hydroalcoholic solutions, different parameters related to the microextraction steps were investigated. The minimal amount of extraction solvent for a given volume of sample was calculated for both the systems. It was demonstrated that for red-wine matrix, the extent of phase separation is strongly affected by some wine constituents and that separation can be tuned by varying the amount of the extraction solvent. This double vortex-ultrasound-assisted method achieved high recovery of BPA and enrichment factor compared with other microextraction methods.

Burst-Like Transcription of Mutant and Wildtype MYH7-Alleles as Possible Origin of Cell-to-Cell Contractile Imbalance in Hypertrophic Cardiomyopathy.

Hypertrophic Cardiomyopathy (HCM) has been related to many different mutations in more than 20 different, mostly sarcomeric proteins. While development of the HCM-phenotype is thought to be triggered by the different mutations, a common mechanism remains elusive. Studying missense-mutations in the ventricular beta-myosin heavy chain (ß-MyHC, MYH7) we hypothesized that significant contractile heterogeneity exists among individual cardiomyocytes of HCM-patients that results from cell-to-cell variation in relative expression of mutated vs. wildtype ß-MyHC. To test this hypothesis, we measured force-calcium-relationships of cardiomyocytes isolated from myocardium of heterozygous HCM-patients with either ß-MyHC-mutation Arg723Gly or Arg200Val, and from healthy controls. From the myocardial samples of the HCM-patients we also obtained cryo-sections, and laser-microdissected single cardiomyocytes for quantification of mutated vs. wildtype MYH7-mRNA using a single cell RT-qPCR and restriction digest approach. We characterized gene transcription by visualizing active transcription sites by fluorescence in situ hybridization of intronic and exonic sequences of MYH7-pre-mRNA. For both mutations, cardiomyocytes showed large cell-to-cell variation in Ca++-sensitivity. Interestingly, some cardiomyocytes were essentially indistinguishable from controls what might indicate that they had no mutant ß-MyHC while others had highly reduced Ca++-sensitivity suggesting substantial fractions of mutant ß-MyHC. Single-cell MYH7-mRNA-quantification in cardiomyocytes of the same patients revealed high cell-to-cell variability of mutated vs. wildtype mRNA, ranging from essentially pure mutant to essentially pure wildtype MYH7-mRNA. We found 27% of nuclei without active transcription sites which is inconsistent with continuous gene transcription but suggests burst-like transcription of MYH7. Model simulations indicated that burst-like, stochastic on/off-switching of MYH7 transcription, which is independent for mutant and wildtype alleles, could generate the observed cell-to-cell variation in the fraction of mutant vs. wildtype MYH7-mRNA, a similar variation in ß-MyHC-protein, and highly heterogeneous Ca++-sensitivity of individual cardiomyocytes. In the long run, such contractile imbalance in the myocardium may well induce progressive structural distortions like cellular and myofibrillar disarray and interstitial fibrosis, as they are typically observed in HCM.

Effect of the coexistence of sodium caseinate and Tween 20 as stabilizers of food emulsions at acidic pH.

In the present investigation the properties of edible nanoemulsions were studied. Sodium caseinate represents a good candidate for food emulsion preparations thanks to its surface-active properties and because it is perceived as a natural product by consumers. Nevertheless, it is very sensitive to acidic pH close to its isoelectric point and, if used as emulsion stabilizer, this aspect can negatively affect the emulsion stability. In order to prevent this drawback, sodium caseinate was used in combination with a non-ionic surfactant (Tween 20) as emulsifier of oil/water nanoemulsions. For these reasons, nanoemulsions stabilized by Tween 20, sodium caseinate and by a blend of the two emulsifiers were studied and compared according to their response to pH variations. Nanoemulsions were characterized for size of the dispersed phase with variation of time and temperature, for their rheological properties, for surface charge as a function of pH and for protein fluorescence. Noticeably, it was ascertained that, at pH close to caseinate isoelectric point, emulsions stabilized with the blend of caseinate and Tween 20 were more stable, compared with emulsions stabilized only with sodium caseinate. Such behavior was explained according to the composition of the emulsifiers at the oil/water interface where, at acidic pH, the presence of Tween 20 ensured the steric stabilization thus improving the role of sodium caseinate as emulsion stabilizer.