Phage Lytic Protein CHAPSH3b Encapsulated in Niosomes and Gelatine Films.

Antimicrobial resistance (AMR) has emerged as a global health challenge, sparking worldwide interest in exploring the antimicrobial potential of natural compounds as an alternative to conventional antibiotics. In recent years, one area of focus has been the utilization of bacteriophages and their derivative proteins. Specifically, phage lytic proteins, or endolysins, are specialized enzymes that induce bacterial cell lysis and can be efficiently produced and purified following overexpression in bacteria. Nonetheless, a significant limitation of these proteins is their vulnerability to certain environmental conditions, which may impair their effectiveness. Encapsulating endolysins in vesicles could mitigate this issue by providing added protection to the proteins, enabling controlled release, and enhancing their stability, particularly at temperatures around 4 °C. In this work, the chimeric lytic protein CHAPSH3b was encapsulated within non-ionic surfactant-based vesicles (niosomes) created using the thin film hydrating method (TFH). These protein-loaded niosomes were then characterized, revealing sizes in the range of 30-80 nm, zeta potentials between 30 and 50 mV, and an encapsulation efficiency (EE) of 50-60%. Additionally, with the objective of exploring their potential application in the food industry, these endolysin-loaded niosomes were incorporated into gelatine films. This was carried out to evaluate their stability and antimicrobial efficacy against Staphylococcus aureus.

Rapid and sensitive detection of E. coli O157:H7 by lateral flow immunoassay and silver enhancement.

The aim of this study was to develop a sensitive lateral flow immunoassay (LFIA) for the rapid detection of Escherichia coli (E. coli) O157:H7, a pathogen contributor to diseases and fatalities worldwide. Au nanoparticles with high stability, uniform size, and shape were synthesized and coated with heterobifunctional PEG polymer with carboxyl groups, and they were bioconjugated to be used as label in sandwich-LFIA. Then, a silver enhancement strategy was developed as an accessible, rapid, and cost-effective approach for signal amplification to reduce the limit of detection (LOD). The optimal results were achieved when a solution of silver nitrate and hydroquinone/citrate buffer was added to the strips for 4 min. This led to a decrease in the visual LOD from 2 × 106 (CFU mL-1) to 2 × 103 (CFU mL-1), resulting in a threefold improvement in sensitivity compared to the conventional LFIA system. The specificity of the system was evaluated by using non-target bacteria (E. coli BL21 and E. coli T515) and its reliability was determined by testing commercial food samples (milk, tap water, and orange juice), demonstrating its effectiveness for quickly detecting pathogenic bacteria in food products.

Fe3O4@Au Core-Shell Magnetic Nanoparticles for the Rapid Analysis of E. coli O157:H7 in an Electrochemical Immunoassay.

Escherichia coli (E. coli) O157:H7 is a pathogenic bacterium that causes serious toxic effects in the human gastrointestinal tract. In this paper, a method for its effective analytical control in a milk sample was developed. To perform rapid (1 h) and accurate analysis, monodisperse Fe3O4@Au magnetic nanoparticles were synthesized and used in an electrochemical sandwich-type magnetic immunoassay. Screen-printed carbon electrodes (SPCE) were used as transducers, and electrochemical detection was performed by chronoamperometry using a secondary horseradish peroxidase-labeled antibody and 3,3',5,5'-tetramethylbenzidine. This magnetic assay was used to determine the E. coli O157:H7 strain in the linear range from 20 to 2 × 106 CFU/mL, with a limit of detection of 20 CFU/mL. The selectivity of the assay was tested using Listeria monocytogenes p60 protein, and the applicability of the assay was assessed by analyzing a commercial milk sample, demonstrating the usefulness of the synthesized nanoparticles in the developed magnetic immunoassay.

Synthesis of controlled-size starch nanoparticles and superparamagnetic starch nanocomposites by microemulsion method.

In this study, a synthesis process based on the microemulsion method (ME) was developed with the aim to produce controlled-size starch nanoparticles (SNPs). Several formulations were tested for the preparation of the W/O microemulsions varying the organic/aqueous phase ratios and co-stabilizers concentrations. SNPs were characterized in terms of size, morphology, monodispersity and crystallinity. Spherical shape particles with mean sizes 30-40 nm were prepared. The method was then used to simultaneously synthesize SNPs and iron oxide nanoparticles with superparamagnetic properties. Starch-based nanocomposites with superparamagnetic properties and controlled size were obtained. Therefore, the microemulsion method developed could be considered an innovative technology for the design and development of novel functional nanomaterials. The starch-based nanocomposites were evaluated in terms of morphology and magnetic properties, and they are being considered as promising sustainable nanomaterials for different biomedical applications.

Nanovesicles as Vanillin Carriers for Antimicrobial Applications.

Vanillin is a natural compound easily extracted from plants. It has neuroprotective, anti-carcinogenic, antioxidant, antimicrobial, and anti-biofilm properties. It also presents high volatility, high hydrophilicity, and low bioavailability. Nanomaterials can be used to improve pharmacodynamics, solubility, and stability and to enhance pharmacokinetics. In this work, non-ionic surfactant vesicles were synthesized as vanillin carriers: neutral niosomes formed by Span60 and cholesterol, positive charged niosomes formulated with cetyltrimethylammonium bromide (CTAB), and negatively charged niosomes formulated with sodium dodecyl sulfate (SDS). Niosomes synthesis was carried out with two commonly used methods: thin film hydration (TFH) and ethanol injection method (EIM). The niosomes synthesized were used to prepare two different materials: (i) a powder containing the lyophilized noisome with vanillin systems and (ii) a gelatin matrix film containing niosomes with vanillin. Lyophilization was carried out using maltodextrin as a cryoprotectant. The lyophilization of colloidal structures allows for storage at room temperature for long periods of time, keeping their organoleptic characteristics invariable. Niosomes were characterized before and after the lyophilization process in terms of morphological characterization, size, polydispersity index (PDI), and zeta potential. Moreover, niosomes cargo was evaluated by calculating the encapsulation efficiency (EE) and loading capacity (LC). Results showed that the use of the TFH method allowed us to obtain niosomes of 255 nm with high EE (up to 40%) and LC values higher than EIM. The lyophilization process decreased the LC of the vesicles prepared, but this decrease was mitigated by up to 20% when ionic surfactants were used on the membrane bilayer. Gelatin films are biodegradable materials suitable for food packing applications. The incorporation of a natural compound with antimicrobial activity would be a clear advantage for such an application. The films prepared were characterized in terms of morphology, water solubility, color, and transparency. Niosomes synthesized by thin film hydration had better chemical and physical properties to load vanillin. Especially in the case of application in films, niosomes with a negative charge, formed by SDS, and vanillin loaded gave better mechanical and chemical characteristics to the film.

Stability of Non-Ionic Surfactant Vesicles Loaded with Rifamycin S.

These days, the eradication of bacterial infections is more difficult due to the mechanism of resistance that bacteria have developed towards traditional antibiotics. One of the medical strategies used against bacteria is the therapy with drug delivery systems. Non-ionic vesicles are nanomaterials with good characteristics for encapsulating drugs, due to their bioavailability and biodegradability, which allow the drugs to reach the specific target and reduce their side effects. In this work, the antibiotic Rifamycin S was encapsulated. The rifamycin antibiotics family has been widely used against Mycobacterium tuberculosis, but recent studies have also shown that rifamycin S and rifampicin derivatives have bactericidal activity against Staphylococcus epidermidis and Staphylococcus aureus. In this work, a strain of S. aureus was selected to study the antimicrobial activity through Minimum Inhibitory Concentration (MIC) assay. Three formulations of niosomes were prepared using the thin film hydration method by varying the composition of the aqueous phase, which included MilliQ water, glycerol solution, or PEG400 solution. Niosomes with a rifamycin S concentration of 0.13 µg/g were satisfactorily prepared. Nanovesicles with larger size and higher encapsulation efficiency (EE) were obtained when using glycerol and PEG400 in the aqueous media. Our results showed that niosomes consisting of an aqueous glycerol solution have higher stability and EE across a diversity of temperatures and pHs, and a lower MIC of rifamycin S against S. aureus.

A Dynamic, In Vitro BBB Model to Study the Effects of Varying Levels of Shear Stress.

The blood-brain barrier (BBB) consists of a tight network of blood capillaries in the brain that separate the circulatory system from the central nervous system. Its particular properties are based on the dynamic interaction between cerebral endothelial cells and other surrounding cells, especially astrocytes. We have designed and synthesized a three-dimensional scaffold that recapitulates the main hallmarks of the BBB extracellular matrix and serves as a platform to co-culture human brain microvascular endothelial cells and human cortical astrocytes. The scaffold can be exposed to flow, thereby allowing the study of flow-mediated pathways at the BBB.

The Effect of Precipitation pH on Protein Recovery Yield and Emulsifying Properties in the Extraction of Protein from Cold-Pressed Rapeseed Press Cake.

Rapeseed is the second most cultivated oilseed after soybean and is mainly used to produce vegetable oil. The by-product rapeseed press cake is rich in high-quality proteins, thus having the possibility of becoming a new plant protein food source. This study aimed to investigate how the precipitation pH affects the protein yield, protein content, and emulsifying properties when industrially cold-pressed rapeseed press cake is used as the starting material. Proteins were extracted under alkaline conditions (pH 10.5) with an extraction coefficient of 52 ± 2% followed by precipitation at various pH (3.0-6.5). The most preferred condition in terms of process efficiency was pH 4.0, which is reflected in the zeta potential results, where the proteins' net charge was 0 at pH 4.2. pH 4.0 also exhibited the highest protein recovery yield (33 ± 0%) and the highest protein concentration (64 ± 1%, dry basis). Proteins precipitated at pH 6.0-6.5 stabilized emulsions with the smallest initial droplet size, although emulsions stabilized by rapeseed protein precipitated at pH 5.0-6.0 showed the highest emulsion stability at 37 °C for 21 days, with a limited layer of free oil. Overall, emulsion stabilized by protein precipitated at pH 5.0 was the most stable formulation, with no layer of free oil after 21 days of incubation.

Encapsulation of Pomegranate Peel Extract (Punica granatum L.) by Double Emulsions: Effect of the Encapsulation Method and Oil Phase.

Pomegranate peel is an agro-industrial waste that can be used as source of punicalagin, a polyphenolic compound with several beneficial effects on health. Since, once extracted, punicalagin is prone to degradation, its encapsulation by double emulsions can be an alternative to protect the active compound and control its release. The aim of this investigation was to evaluate the feasibility of encapsulating pomegranate peel extract (PPE) in double emulsions using different types of oils (castor, soybean, sunflower, Miglyol and orange) in a ratio of 70:30 (oil:PPE) and emulsification methods (direct membrane emulsification and mechanical agitation), using polyglycerol polyricinoleate (PGPR) and Tween 80 as lipophilic and hydrophilic emulsifiers, respectively. Direct membrane emulsification (DME) led to more stable emulsions during storage. Droplet size, span values, morphology and encapsulation efficiency (EE) were better for double emulsions (DEs) prepared by DME than for mechanical agitation (MA). DEs formulated using Miglyol or sunflower oil as the oily phase could be considered as suitable food grade systems to encapsulate punicalagin with concentrations up to 11,000 mg/L of PPE.

From process effluents to intestinal health promotion: Developing biopolymer-whey liposomes loaded with gingerol to heal intestinal wounds and neutralize oxidative stress.

As a sustainable strategy to valorize the main effluent of the cheese industry and potent environmental pollutant, whey, several biopolymer-whey vesicles loaded with gingerol were tailored for counteracting intestinal oxidative stress and boosting wound healing. An eco-friendly method was used to combine whey with four different water-dispersible biopolymers (xanthan gum, tragacanth, Arabic gum and sodium alginate), phospholipid and a natural antioxidant (gingerol). The results of cryogenic transmission microscopy and dynamic light scattering indicated that the vesicles were mostly unilamellar and small in size (∼100 nm) with low polydispersity index, high negative zeta potential and ability to entrap a high amount of gingerol (up to 94%). The vesicles could maintain their structures in acidic and neutral media and Turbiscan® technology confirmed their stability during the storage. Vesicles prepared with whey and tragacanth exhibited the highest capability to protect intestinal cells from damages induced by hydrogen peroxide. When Arabic and tragacanth gums were added to the whey vesicles, the closure rate of the scratched area was fast and no trace of the wound was observed after 72 h of treatment. These promising findings could open a new horizon in the application of whey in nanomedicine for the treatment of intestinal damages.

Lipid-Polymer Hybrids Encapsulating Iron-Oxide Nanoparticles as a Label for Lateral Flow Immunoassays.

The feasibility of using Superparamagnetic Iron Oxide Nanoparticles (SPIONs) encapsulated by lipid-polymer nanoparticles as labels in lateral flow immunoassays (LFIA) was studied. First, nanoparticles were synthesized with average diameters between 4 and 7 (nm) through precipitation in W/O microemulsion and further encapsulated using lipid-polymer nanoparticles. Systems formulated were characterized in terms of size and shape by DLS (Nanozetasizer from Malvern) and TEM. After encapsulation, the average size was around (≈20 and 50 nm). These controlled size agglomerates were tested as labels with a model system based on the biotin-neutravidin interaction. For this purpose, the encapsulated nanoparticles were conjugated to neutravidin using the carbodiimide chemistry, and the LFIA was carried out with a biotin test line. The encapsulated SPIONs showed that they could be promising candidates as labels in LFIA test. They would be useful for immunomagnetic separations, that could improve the limits of detection by means of preconcentration.

The Effect of pH and Storage Temperature on the Stability of Emulsions Stabilized by Rapeseed Proteins.

Rapeseed press cake (RPC), the by-product of rapeseed oil production, contains proteins with emulsifying properties, which can be used in food applications. Proteins from industrially produced RPC were extracted at pH 10.5 and precipitated at pH 3 (RPP3) and 6.5 (RPP6.5). Emulsions were formulated at three different pHs (pH 3, 4.5, and 6) with soy lecithin as control, and were stored for six months at either 4 °C or 30 °C. Zeta potential and droplet size distribution were analyzed prior to incubation, and emulsion stability was assessed over time by a Turbiscan instrument. Soy lecithin had significantly larger zeta potential (-49 mV to 66 mV) than rapeseed protein (-19 mV to 20 mV). Rapeseed protein stabilized emulsions with smaller droplets at pH close to neutral, whereas soy lecithin was more efficient at lower pHs. Emulsions stabilized by rapeseed protein had higher stability during storage compared to emulsions prepared by soy lecithin. Precipitation pH during the protein extraction process had a strong impact on the emulsion stability. RPP3 stabilized emulsions with higher stability in pHs close to neutral, whereas the opposite was found for RPP6.5, which stabilized more stable emulsions in acidic conditions. Rapeseed proteins recovered from cold-pressed RPC could be a suitable natural emulsifier and precipitation pH can be used to monitor the stability in emulsions with different pHs.

Preservation of the Antioxidant Capacity of Resveratrol via Encapsulation in Niosomes.

Resveratrol (RSV) is a natural polyphenol which produces several benefits to human health, being the trans-isomer the most bioactive. However, its systemic absorption is limited due to its low water solubility, that reduces the oral bioavailability, and its chemical instability (owing to the trans-cis RSV isomer conversion upon light irradiation). Thus, encapsulation of this bioactive compound is required to protect it from destructive environmental conditions. Here, trans-RSV was encapsulated in food grade nanovesicles formed by Tween 80 and Span 80, with or without the addition of dodecanol (Dod) as membrane stabilizer. The size and shape of niosomes were evaluated by microscopy (TEM) and light scattering. RSV was successfully encapsulated in the vesicular systems (49-57%). The effect of Dod in the membrane bilayer was evaluated on the RSV in vitro release experiments under simulated gastrointestinal conditions. The total antioxidant capacity of the encapsulated polyphenol was measured using radicals' assays (DPPH and ABTS). The niosomes were able to maintain almost the total antioxidant capacity of encapsulated RSV, also preserved the ~85% of trans-RSV, thus offering considerable protection against high energy irradiation. These results make these systems suitable for different applications, particularly for photosensitive compounds.

Nanotechnology for Natural Medicine: Formulation of Neem Oil Loaded Phospholipid Vesicles Modified with Argan Oil as a Strategy to Protect the Skin from Oxidative Stress and Promote Wound Healing.

Neem oil, a plant-derived product rich in bioactives, has been incorporated in liposomes and hyalurosomes modified by adding argan oil and so called argan-liposomes and argan-hyalurosomes. Argan oil has also been added to the vesicles because of its regenerative and protective effects on skin. In the light of this, vesicles were specifically tailored to protect the skin from oxidative stress and treat lesions. Argan-liposomes were the smallest vesicles (~113 nm); the addition of sodium hyaluronate led to an increase in vesicle size (~143 nm) but it significantly improved vesicle stability during storage. In vitro studies confirmed the free radical scavenging activity of formulations, irrespective of their composition. Moreover, rheological investigation confirmed the higher viscosity of argan-hyalurosomes, which avoid formulation leakage after application. In vitro studies performed by using the most representative cells of the skin (i.e., keratinocytes and fibroblasts) underlined the ability of vesicles, especially argan-liposomes and argan-hyalurosomes, to counteract oxidative stress induced in these cells by using hydrogen peroxide and to improve the proliferation and migration of cells ensuring the more rapid and even complete closure of the wound (scratch assay).

Nano-Encapsulation of Mithramycin in Transfersomes and Polymeric Micelles for the Treatment of Sarcomas.

Sarcomas are aggressive tumors which often show a poor response to current treatments. As a promising therapeutic alternative, we focused on mithramycin (MTM), a natural antibiotic with a promising anti-tumor activity but also a relevant systemic toxicity. Therefore, the encapsulation of MTM in nano-delivery systems may represent a way to increase its therapeutic window. Here, we designed novel transfersomes and PLGA polymeric micelles by combining different membrane components (phosphatidylcholine, Span 60, Tween 20 and cholesterol) to optimize the nanoparticle size, polydispersity index (PDI) and encapsulation efficiency (EE). Using both thin film hydration and the ethanol injection methods we obtained MTM-loaded transferosomes displaying an optimal hydrodynamic diameter of 100-130 nm and EE values higher than 50%. Additionally, we used the emulsion/solvent evaporation method to synthesize polymeric micelles with a mean size of 228 nm and a narrow PDI, capable of encapsulating MTM with EE values up to 87%. These MTM nano-delivery systems mimicked the potent anti-tumor activity of free MTM, both in adherent and cancer stem cell-enriched tumorsphere cultures of myxoid liposarcoma and chondrosarcoma models. Similarly to free MTM, nanocarrier-delivered MTM efficiently inhibits the signaling mediated by the pro-oncogenic factor SP1. In summary, we provide new formulations for the efficient encapsulation of MTM which may constitute a safer delivering alternative to be explored in future clinical uses.

Microemulsion Synthesis of Superparamagnetic Nanoparticles for Bioapplications.

Superparamagnetic nanoparticles have seen increased potential in medical and environmental applications. Their preparation is traditionally made by the coprecipitation method, with limited control over the particle size distribution. Microemulsion methods could be advantageous due to the efficient control of the size, shape, and composition of the nanoparticles obtained. Water-in-oil (W/O) microemulsions consist of aqueous microdomains dispersed in a continuous oil phase, stabilized by surfactant molecules. These work as nanoreactors where the synthesis of the desired nanoparticles takes place through a co-precipitation chemical reaction. In this work, superparamagnetic magnetite nanoparticles with average diameters between 5.4 and 7.2 nm and large monodispersity have been synthesized through precipitation in a W/O microemulsion, with Cetyl Trimethyl Ammonium Bromide (CTAB) as a main surfactant, 1-butanol as a cosurfactant, and with 1-hexanol as the continuous oily phase. The optimization of the corresponding washing protocol has also been established since a strict control is required when using these materials for bioapplications. Their applicability in those has been proved by their encapsulation in liposomes, being tested as signal enhancers for lateral flow immunoassays by using the affinity neutravidin-biotin model system. Due to their magnetic behaviour, they were also tested for magnetic separation. These novel materials have been found to be useful for analytical applications requiring high sensitivity and the removal of interferences.

Electrodecoration and Characterization of Superparamagnetic Iron Oxide Nanoparticles with Bioactive Synergistic Nanocopper: Magnetic Hyperthermia-Induced Ionic Release for Anti-Biofilm Action.

The urgency for the availability of new antibacterial/disinfectant agents has become a worldwide priority. At the same time, along with the extensive use of other metal nanoparticles (NPs), the investigation of magnetic NPs (MNPs) in antibacterial studies has turned out to be an increasingly attractive research field. In this context, we present the preparation and characterization of superparamagnetic iron oxide NPs, electrodecorated with antimicrobial copper NPs, able to modulate the release of bioactive species not only by the NP's stabilizer, but also through the application of a suitable magnetic field. Antimicrobial synergistic CuNPs stabilized by benzalkonium chloride have been used in the current study. We demonstrate the successful preparation of Cu@Fe3O4 MNPs composites through morphological and spectroscopic results. Additionally, an extensive magnetic characterization is reported, along with hyperthermia-induced copper ionic release. On the basis of our results, we propose a new generation of antimicrobial magnetic nanomaterials, whose bioactivity can be also tuned by the application of a magnetic field.

Addition of Trans-Resveratrol-Loaded Highly Concentrated Double Emulsion to Yoghurts: Effect on Physicochemical Properties.

Trans-resveratrol (RSV) needs to be encapsulated to maintain its beneficial properties on the human body. This is due to its extreme photosensitivity, short biological half-life, and easy oxidation. In this study, the use of double emulsions for RSV encapsulation and their further application on functional yoghurts was studied. Different types of yoghurts were prepared: with and without RSV and with two types of volumetric emulsion formulations (20/80 and 30/70). In order to study the influence of the addition of double emulsions to the physical properties of the prepared yoghurts, they were characterised fresh and after a month under storage at 4 °C, in terms of droplet size, morphology, stability, rheology, texturometry, colorimetry, and antioxidant capacity. Results obtained showed that the presence of emulsion in the yoghurts produced a generalised decrease in the predominant droplet size (from 48 µm to 15-25 µm) and an increase in the stability. Additionally, a predominantly elastic character was observed. The firmness values obtained were very similar for all the yoghurts analysed and did not suffer important modifications with time. A slight colour variation was observed with storage time in the control sample, whereas a more notable variation in the case of emulsion yoghurts was observed. An appreciable increase of the antioxidant capacity of the final functional yoghurt (100 g) was observed when it contained 5-8 mg of RSV. Encapsulated RSV added to yoghurts presented a larger protection against RSV oxidation compared with free RSV, presenting a larger antioxidant inhibition after one month of storage. Moreover, the antioxidant capacity of yoghurts with encapsulated RSV was not affected under storage, since slight reductions (3%) were registered after one month of storage at 4 °C.

Simultaneous encapsulation of trans-resveratrol and vitamin D3 in highly concentrated double emulsions.

BACKGROUND: Encapsulation of biocompounds is essential to protect them from environmental factors that could enhance their oxidation or cause them to lose their beneficial properties due to extreme photosensitivity, among other factors. The main goal of this work was to study the feasibility of preparing concentrated double emulsions with a high loading capacity containing simultaneously trans-resveratrol (RSV) and vitamin D3 (VitD3 ). Such emulsions could be used for food fortification or pharmaceutical formulations or as vehicles for targeted controlled release. RESULTS: In order to achieve large concentrations of the encapsulated compounds, all the double emulsions were formulated using a W1 /O in W2 ratio of 80/20, while the ratios tested for W1 in O where 20/80 and 30/70. All the emulsions were characterized by droplet size, morphology, colloidal stability and encapsulation efficiency (EE) over a period of 6 weeks. VitD3 and RSV concentration were determined by a technique based on reverse-phase high-performance liquid chromatography. The viability of preparing concentrated W1 /O/W2 emulsions containing both biocompounds has been demonstrated with satisfactory results. Initial RSV concentrations in the concentrated double emulsions formulated varied from 5.0 to 8.3 mg L-1 , while for VitD3 values of 28-32 mg L-1 were obtained. The presence of VitD3 retarded RSV release in the formulated emulsions. It was observed that after 1 week of storage RSV EE increased around 10-50% when VitD3 was simultaneously encapsulated. CONCLUSION: Simultaneous encapsulation of RSV and VitD3 was possible in high internal phase emulsions. The emulsions presented high colloidal stability, being suitable for food fortification applications. © 2020 Society of Chemical Industry.

Vitamin D3 Loaded Niosomes and Transfersomes Produced by Ethanol Injection Method: Identification of the Critical Preparation Step for Size Control.

Vesicular nanocarriers have an important role in drug delivery and dietary supplements. Size control and optimization of encapsulation efficiency (EE) should be optimized for those applications. In this work, we report on the identification of the crucial step (injection, evaporation, or sonication) innanovesicles (transfersomes and niosomes) preparation by theethanol injection method (EI). The identification of each production step on the final vesicle size was analyzed in order to optimize further scale-up process. Results indicated that the final size of transfersomeswas clearly influenced by the sonication step while the final size of niosomes was mainly governed by the injection step. Measurements of final surface tension of the different vesicular systems prepared indicate a linear positive tendency with the vesicle size formed. This relation could help to better understand the process and design a vesicular size prediction model for EI. Vitamin D3 (VitD3) was encapsulated in the systems formulated with encapsulation efficiencies larger than 90%. Interaction between the encapsulated compound and the membrane layer components is crucial for vesicle stability. This work has an impact on the scaling-up production of vesicles for further food science applications.