Nintedanib and miR-29b co-loaded lipoplexes in idiopathic pulmonary fibrosis: formulation, characterization, and in vitro evaluation.

OBJECTIVE: This study was aimed to develop a cationic lipoplex formulation loaded with Nintedanib and miR-29b (LP-NIN-miR) as an alternative approach in the combination therapy of idiopathic pulmonary dibrosis (IPF) by proving its additive anti-fibrotic therapeutic effects through in vitro lung fibrosis model. SIGNIFICANCE: This is the first research article reported that the LP-NIN-MIR formulations in the treatment of IPF. METHODS: To optimize cationic liposomes (LPs), quality by design (QbD) approach was carried out. Optimized blank LP formulation was prepared with DOTAP, CHOL, DOPE, and DSPE-mPEG 2000 at the molar ratio of 10:10:1:1. Nintedanib loaded LP (LPs-NIN) were produced by microfluidization method and were incubated with miR-29b at room temperature for 30 min to obtain LP-NIN-miR. To evaluate the cellular uptake of LP-NIN-miR, NIH/3T3 cells were treated with 20 ng.mL-1 transforming growth factor-ß1 (TGF-ß1) for 96 h to establish the in vitro IPF model and incubated with LP-NIN-miR for 48 h. RESULTS: The hydrodynamic diameter, polydispersity index (PDI), and zeta potential of the LP-NIN-miR were 87.3 ± 0.9 nm, 0.184 ± 0.003, and +24 ± 1 mV, respectively. The encapsulation efficiencies of Nintedanib and miR-29b were 99.8% ± 0.08% and 99.7% ± 1.2%, respectively. The results of the cytotoxicity study conducted with NIH/3T3 cells indicated that LP-NIN-miR is a safe delivery system. CONCLUSIONS: The outcome of the transfection study proved the additive anti-fibrotic therapeutic effect of LP-NIN-miR and suggested that lipoplexes are effective delivery systems for drug and nucleic acid to the NIH/3T3 cells in the treatment of IPF.

Microfluidization improved hempseed yogurt's physicochemical and storage properties.

BACKGROUND: Plant-based yogurts are suffering from the common problems, such as an unattractive color, stratified texture state and rough taste. Therefore, it is urgent to develop a novel processing method to improve the quality and extend the storage life of hempseed yogurt. In the present study, hempseed yogurt was microfluidized prior to fermentation. The effects of microfluidization on microstructure, particle size, mechanical properties, sensory acceptability, variations in pH and titratable acidity, lactic acid bacteria (LAB) counts, and stability of hempseed yogurt during 20 days of storage were investigated. RESULTS: Microfluidization contributed to the production of hempseed yogurt as a result of the better physicochemical properties compared to normal homogenization. Specifically, microfluidization reduced the particle size of hempseed yogurt with a uniform particle distribution, increased water holding capacity, and improved texture and rheological properties. These advancements resulted in higher sensory scores for the yogurt. Furthermore, during storage, microfluidization effectively inhibited the post-acidification process of hempseed yogurt, and increased LAB counts and storage stability. CONCLUSION: Microfluidization improved the physicochemical properties and storage stability of hempseed yogurt. Our findings support the application of microfluidization in hempseed yogurt and provide a new approach for enhancing the quality of plant-based alternatives that meet consumers' demands for high-quality food products. © 2023 Society of Chemical Industry.

Microfluidized hemp protein isolate: an effective stabilizer for high-internal-phase emulsions with improved oxidative stability.

BACKGROUND: Hemp protein isolates (HPIs), which provide a well-balanced profile of essential amino acids comparable to other high-quality proteins, have recently garnered significant attention. However, the underutilized functional attributes of HPIs have constrained their potential commercial applications within the food and agriculture field. This study advocates the utilization of dynamic-high-pressure-microfluidization (DHPM) for the production of stable high-internal-phase emulsions (HIPEs), offering an efficient approach to fully exploit the potential of HPI resources. RESULTS: The findings underscore the effectiveness of DHPM in producing HPI as a stabilizing agent for HIPEs with augmented antioxidant activity. Microfluidized HPI exhibited consistent adsorption and anchoring at the oil-water interface, resulting in the formation of a dense and compact layer. Concurrently, the compression of droplets within HIPEs gave rise to a polyhedral framework, conferring viscoelastic properties and a quasi-solid behavior to the emulsion. Remarkably, HIPEs stabilized by microfluidized HPI demonstrated superior oxidative and storage stability, attributable to the establishment of an antioxidative barrier by microfluidized HPI particles. CONCLUSION: This study presents an appealing approach for transforming liquid oils into solid-like fats using HPI particles, all without the need for surfactants. HIPEs stabilized by microfluidized HPI particles hold promise as emerging food ingredients for the development of emulsion-based formulations with enhanced oxidative stability, thereby finding application in the food and agricultural industries. © 2023 Society of Chemical Industry.

Microfluidizing Technique Application for Algerian Cymbopogon citratus (DC.) Stapf Effects Enhanced Volatile Content, Antimicrobial, and Anti-Mycotoxigenic Properties.

Medicinal plant extracts are a promising source of bioactive minor contents. The present study aimed to evaluate the distinguished volatile content of Algerian Cymbopogon citratus (DC.) Stapf before and after the microfluidization process and their related antimicrobial and anti-mycotoxigenic impacts and changes. The GC-MS apparatus was utilized for a comparative examination of Algerian lemongrass essential oil (LGEO) with its microfluidization nanoemulsion (MF-LGEO) volatile content. The MF-LGEO was characterized using Zetasizer and an electron microscope. Cytotoxicity, antibacterial, and antifungal activities were determined for the LGEO and MF-LGEO. The result reflected changes in the content of volatiles for the MF-LGEO. The microfluidizing process enhanced the presence of compounds known for their exceptional antifungal and antibacterial properties in MF-LGEO, namely, neral, geranial, and carvacrol. However, certain terpenes, such as camphor and citronellal, were absent, while decanal, not found in the raw LGEO, was detected. The droplet diameter was 20.76 ± 0.36 nm, and the polydispersity index (PDI) was 0.179 ± 0.03. In cytotoxicity studies, LGEO showed higher activity against the HepG2 cell line than MF-LGEO. Antibacterial LGEO activity against Gram-positive bacteria recorded an inhibitory zone from 41.82 ± 2.84 mm to 58.74 ± 2.64 mm, while the zone ranged from 12.71 ± 1.38 mm to 16.54 ± 1.42 mm for Gram-negative bacteria. Antibacterial activity was enhanced to be up to 71.43 ± 2.54 nm and 31.54 ± 1.01 nm for MF-LGEO impact against Gram-positive and Gram-negative pathogens. The antifungal effect was considerable, particularly against Fusarium fungi. It reached 17.56 ± 1.01 mm and 13.04 ± 1.37 mm for LGEO and MF-LGEO application of a well-diffusion assay, respectively. The MF-LGEO was more promising in reducing mycotoxin production in simulated fungal growth media due to the changes linked to essential compounds content. The reduction ratio was 54.3% and 74.57% for total aflatoxins (AFs) and ochratoxin A (OCA) contents, respectively. These results reflect the microfluidizing improvement impact regarding the LGEO antibacterial, antifungal and anti-mycotoxigenic properties.

Effects of Microfluidization on the Physical and Storage Stability of Walnut Protein Emulsion and Beverages.

Walnut meal is a by-product produced during the production of walnut oil and is often treated as a waste. However, the nutrients in walnut meal mean it has significant potential for development as a plant-based milk. This study investigated the effect of microfluidization on the stability of walnut protein emulsion (WPE) and walnut protein beverage (WPB) produced from walnut meal, compared with conventional homogenization. The particle size, zeta potential, rheological properties, and stability of WPE all significantly improved after microfluidization. The mean particle size and zeta potential of the microfluidized WPE significantly decreased (p < 0.05). The rheological properties demonstrated that the viscosity of the microfluidized WPE decreased by 80%, and that the shear force increased 4.5 times as the shear rate increased. This gave the resulting product the characteristics of non-Newtonian fluid. LUMisizer stability demonstrated that microfluidization improves stability through protein absorption on the oil-water interface. Microfluidization increased the denaturation temperature (Tm) of WPE from 135.65 to 154.87℃. Moreover, microfluidization improved the color, centrifugal precipitation rate, and viscosity in WPB compared to the control at all studied temperatures. The Arrhenius approach was used to establish a shelf-life model, which predicted that microfluidized WPB could be stored for 175 d at 4℃. This study provided a new reference for the widespread application of microfluidization in the production of food-based emulsion and beverage products.

Latest developments in the applications of microfluidization to modify the structure of macromolecules leading to improved physicochemical and functional properties.

Microfluidization is a unique high-pressure homogenization technique combining various forces such as high-velocity impact, high-frequency vibration, instantaneous pressure drop, intense shear rate, and hydrodynamic cavitation. Even though it is mainly used on emulsion-based systems and known for its effects on particle size and surface area, it also significantly alters physicochemical and functional properties of macromolecules including hydration properties, solubility, viscosity, cation-exchange capacity, rheological properties, and bioavailability. Besides, the transformation of structure and conformation due to the combined effects of microfluidization modifies the material characteristics that can be a base for new innovative food formulations. Therefore, microfluidization is being commonly used in the food industry for various purposes including the formation of micro- and nano-sized emulsions, encapsulation of easily degradable bioactive compounds, and improvement in functional properties of proteins, polysaccharides, and dietary fibers. Although the extent of modification through microfluidization depends on processing conditions (e.g., pressure, number of passes, solvent), the nature of the material to be processed also changes the outcomes significantly. Therefore, it is important to understand the effects of microfluidization on each food component. Overall, this review paper provides an overview of microfluidization treatment, summarizes the applications on macromolecules with specific examples, and presents the existing problems.

Dynamic high pressure treatments: current advances on mechanistic-cum-transport phenomena approaches and plant protein functionalization.

Dynamic high pressure treatment (DHPT) either by high pressure homogenization or microfluidisation, is an emerging concept used in the food industry for new products development through macromolecules modifications in addition to simple mixing and emulsification action. Mechanistic understanding of droplets breakup during high pressure homogenization is used to understand how these compact and high molecular weight-sized globular plant proteins are affected during DHPTs. Plant protein needs to be functionalized for advanced use in food formulation. DHPTs brought changes in plant proteins' secondary, tertiary, and quaternary structures through alterations in intermolecular and intramolecular interactions, sulfhydryl groups, and disulfide bonds. These structural changes in plant proteins affected their functional and physicochemical properties like solubility, oil and water holding capacity, gelation, emulsification, foaming, and rheological properties. These remarkable changes made utilization of this concept in novel food system applications like in plant-based dairy analogues. Overall, this review provides a comprehensive and critical understanding of DHPTs on their mechanistic and transport approaches for droplet breakup, structural and functional modification of plant macromolecules. This article also explores the potential of DHPT for formulating plant-based dairy analogues to meet healthy and sustainable food consumption needs. HIGHLIGHTSIt critically reviews high pressure homogenization (HPH) and microfluidisation (DHPM).It explores the mechanistic and transport phenomena approaches of HPH and DHPMHPH and DHPM can induce conformational and structural changes in plant proteins.Improvement in the functional properties of HPH and DHPM treated plant proteins.HPH and DHPM are potentially applicable for plant based dairy alternatives food system.

Elaboration and characterization of nanoemulsion with orange essential oil and pectin.

BACKGROUND: Nanoemulsions formulated with citric essential oils are currently of interest because of their physical and chemical properties and multiple applications in areas such as the food industry or agrochemicals. These are thermodynamically unstable and have almost Newtonian flow behaviour, but a suitable formulation allows systems to be obtained with good physical stability and rheological properties. The addition of pectin makes this possible. In this work, food nanoemulsions formulated with pectin, orange essential oil (5 wt%), and Tween 80 were obtained by microfluidization. First, the effect of Tween 80 concentration from 1 to 5 wt% on emulsions without pectin was evaluated. Then, pectin was added to the most stable nanoemulsion obtained and two variables were studied: the pectin solution concentration (from 2 to 6 wt%) and the pectin/emulsion ratio (1:1 or 2:1) at a fixed pectin concentration. RESULTS: Rheological, laser diffraction, and multiple light scattering techniques were employed to determine the content of Tween 80 that results in the most stable nanoemulsion without pectin, which was 3 wt%. In addition, these techniques were used to determine the structure and physical stability of the nanoemulsions containing orange essential oil and pectin. The results obtained showed that the emulsions containing 2 wt% pectin were destabilized before 24 h. Furthermore, the emulsion with 6 wt% pectin and a 2:1 pectin/emulsion ratio showed the highest viscosity and the lowest mean diameters, and therefore the greatest stability. CONCLUSION: This work extends the knowledge of formulation of nanoemulsions and using essential oils. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Usability of microfluidized flaxseed as a functional additive in bread.

BACKGROUND: Flaxseed is a rich source of protein, omega-3 fatty acids, lignans, and dietary fiber. However, it also contains phytic acid, which inhibits mineral absorption and has the potential to adversely affect the properties of bread. Microfluidization prevents these negative effects, reduces the amount of phytic acid, and improves functional properties. In this study, the possibility of using full-fat and defatted flaxseed flours as well as microfluidized flaxseed flours in bread formulation was investigated. For this purpose, crude and microfluidized flaxseed flours were added to the bread in different proportions (0, 25, 50, and 75 g kg-1 ), and the effects of the partial replacement of wheat flour with flaxseed flours on the functional, quality, and sensory properties of breads were analyzed. The effects of the microfluidization process on the antioxidant properties, phenolic, dietary fiber, and phytic acid content of flaxseed were also observed. RESULT: Flaxseed flours increased the dietary fiber, phenolic contents, and antioxidant activities of breads. The crumb color became darker with increasing level of flaxseed flours, and their addition also detrimentally affected the sensory properties of breads. It was seen that the microfluidization process has beneficial effects on functional properties of full-fat and defatted flaxseed flours, as well as on their quality characteristics. CONCLUSION: The study showed that flaxseed flour is a rich source of functional compounds, and it is even possible to further improve these functional properties with microfluidization treatment. Microfluidized flaxseed flour can also be used as a promising alternative functional food to enrich breads. © 2021 Society of Chemical Industry.

Ritonavir nanosuspensions prepared by microfluidization with enhanced solubility and desirable immunological properties.

The objective of this study was to develop ritonavir (RTV) nanosuspensions (NSs) by microfluidization method. Particle size (PS) measurements were performed by photon correlation spectroscopy. Amorphous properties of the particles were evaluated by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The dissolution studies were conducted in fed state simulated intestinal fluid (FeSSIF) medium. The flow cytometry was utilized to determine the lymphocyte sub-groups and immune response of NSs. RTV NSs were obtained with 400-500 nm PS. The crystal properties of RTV remain unchanged. The solubility of NS was enhanced five times. 57% and 18% of RTV were dissolved in FeSSIF medium for NSs and coarse powder. According to immunological studies, the prepared NSs did not significantly alter the ratio of CD4+/CD8+. Therefore, NSs may be a beneficial approach for the oral administration of RTV.

Investigating Antiarthritic Potential of Nanostructured Clove Oil (Syzygium aromaticum) in FCA-Induced Arthritic Rats: Pharmaceutical Action and Delivery Strategies.

The combined application of clove oil in a lipid nanocarrier opens a promising avenue for bone and joints therapy. In this study, we successfully developed a tunable controlled-release lipid platform for the efficient delivery of clove oil (CO) for the treatment of rheumatoid arthritis (RA). The ultra-small nanostructured lipid carriers co-loaded with CO (CONCs) were developed through an aqueous titration method followed by microfluidization. The CONCs appeared to be spherical (particle size of 120 nm), stable (zeta potential of -27 mV), and entrapped efficiently (84.5%). In toluene:acetone:glacial acetic acid (90:9:1 percent v/v/v) solvent systems, high-performance thin layer chromatography (HPTLC) analysis revealed the primary components in CO as eugenol (RF = 0.58). The CONCs greatly increased the therapeutic impact of CO in both in vitro and in vivo biological tests, which was further supported by excellent antiarthritic action. The CONC had an antiarthritic activity that was slightly higher than neat CO and slightly lower than standard, according to our data. The improved formulation inhibited serum lysosomal enzymes and proinflammatory cytokines while also improving hind leg function. This study provides a proof of concept to treat RA with a new strategy utilizing essential oils via nanodelivery.

The applications of microfluidization in cereals and cereal-based products: An overview.

Although, the consumption of food consisting of fiber presents some important nutritional, functional and health benefits, manufacturers and researchers have reported that the use of high amount of fiber worsens the product quality. Besides, consuming large quantities of dietary fiber delays intestinal gas transit. In cereal grains, phenolic compounds are covalently bound to indigestible polysaccharides thus this complex bran matrix restricts its release in small intestine resulting in low bioavailability. Therefore, in order to overcome the problems related to the characteristics of fiber, the use of large quantities of dietary fiber in cereal based products as well as the low bioavailability of phenolic compounds; food scientists explore alternative milling methods to traditional treatments. The potential use of microfluidization in cereal-based products including wheat bran, corn bran, zein, rice bran and starches has been highlighted. Functionalization through microfluidization has been applied as a prospective method for production of fibrous structures from cereal brans and it improves surface areas, water holding capacity, swelling capacity, porosity, oil-holding capacity, cation-exchange capacity and the exposure of the phenolic compounds and hence the associated antioxidant capacity of fibers. Microfluidization also offers a promising method for the formation of complexes between starches and a fatty acid, which has potential to create a new functional resistant starch ingredient with increased viscosity and improved water-holding properties. Microfluidized cereal by-products provide some important unique functional and nutritional properties to bakery products. In this perspective, this paper provides an overview of the findings on the use of microfluidization in cereals and cereal-based products.

Characteristics, Cryoprotection Evaluation and In Vitro Release of BSA-Loaded Chitosan Nanoparticles.

Chitosan nanoparticles (CS-NPs) are under increasing investigation for the delivery of therapeutic proteins, such as vaccines, interferons, and biologics. A large number of studies have been taken on the characteristics of CS-NPs, and very few of these studies have focused on the microstructure of protein-loaded NPs. In this study, we prepared the CS-NPs by an ionic gelation method, and bovine serum albumin (BSA) was used as a model protein. Dynamic high pressure microfluidization (DHPM) was utilized to post-treat the nanoparticles so as to improve the uniformity, repeatability and controllability. The BSA-loaded NPs were then characterized for particle size, Zeta potential, morphology, encapsulation efficiency (EE), loading capacity (LC), and subsequent release kinetics. To improve the long-term stability of NPs, trehalose, glucose, sucrose, and mannitol were selected respectively to investigate the performance as a cryoprotectant. Furthermore, trehalose was used to obtain re-dispersible lyophilized NPs that can significantly reduce the dosage of cryoprotectants. Multiple spectroscopic techniques were used to characterize BSA-loaded NPs, in order to explain the release process of the NPs in vitro. The experimental results indicated that CS and Tripolyphosphate pentasodium (TPP) spontaneously formed the basic skeleton of the NPs through electrostatic interactions. BSA was incorporated in the basic skeleton, adsorbed on the surface of the NPs (some of which were inlaid on the NPs), without any change in structure and function. The release profiles of the NPs showed high consistency with the multispectral results.

Effect of high-pressure microfluidization on nutritional quality of carrot (Daucus carota L.) juice.

In this study, the effect of high-pressure microfluidization on the colour and nutritional qualities of the orange carrot juice was investigated. The juice was processed at three different pressures (34.47 MPa, 68.95 MPa and 103.42 MPa) with three different passes (1, 2 and 3 passes). After that, total phenolic content (TPC), antioxidant activity, carotenoids, color properties, and total soluble solids content of the processed carrot juice were evaluated. As a result, no specific trends in TPC and antioxidant activity of the juice were observed through the variations of processing conditions. However, microfluidization significantly (p < 0.05) improved the carotenoids content in carrot juice. With increasing number of pass, concentrations of ß-carotene and lutein had increased significantly. Similarly, increasing process pressure initially increased carotenoid content significantly (up to 68.95 MPa), further increase pressure to 103.42 MPa did not cause significant changes in carotenoid concentration. Furthermore, color properties such as lightness, redness, yellowness, and chroma value were reduced significantly with the increase of pressure and the number of passes. The results indicated that high-pressure microfluidization could be used as a novel alternative nonthermal technology to heat pasteurization to improve the color and nutritional qualities in orange carrot juice, resulting in a desirable, high-quality juice for consumers.

Short communication: Volatiles in microfluidized raw and heat-treated milk.

As innovative processing equipment is introduced to milk processing, it is essential to determine its effect on milk aroma, a critical factor in consumer acceptance of the final dairy product. Microfluidization is known to cause severe high-pressure homogenization of milk fat and, although severe processing is known to release undesired aromas, no information is available on the levels of the volatile compounds in milk immediately after microfluidization. We hypothesized that microfluidization would alter levels of volatile compounds in milk that may affect aroma. The concentration of 11 selected volatile compounds in raw, thermized, pasteurized, and UHT 3.0% fat milk samples were compared before and after microfluidization at 170 MPa and common 2-stage homogenization at 15 MPa. Overall, the different milk samples had similar trends in response to homogenization, although UHT milk started with lower values of nonanoic acid, and acetone and higher levels of hexanal and heptanol. In many cases, microfluidization did not significantly alter volatile levels compared with the starting milk. Heptanal was the only compound observed to increase in thermized and UHT milk, whereas nonanoic acid and acetone decreased in raw, thermized, and pasteurized milks and octanoic acid decreased in thermized and UHT milks. The highest levels of almost all of the volatiles were found in the 2-stage homogenized milk. Overall, microfluidization had minimal effect on the volatile compound profiles of milk, although sensory evaluation is needed to confirm effects on aroma and flavor.

Preparation of Nanoemulsions of Vitamin A and C by Microfluidization: Efficacy on the Expression Pattern of Milk-Specific Proteins in MAC-T Cells.

In this study, we prepared stabilized vitamin A and C nanoemulsions, and investigated their efficacy on milk-specific proteins in bovine mammary epithelial cells (MAC-T). Emulsions of vitamin A (vit-A) and C (vit-C) were prepared using Lipoid S 75 and microfluidization. The particle size and polydispersity index (PDI) of nanoemulsified vit-A and vit-C were studied. The cytotoxic effect of nanoemulsion-free and nanoemulsified vit-A and vit-C was determined by an MTT assay. In addition, the efficacy of nanoemulsified vit-A and vit-C on the in vitro expression pattern of milk-specific proteins in MAC-T cells was investigated by quantitative RT-PCR. The results showed that the efficacies of stabilized nanoemulsions of vit-A and vit-C were 100% and 92.7%, respectively. The particle sizes were around 475.7 and 225.4 nm, and the zeta potentials were around -33.5 and -21.3 mV, respectively. The expression changes of αs2-, ß- and κ-casein were higher in the presence of a stabilized nanoemulsion of vit-A, compared with nanoemulsion-free vit-A. Furthermore, the expression changes of αs2- and ß-casein were lower and that of κ-casein was higher in the presence of a stabilized nanoemulsion of vit-C, compared with nanoemulsion-free vit-C. Thus, our findings demonstrate the efficacy of nanoemulsified vit-A and vit-C in changing the expression of milk-specific proteins in MAC-T cells.

Comparison of the effects of dynamic high-pressure microfluidization and conventional homogenization on the quality of peach juice.

BACKGROUND: Dynamic high-pressure microfluidization (DHPM) is an emerging and promising technique for continuous production of fluid foods. This study aimed to investigate the influence of DHPM and conventional homogenization (CH) on the quality of peach juice. Processing was performed by passing peach juice through CH at 20 MPa and DHPM at 20-160 MPa for one or three passes. The effect of DHPM pressure and passing number were also assessed. RESULTS: The results indicate that DHPM could maintain the antioxidant activity of peach juice much better than CH processing. Total phenolic compounds were decreased by 11.7% and 7.9%-15.8% through CH and DHPM processing in different conditions. Moreover, particle size, non-enzymatic browning index and turbidity decreased significantly under DHPM and CH processing, and decreased more and more with the increasing of DHPM pressure and treatment times. However, vitamin C content and zeta-potential did not reveal remarkable variation before and after these two types of processing. CONCLUSION: Taken together, DHPM is able to maintain the quality and stability of peach juice, which can be a reliable technological alternative to CH to produce fresh-like peach juices. © 2019 Society of Chemical Industry.

The effects of microfluidization on the physical, microbial, chemical, and coagulation properties of milk.

This work examines the use of mild heat treatments in conjunction with 2-pass microfluidization to generate cheese milk for potential use in soft cheeses, such as Queso Fresco. Raw, thermized, and high temperature, short time pasteurized milk samples, standardized to the 3% (wt/wt) fat content used in cheesemaking, were processed at 4 inlet temperature and pressure conditions: 42°C/75 MPa, 42°C/125 MPa, 54°C/125 MPa, and 54°C/170 MPa. Processing-induced changes in the physical, chemical, and microbial properties resulting from the intense pressure, shear, and cavitation that milk experiences as it is microfluidized were compared with nonmicrofluidized controls. A pressure-dependent increase in exit temperature was observed for all microfluidized samples, with inactivation of alkaline phosphatase in raw and thermized samples at 125 and 170 MPa. Microfluidization of all samples under the 4 inlet temperature and processing pressure conditions resulted in a stable emulsion of fat droplets ranging from 0.390 to 0.501 µm, compared with 7.921 (control) and 4.127 (homogenized control) µm. Confocal imaging showed coalescence of scattered fat agglomerates 1 to 3 µm in size during the first 24 h. We found no changes in fat, lactose, ash content or pH, indicating the major components of milk remained unaffected by microfluidization. However, the apparent protein content was reduced from 3.1 to 2.2%, likely a result of near infrared spectroscopy improperly identifying the micellar fragments embedded into the fat droplets. Microbiology results indicated a decrease in mesophilic aerobic and psychrophilic milk microflora with increasing temperature and pressure, suggesting that microfluidization may eliminate bacteria. The viscosities of milk samples were similar but tended to be higher after treatment at 54°C and 125 or 170 MPa. These samples exhibited the longest coagulation times and the weakest gel firmness, indicating that formation of the casein matrix, a critical step in the production of cheese, was affected. Low temperature and pressure (42°C/75 MPa) exhibited similar coagulation properties to controls. The results suggest that microfluidization at lower pressures may be used to manufacture high-moisture cheese with altered texture whereas higher pressures may result in novel dairy ingredients.

Coencapsulation of Polyphenols and Anthocyanins from Blueberry Pomace by Double Emulsion Stabilized by Whey Proteins: Effect of Homogenization Parameters.

Blueberry pomace is a rich source of high-value bioactive polyphenols with presumed health benefits. Their incorporation into functional foods and health-related products benefits from coencapsulation and protection of polyphenol-rich extracts in suitable carriers. This study aimed to create a water-in-oil-in-water (W1/O/W2) double emulsion system suitable for the coencapsulation of total phenolics (TP) and anthocyanins (TA) from a polyphenol-rich extract of blueberry pomace (W1). The effect of critical physical parameters for preparing stable double emulsions, namely homogenization pressure, stirring speed and time, was investigated by measuring the hydrodynamic diameter, size dispersity and zeta potential of the oil droplets, and the encapsulation efficiency of TP and TA. The oil droplets were negatively charged (negative zeta potential values), which was related to the pH and composition of W2 (whey protein isolate solution) and suggests stabilization by the charged whey proteins. Increasing W1/O/W2 microfluidization pressure from 50 to 200 MPa or homogenization speed from 6000 to 12,000 rpm significantly increased droplet diameter and zeta potential and decreased TA and TP encapsulation efficiency. Increasing W1/O/W2 homogenization time from 15 to 20 min also increased droplet diameter and zeta potential and lowered TA encapsulation efficiency, while TP encapsulation did not vary significantly. In contrast, increasing W1/O homogenization time from 5 to 10 min at 10,000 rpm markedly increased TA encapsulation efficiency and reduced droplet diameter and zeta potential. High coencapsulation rates of blueberry polyphenols and anthocyanins around 80% or greater were achieved when the oil droplets were relatively small (mean diameter < 400 nm), with low dispersity (<0.25) and a high negative surface charge (-40 mV or less). These characteristics were obtained by homogenizing for 10 min at 10,000 rpm (W1/O), then 6000 rpm for 15 min, followed by microfluidization at 50 MPa.

Development of Biopolymer Composite Films Using a Microfluidization Technique for Carboxymethylcellulose and Apple Skin Particles.

Biopolymer films based on apple skin powder (ASP) and carboxymethylcellulose (CMC) were developed with the addition of apple skin extract (ASE) and tartaric acid (TA). ASP/CMC composite films were prepared by mixing CMC with ASP solution using a microfluidization technique to reduce particle size. Then, various concentrations of ASE and TA were incorporated into the film solution as an antioxidant and an antimicrobial agent, respectively. Fourier transform infrared (FTIR), optical, mechanical, water barrier, and solubility properties of the developed films were then evaluated to determine the effects of ASE and TA on physicochemical properties. The films were also analyzed for antioxidant effect on 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and antimicrobial activities against Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica, and Shigella flexneri. From the results, the ASP/CMC film containing ASE and TA was revealed to enhance the mechanical, water barrier, and solubility properties. Moreover, it showed the additional antioxidant and antimicrobial properties for application as an active packaging film.