Milk Fat Globule Membrane Phospholipids Research Articles

Milk fat globule membrane (MFGM) phospholipid - Whey protein interaction characterization and its effect on physicochemical, interfacial properties and evaluation of in vitro digestion of emulsions - Inspired by the MFGM

The interfacial composition of fat globules can change the interfacial structure and further affect the digestive characteristics. In this study, the interaction between whey protein (WP), which is the main protein added in infant formula (IF), and milk fat globule membrane phospholipid (MPL) was investigated from the perspective of simulated fat globule structure, we further characterized the physicochemical properties and the in vitro digestive performance of the emulsions. The results showed that the interaction between MPL and WP changed the secondary and tertiary structures of WP, and formed a dense protective layer at the interface of the fat globule with strong interaction, which improved the emulsification property and physical stability of the emulsion. When MPL: WP was 5:10, the emulsions showed smaller particle size, higher absolute value of zeta-potential and more uniform and stable emulsion state. In addition, during the simulated in vitro digestion of infants, the degree of lipolysis and free fatty acid release content were significantly increased, which improved the fat digestion.

180 Dietary milk fat globule membrane alters intestinal microbiota of neonatal piglet following lipopolysaccharide challenge and marker of neuroinflammation

Abstract Early life stress exposure can increase morbidity and reduce animal wellbeing throughout life. Dysregulated gut function has been associated with neurological changes, and neurological changes affect gut homeostasis. Maternal milk is the gold-standard for yielding developmental benefits to offspring. Identification of milk components has led to an understanding of the bioactive nutrients important in neonatal development. Components of the milk fat globule membrane (MFGM) contribute immensely to neonatal gut health and neurodevelopment. However, mechanisms of actions are still being defined. The objective is to understand how dietary MFGM modulates the gut microbiota, blood-brain barrier tight junctions, and brain inflammatory markers following lipopolysaccharide (LPS) challenge. Piglets (n = 24, one-d-old) were assigned to soy (CON) or MFGM phospholipid supplemented diet (0.75% wt/wt) ± LPS (100 µg/kg body weight) in a 2 × 2 factorial design (n = 6 animals•diet-1•challenge-1.). Piglets were fed for 21 d and 6 animals/diet received saline or LPS injections 4 h before euthanasia. Colonic microbiota was characterized through 16s rRNA gene sequencing, while hypothalamus samples were collected to evaluate brain barrier function and inflammation-related genes using RT-qPCR. Microbiota data were processed using QIIME2 with DADA2 plugin and analyzed for alpha-diversity, beta-diversity, and differential abundance in R software. Gene transcription data were analyzed by a 2 × 2 experimental factorial design using the PROC MIXED procedure of SAS. Principal coordinates analysis indicated that MFGM-fed groups exhibited an observable separation from CON-fed groups in terms of microbial community structures (P < 0.05). Firmicutes_A, Bacteroidota, and Firmicutes_D were the predominant phyla across all treatments, with average proportions to be 52.21 %, 24.36 %, and 18.46 %, respectively. At genus level, MFGM + Saline group down-regulated Corynebacterium by 88.00 % (q < 0.05) and Unclassified Muribaculaceae by 73.33 % (0.050.05). However, there was an MFGM × LPS interaction (P < 0.05) observed for nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor alpha, in which MFGM attenuated the LPS-induced increase in mRNA relative abundance (3.62 and 2.39 ± 0.38). In conclusion, dietary MFGM altered neonatal piglet intestinal microbiota and an important signaling molecule of the neuroinflammatory response following LPS challenge.

PSVI-13 Dietary Milk Fat Globule Membrane Improves Neonatal Piglet Intestinal Architecture and Enhances Mucosal Diamine Oxidase Activity Following Lipopolysaccharide Challenge

Abstract Early life stressors alter the trajectory of gut development and impair nutrient utilization, barrier function, and animal performance throughout life. Gastrointestinal (GI) maladies increase morbidity and mortality in neonates, and dietary nutrients are essential in supporting GI function. Although maternal milk is the gold-standard for yielding developmental benefits to offspring, genetic selection in the modern sow has resulted in large litter size and milk yield potentially insufficient to achieve the high biological growth potential in sow-reared piglets. Research into milk quality has led to the identification of many bioactive nutrients important in neonatal development. Components of the milk fat globule membrane (MFGM) contribute immensely to neonatal nutrition and gut health. However, mechanisms of actions are still being defined. The objective is to define the effect of MFGM supplementation on neonatal piglet intestinal health. We hypothesize dietary MFGM will enhance piglet intestinal architecture and tight junction (TJ) protein abundance, attenuating systemic lipopolysaccharide (LPS)-induced intestinal barrier disruption. One-day-old piglets (n = 32) were assigned to soy (CON) or MFGM phospholipid supplemented diet (0.75% wt/wt) ± LPS (100 µg/kg body weight) challenge in a 2 × 2 factorial design (n = 8/diet/challenge). Piglets were fed for 21 d and 8 animals/diet received saline or LPS injections 4 h before euthanasia. Piglet weights and feed intake were recorded daily to calculate performance. Intestinal tissues were collected to assess villus height (VH) and crypt depth (CD) by hematoxylin and eosin staining. Two biomarkers of intestinal integrity, intestinal fatty acid-binding protein (I-FABP) and diamine oxidase (DAO), were measured in serum or tissue. Protein abundance of claudin-1 and occludin in ileal and colonic mucosa were detected by Western blot. Data were analyzed by 2 × 2 factorial arrangement of traetments using the PROC MIXED. Dietary MFGM had no adverse impact on growth performance or TJ proteins (P > 0.05). Dietary MFGM increased VH (CON: 348.8 and MFGM: 427.0 ± 26.79 µm) and decreased CD (CON: 166.3 and MFGM: 147.4 ± 8.42 µm) in ileum, thereby increasing the VH: CD ratio (CON: 2.2; MFGM: 3.1 ± 0.13; P < 0.05). Further, LPS treatment increased serum I-FABP by 4.4-fold and decreased jejunal mucosal DAO by 0.5-fold compared with saline treatment (P < 0.05). Additionally, there was a 93.1 % increase in colon DAO activity with MFGM diet compared with soy-fed pigs, regardless of LPS challenge (P < 0.05). Dietary MFGM improved neonatal piglet ileum architecture and increased colon DAO activity, potentially attenuating the intestinal barrier disruption following LPS challenge.

The influence of MPL addition on structure, interfacial compositions and physicochemical properties on infant formula fat globules

The lack of milk fat globule membrane phospholipids (MPL) at the interface of infant formula fat globules has an impact on the stability of fat globules, compared to human milk. Therefore, infant formula powders with different MPL contents (0%, 10%, 20%, 40%, 80%, w/w of MPL/whey protein mixture) were prepared, and the effect of interfacial compositions on the stability of globules was investigated. With increasing MPL amount, the particle size distribution had two peaks and returned to a uniform state when 80% MPL was added. At this composition, the MPL at the oil–water interface formed a continuous thin layer. Moreover, the addition of MPL improved the electronegativity and the emulsion stability. In terms of the rheological properties, increasing the concentration of MPL improved the elastic properties of the emulsion and the physical stability of the fat globules, while reducing the aggregation and agglomeration between fat globules. However, the potential for oxidation increased. Based on these results, the interfacial properties and stability on infant formula fat globules was significantly influenced by the level of MPL, which should be considered in the design of infant milk powders.

Heat-stable liposomes from milk fat globule membrane phospholipids for pH-triggered delivery of hydrophilic and lipophilic bioactives

Heat-stable, multivitamin-loaded, and surface-coated liposomes tailored for pH-triggered delivery of both lipophilic and hydrophilic bioactives were synthesized from a cocktail of milk fat globule membrane (MFGM) phospholipids, supercritically extracted from buttermilk powder. Liposomes from MFGM-phospholipids (ML) were fabricated using a novel organic solvent free process based on venturi-based rapid expansion of a supercritical solution. A commercially available pH-responsive polymer, Eudragit® S100 was used to coat ML (Eu-ML). Eu-MLs were heated to 60, 75, and 90 °C for 30 min and were observed to be heat-stable as established by CLSM images, structural characterizations, and encapsulation efficiency measurements. Coating of liposomes with Eudragit® S100 protected encapsulated payload from deleterious gastric environment and facilitated site-specific pH-triggered release in the simulated intestinal condition. This study established a green method to fabricate heat-stable liposomal vehicles for pH-triggered delivery of bioactive compounds with excellent potential for scale-up and applications in the food, pharmaceutical, and cosmetic industries.

Milk fat globule membrane phospholipids modify adhesion of Lactobacillus to mucus-producing Caco-2/Goblet cells by altering the cell envelope

The importance of various Lactobacillus strains and milk components, such as the milk fat globule membrane, has been studied from various perspectives and proven to have a positive role in human health. On one end, lactic acid bacteria produce metabolites with direct effect in the immune system, changes of pH in the gut, and antagonistic substances for pathogenic bacteria as well as competition. On the other end, the milk fat globule membrane improves gastrointestinal status by promoting cell proliferation, epithelial tight junction patterns, and development of intestinal epithelial cells. Interaction between beneficial bacteria and milk fat is a natural occurring phenomenon in dairy products; however, it has not been fully characterized. In this work, we studied the effect of milk phospholipids in the adhesion of Lactobacillus to mucus-producing Caco-2/Goblet cell co-cultures and found that treatment with phospholipids produced bacterial cells with increased surface electronegativity, which was correlated with increased bacterial cells adhered to the intestinal model. Moreover, we utilized an original means of characterizing the adhesion using quartz crystal microbalance. All strains studied, experienced modification of adhesion either physicochemical or kinetic parameters studied. Furthermore, by imaging bacterial cells by electron microscopy, we identified that some strains, such as L. acidophillus and L. casei, metabolized MPL, which improved their adhesion to hydrophilic surfaces such as gold. We identified another group of bacteria, such as L. delbrueckii and L. plantarum, that, instead of metabolizing MPL, kept the phospholipids bound irreversibly to the surface of the cell envelope thus decreasing their adherence to gold surfaces. One of the most important aspects of probiotic lactic acid bacteria —besides survival in the stomach—is the colonization and extended resident time in the intestine to effectively change the gut microbiome. We found that bacterial treatment with milk phospholipids enhances adhesion to intestinal models and will in turn, increase the residence time with the concomitant benefits to the consumer.

Effect of Particle Size and Interface Composition on the Lipid Digestion of Droplets Covered with Membrane Phospholipids

The particle size and fatty acid release of droplets covered with milk fat globule membrane phospholipids with different particle sizes (large/MPL-L; medium/MPL-M; and small/MPL-S) and emulsions with different sources (droplets covered with MPL/MPLs; human milk/HM; and infant formula/IF) were investigated using an infant digestion model. During digestion, droplets exhibited different degrees of aggregation, and the order of the particle size was MPL-L > MPL-M > MPL-S. MPL-M and MPL-S were significantly higher than MPL-L in the release of free fatty acids. No significant difference was observed in the FFA release rate between MPLs and HM. However, the rate was significantly higher than that of IF in the intestinal stage. Compared to IF, a higher content of long-chain polyunsaturated fatty acids and a lower content of saturated fatty acid were observed in MPLs and HM.

Arachidonic Acid in Human Milk.

Breastfeeding is universally recommended as the optimal choice of infant feeding and consequently human milk has been extensively investigated to unravel its unique nutrient profile. The human milk lipid composition is unique and supplies specifically long-chain polyunsaturated fatty acids (LC-PUFAs), in particular, arachidonic acid (ARA, 20:4n–6) and docosahexaenoic acid (DHA, 22:6n–3). Arachidonic acid (ARA) is the most predominant long-chain polyunsaturated fatty acid in human milk, albeit at low concentrations as compared to other fatty acids. It occurs predominantly in the triglyceride form and to a lesser extent as milk fat globule membrane phospholipids. Human milk ARA levels are modulated by dietary intake as demonstrated by animal and human studies and consequently vary dependent on dietary habits among mothers and regions across the globe. ARA serves as a precursor to eicosanoids and endocannabinoids that also occur in human milk. A review of scientific and clinical studies reveals that ARA plays an important role in physiological development and its related functions during early life nutrition. Therefore, ARA is an important nutrient during infancy and childhood and, as such, appropriate attention is required regarding its nutritional status and presence in the infant diet. Data are emerging indicating considerable genetic variation in encoding for desaturases and other essential fatty acid metabolic enzymes that may influence the ARA level as well as other LC-PUFAs. Human milk from well-nourished mothers has adequate levels of both ARA and DHA to support nutritional and developmental needs of infants. In case breastfeeding is not possible and infant formula is being fed, experts recommend that both ARA and DHA are added at levels present in human milk.

Synthesis of multivitamin-loaded heat stable liposomes from milk fat globule membrane phospholipids by using a supercritical-CO2based system

Inspired by the heat stability of milk, where fat globules are coated by the milk fat globule membrane (MFGM), heat stable liposomes loaded with multivitamins were successfully synthesized from MFGM phospholipid concentrate.

Impact of technological processes on buffalo and bovine milk fat crystallization behavior and milk fat globule membrane phospholipids profile

The effects of milk technological processes (pasteurization, homogenization, and freeze-drying) on the crystallization and melting behaviors of buffalo and bovine milk fat were investigated. As well, the profile of milk phospholipids was identified. Differential scanning calorimetry was used to evaluate the variations in the thermal behaviors of milk fat. Milk phospholipids were purified by the solid-phase extraction method, and then identified by using an ultra-performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry. Seven classes of phospholipids (including phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, sphingomyelin, and lysophosphatidylcholine) were identified. Milk processing affected both the thermal characteristics and the profile of phospholipids, since the content of phosphatidylcholine increased after the homogenization of pasteurized milk. The finding of this study could provide a valuable knowledge to dairy industry and the related applications since it highlights the variations in milk fat crystallization and melting behaviors, and the profile of phospholipids throughout a series of technological treatments.

Curcumin liposomes prepared with milk fat globule membrane phospholipids and soybean lecithin

Using thin film ultrasonic dispersion method, the curcumin liposomes were prepared with milk fat globule membrane (MFGM) phospholipids and soybean lecithins, respectively, to compare the characteristics and stability of the 2 curcumin liposomes. The processing parameters of curcumin liposomes were investigated to evaluate their effects on the encapsulation efficiency. Curcumin liposomes were characterized in terms of size distribution, ζ-potential, and in vitro release behavior, and then their storage stability under various conditions was evaluated. The curcumin liposomes prepared with MFGM phospholipids had an encapsulation efficiency of about 74%, an average particle size of 212.3nm, and a ζ-potential of −48.60mV. The MFGM liposomes showed higher encapsulation efficiency, smaller particle size, higher absolute value of ζ-potential, and slower in vitro release than soybean liposomes. The retention rate of liposomal curcumin was significantly higher than that of free curcumin. The stability of the 2 liposomes under different pH was almost the same, but MFGM liposomes displayed a slightly higher stability than soybean liposomes under the conditions of Fe3+, light, temperature, oxygen, and relative humidity. In conclusion, MFGM phospholipids have potential advantages in the manufacture of curcumin liposomes used in food systems.

Encapsulation of bioactive salmon protein hydrolysates with chitosan-coated liposomes

A nano-encapsulation strategy for a chitosan-coated liposomal oral delivery system using milk fat globule membrane (MFGM) phospholipids for the entrapment of antidiabetic peptides prepared from Atlantic salmon (Salmo salar) protein hydrolysates (SPH) was developed. The size, zeta potential, entrapment efficiency, stability during freeze-drying freeze thawing, and long-term storage abilities of the nano-capsules were investigated as a function of concentrations of phospholipid and chitosan coating material. Chitosan coating greatly improved the stability of liposomes. The maximum encapsulation efficiency (71.3%) and physical stability were achieved with 10% MFGM phospholipid and 0.4% chitosan. Chitosan coating significantly prolonged the release of SPH in simulated biological fluids. In conclusion, liposomes with “optimal” chitosan coating-concentration show great promise as a potential new delivery system for protein hydrolysates.

Storage Stability and Physical Characteristics of Tea-Polyphenol-Bearing Nanoliposomes Prepared with Milk Fat Globule Membrane Phospholipids

The objective of this work was to better understand the functional properties of milk phospholipids when used as ingredients to prepare liposomes. Liposomal dispersions (10%) were prepared using high-pressure homogenization, and their physical properties as well as their ability to encapsulate tea polyphenols were investigated. The extent of encapsulation, measured by HPLC, increased with tea polyphenol concentration up to about 4 mg·mL(-1). At polyphenol concentrations ≥ 6 mg·mL(-1), the liposome dispersions were no longer stable. The influence of pH (3-7), storage temperature (room temperature or refrigeration), and addition of sugars (0-15%) were studied for liposomes containing 4 mg·mL(-1) polyphenols. The liposomal dispersions were also stable in the presence of peptides. The storage stability of the systems prepared with milk phospholipids was compared to that of liposomes made with soy phospholipids. Soy liposomes were smaller in size than milk phospholipid liposomes, the encapsulation efficiency was higher, and the extent of release of tea polyphenols during storage was lower for milk phospholipid liposomes compared to soy liposomes. The results suggest that milk phospholipids could be employed to prepare tea-polyphenol-bearing liposomes and that the tea catechins may be incorporated in the milk phospholipid bilayer more efficiently than in the case of a soy phospholipid bilayer.

Structure and integrity of liposomes prepared from milk- or soybean-derived phospholipids during in vitro digestion

Crude liposomes and nanoliposomes, respectively, formed both from a milk fat globule membrane (MFGM) phospholipid fraction and from soybean phospholipid were prepared by thin layer dispersion and dynamic high pressure microfluidization methods. The structure and the integrity of the liposomes were evaluated in terms of average diameter, zeta potential, microstructure, lipolysis, and membrane permeability during in vitro digestion as a function of time. The physical and chemical properties of the liposomes were little influenced in SGF containing pepsin. However, the liposomes exhibited lower stability in simulated intestinal fluid (SIF) than in simulated gastric fluid (SGF). The liposomes obtained from the MFGM phospholipid fraction were more stable than the soybean-based liposomes, with less change in average diameter, surface charge, morphology, and free fatty acid release, and better membrane integrity during digestion in SIF. However, no differences in the stability between nanoliposomes and crude liposomes were observed during digestion. These results systematically demonstrated the relationship between the stability of liposomes and in vitro digestion, and provided some information for further developing more stable liposomes in the gastrointestinal tract.

Composition, Structure, and Bioactive Components in Milk Fat Globule Membrane

A unique biophysical membrane which surrounds the milk fat globules is called the milk fat globule membrane (MFGM). Various researches were studied about origin, composition, structure and bioactive components of MFGM. Bioactive protein components of MFGM play an important beneficiary function such as defense mechanism in new born. Among the bioactive lipid components from MFGM phospholipids showed health enhancing functions. The phospholipids also help in the production of certain dairy product from deterioration. MFGM phospholipids also showed antioxidant activity in some dairy products such as butter and ghee produced from milk of buffalo. Based on the beneficial effects, researchers developed MFGM as functional ingredients in various food products. This current review focuses on health enhancing function of MFGM and its components in various dairy products.

Use of ultrafiltration and supercritical fluid extraction to obtain a whey buttermilk powder enriched in milk fat globule membrane phospholipids

Whey buttermilk, a by-product from whey cream processing to butter, is rich in milk fat globule membrane (MFGM) constituents, which have technological and potential health properties. The objective of this work was to produce a dairy ingredient enriched in MFGM material, especially phospholipids, from whey buttermilk. Whey buttermilk was concentrated by ultrafiltration (10×) and subsequently diafiltered (5×) (10 kDa molecular mass cutoff membrane) at 25 °C and the final retentate was spray-dried. The whey buttermilk powder was submitted to supercritical extraction (350 bar, 50 °C) using carbon dioxide. The membrane filtration removed most of the lactose and ash from the whey buttermilk, and the supercritical extraction extracted exclusively non-polar lipids. The final powder contained 73% protein and 21% lipids, of which 61% were phospholipids. This ingredient, a phospholipids-rich dairy powder, could be used as an emulsifier in different food systems.

The properties of liposomes produced from milk fat globule membrane material using different techniques

The isolation of milk fat globule membrane (MFGM) material from buttermilk on a commercial scale has provided a new ingredient rich in phospholipids and sphingolipids. In the pharmaceutical and cosmetic industries, highly purified phospholipids extracted from soya oil or egg yolk are used to produce liposomes. Liposomes are spherical structures consisting of one or more phospholipid bilayers enclosing an aqueous core. They may be used for the entrapment and controlled release of drugs or nutraceuticals, as model membranes or cells, and even for specialist techniques such as gene delivery. There are many potential applications for liposomes in the food industry, ranging from the protection of sensitive ingredients to increasing the efficacy of food addi- tives. Our previous work compared the structure and properties of liposomes prepared from a milk fat globule membrane (MFGM) fraction and soya phospholipid material using a high-pressure ho- mogenizer (Microfluidizer). These results identified some potential advantages in the use of MFGM phospholipids for the manufacture of liposomes for use in food systems. This paper compared the general structure and properties of liposomes prepared from the same MFGM phospholipid mate- rial using three different techniques - microfluidization, the traditional thin-film hydration and the heating method. The thin-film hydration technique required the use of organic solvents, while the other two methods do not involve any non food-safe chemicals. The liposomes prepared by both microfluidization and the heating method had high entrapment efficiencies. Liposomes produced via microfluidization tended to be significantly smaller than those produced by the other methods, with a narrower size distribution, and a higher proportion of unilamellar vesicles. There did not seem to be any advantages in the use of the thin-film hydration method, opening the door to the use of food-safe methods for liposome production. liposome / milk fat globule membrane / phospholipid / microfluidization

Physical Stability of Liposomes Prepared from Milk Fat Globule Membrane and Soya Phospholipids

Previous research has shown that liposomes prepared from a milk fat globule membrane (MFGM) phospholipid fraction had a significantly higher phase transition temperature, thicker membrane, and lower membrane permeability than liposomes prepared from soya phospholipid material. Subsequent investigations into the relative stability of the two liposome dispersions have found that the MFGM phospholipid liposomes are more stable than their soya counterparts in a range of pH conditions, at a variety of storage and processing temperatures, and in the presence of mono- and divalent cations. These results illustrate some potential advantages in the use of MFGM phospholipids for the manufacture of liposomes for use in food systems.

Preparation of Liposomes from Milk Fat Globule Membrane Phospholipids Using a Microfluidizer

The isolation of milk fat globule membrane (MFGM) material from buttermilk on a commercial scale has provided a new ingredient rich in phospholipids and sphingolipids. An MFGM-derived phospholipid fraction was used to produce liposomes via a high-pressure homogenizer (Microfluidizer). This technique does not require the use of solvents or detergents, and is suitable for use in the food industry. The liposome dispersion had an average hydrodynamic diameter of 95nm, with a broad particle-size distribution. Increasing the number of passes through the Microfluidizer, increasing the pressure, or reducing the phospholipid concentration all resulted in a smaller average liposome diameter. Changing these variables did not have a significant effect on the polydispersity of the dispersion. Electron microscopy showed that the dispersions formed had a range of structures, including unilamellar, multilamellar, and multivesicular liposomes. The composition of the MFGM phospholipid material is different from that of the phospholipids usually used for liposome production in the pharmaceutical and cosmetic industries. The MFGM-derived fraction comprises approximately 25% sphingomyelin, and the fatty acids are primarily saturated and monounsaturated. These differences are likely to affect the properties of the liposomes produced from the phospholipid material, and it may be possible to exploit the unique composition of the MFGM phospholipid fraction in the delivery of bioactive ingredients in functional foods.

Differently sized native milk fat globules separated by microfiltration: fatty acid composition of the milk fat globule membrane and triglyceride core

The aim of this study was to characterize the fatty acid composition of the core and membrane of differently sized milk fat globules separated by microfiltration, which can now be used to manufacture dairy products. Native milk fat globules of various mean diameters, ranging from d 43 = 2.3 μm to 8.0 μm, were obtained using microfiltration of raw whole milk. After milk fat globule washing, the milk fat globule membrane (MFGM) and the triglyceride core (TC) were separated by manual churning. After total lipid extraction from each fraction, their fatty acid composition was characterized using methyl ester analysis by gas chromatography. Regardless of season, no significant differences were observed in the fatty acid composition of the MFGM phospholipids. Conversely, significant differences were found in the fatty acid composition of TC; particularly, small fat globule TC contained more medium-chain fatty acids and less stearic acid than large fat globule TC. These results show that the previously observed differences in total fatty acid composition among differently sized milk fat globules are due to their triglyceride composition and MFGM amount rather than to the composition of the MFGM.