Addition Of Lactoferrin Research Articles

Encapsulation of gallic acid in alginate/lactoferrin composite hydrogels: Physical properties and gallic acid diffusion

Alginate hydrogel is commonly utilized to encapsulate hydrophilic polyphenols because of its biocompatibility and flexibility. However, encapsulation with neat alginate gel is prone to uncontrolled compound diffusion during gel formation and storage. Previous research have demonstrated the electrostatic complexation of lactoferrin and alginate, nonetheless, no study has been reported on the combination of these polymers as hydrophilic polyphenol encapsulation agent. Therefore, this research aims to explore the encapsulation of gallic acid (GA), a representative of water-soluble polyphenol, in the lactoferrin-alginate composite hydrogel. The composite hydrogel of sodium alginate-lactoferrin (S-L) was prepared in solid-basis ratios of 1:0, 1:1, 2:0, 2:1, 2:2, and 2:3. The evaluation was conducted on the alginate-lactoferrin mixture viscosity, gel syneresis, gel strength and stiffness, dried gel rehydration properties, porosity, GA diffusion, and gel microstructure. Images of SEM and CLSM microscopy revealed relatively compact structures created by evenly distributed lactoferrin in the hydrogel system. According to the gel electrophoresis analysis, composite alginate-lactoferrin gel hindered the diffusion of compounds with molecular weight >50 kDa. The addition of lactoferrin to the GA-loaded alginate composite hydrogel significantly increased the retention of GA (87.1–92.2%) in the gel. The GA-loaded composite hydrogel with the alginate to lactoferrin ratio of 2:1 and 2:2 (pH 5.24–5.43) exhibited significantly better (P > 0.05) gallic acid retention compared to the neat alginate hydrogel (pH 5.16) in their natural pH. FTIR analysis revealed a hydrogen bond between lactoferrin and the gallic acid phenolic group, while the zeta-potential analysis showed the intermolecular interaction between positive-charged lactoferrin and negative-charged alginate. This work provides important information for the development of hydrogel-based polyphenol encapsulating materials.

Properties of Yoghurt Fortified in Lactoferrin with Effect of Storage Time.

The stability of fortified yoghurts during refrigerated storage is important for industry and the consumer. The aim of the study was to evaluate the nutritional value, microbiological quality, organoleptic properties, and structure of natural yoghurts made with the addition of lactoferrin during refrigerated storage. In this study, we produced natural yoghurts fortified in lactoferrin, using YC-X11 yoghurt starter culture based on Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Physicochemical (acidity, nutritional value and structure) as well as microbiological and organoleptic changes occurring during 28-days refrigerated storage were determined. Storage research made it possible to determine the direction of changes taking place in the products. The analysed parameters did not differ statistically significantly between the control yoghurts and those with the addition of lactoferrin. Textural and rheological studies also shown that the addition of lactoferrin did not significantly change the structure of the yoghurt. The yoghurts were characterized by high sanitary and hygienic quality during the whole refrigerated storage. Lactoferrin has a positive effect on the durability of the product.

Synergistic effect of lactoferrin and red yeast rice on the quality characteristics of novel plant-based meat analog patties

This study reports physicochemical, textural characteristics, sensory properties, and fatty acid profile of plant-based meat analog (PBMA) patties formulated with lactoferrin (LTF), red yeast rice (RYR), and the combination of LTF and RYR compared to control (beef). The comparison revealed that the raw patties in the treated groups were lighter (42–46) in color (p < 0.05) and displayed moderate yellowness (17–20) than control. Conversely, in cooked patties, all color indices decreased meaningfully (p < 0.05) compared to control. The proximate composition validated higher moisture (60–61%), protein (22–25%), and fat (6–7%) in comparison to control. Whereas, shrinkage percentage (2.35–5.33%) and cooking yield (CY%) (2.24–2.11%) reduced considerably (p < 0.05) in treated samples than in control. The electronic tongue system (ETS) taste traits of sourness and saltiness were negatively affected by the addition of LTF and RYR, while the umami taste attribute displayed a positive tendency in the treated samples compared to the control. Collectively, the incorporation of natural colorants substantially affected the two major clusters of fatty acid profiles. The principal component analysis model showed clear separation (Dim1 and Dim2) in taste and textural attributes, which necessitates a further investigation to develop the superior quality of plant-based meat products.

Activity of Phage-Lactoferrin Mixture against Multi Drug Resistant Staphylococcus aureus Biofilms.

Biofilms are complex bacterial structures composed of bacterial cells embedded in extracellular polymeric substances (EPS) consisting of polysaccharides, proteins and lipids. As a result, biofilms are difficult to eradicate using both mechanical methods, i.e., scraping, and chemical methods such as disinfectants or antibiotics. Bacteriophages are shown to be able to act as anti-biofilm agents, with the ability to penetrate through the matrix and reach the bacterial cells. However, they also seem to have their limitations. After several hours of treatment with phages, the biofilm tends to grow back and phage-resistant bacteria emerge. Therefore, it is now recommended to use a mixture of phages and other antibacterial agents in order to increase treatment efficiency. In our work we have paired staphylococcal phages with lactoferrin, a protein with proven anti-biofilm proprieties. By analyzing the biofilm biomass and metabolic activity, we have observed that the addition of lactoferrin to phage lysate accelerated the anti-biofilm effect of phages and also prevented biofilm re-growth. Therefore, this combination might have a potential use in biofilm eradication procedures in medical settings.

Lactoferrin improves the immune response and resistance of silver carp, a hematological, light (histochemical and immunohistochemical), fluorescent, and scanning electron microscopic study.

Lactoferrin is a multifunctional protein that has roles in iron metabolism, cell generation and differentiation, and antibacterial, antiviral, and antiparasitic activity. The study aim was to evaluate the effect of dietary lactoferrin powder on the innate immune response and disease resistance of silver carp (Hypophthalmichthys molitrix) infected with Vibrio vulnificus. The study also investigates immune cells in different organs including gills, intestine, spleen, liver, and pancreas using light, scan electron microcopy. Two diet regimes, a basic diet and a 600-mg/kg bovine lactoferrin-supplemented diet were used. The fish were fed for 30 successive days. We included 270 apparently healthy silver carp (H. Molitrix) with an average body weight of 10± 2g/fish. Addition of lactoferrin to the diet led to an increase in the activity of non-specific immune parameters; lymphocytes, and in the monocytes percentage, and in total protein, serum globulin, and albumin, with a corresponding increase in phagocytic activities and indices. The histological analysis revealed increase immune cells such as lymphocytes in intestine, rodlet cells in liver, gills, and pancreas, and the macrophages in liver, pancreas as well as spleen. The lactoferrin-supplemented diet significantly increased survival and disease resistance following V. vulnificus challenge, which seemed to correspond to an enhancement of non-specific immune functions in H. molitrix. RESEARCH HIGHLIGHTS: The study evaluates the effect of dietary lactoferrin powder on the innate immune response and disease resistance of silver carp (Hypophthalmichthys molitrix) infected with Vibrio vulnificus. The study also investigates immune cells in different organs including gills, intestine, spleen, liver, and pancreas using light, scan electron microcopy. Lactoferrin increases the activity of nonspecific immune parameters; lymphocytes, and in the monocytes percentage, and in total protein, serum globulin, and albumin, with a corresponding increase in phagocytic activities and indices. The histological analysis revealed increase immune cells such as lymphocytes in intestine, rodlet cells in liver, gills, and pancreas, and the macrophages in liver, pancreas as well as spleen.

Addition of lactoferrin and substance P in a chitin/PLGA-CaSO4 hydrogel for regeneration of calvarial bone defects

Calcium-based injectable hydrogels with various bioactive active molecules possess a great potential for bone regeneration. Herein, we have synthesized a chitin-PLGA-calcium sulfate hydrogel (CSG) containing bioactive molecules - lactoferrin (LF) and substance P (SP). SEM and XRD analysis revealed that CS crystal growth was altered with the addition of LF. Rheological measurements indicated that the injectability of the hydrogels was maintained after the addition of LF, however, there was a reduction in storage modulus after LF addition. The addition of LF increased stem cell proliferation whereas, SP enhanced the cell migration. Osteogenic gene expression revealed that LF concentration at 25 μg/mg of CSG was optimal for a favourable outcome. To this optimized LF containing CSG, SP was incorporated and 0.05 μg/mg was found to be most effective (CSG-L3S2) in vitro studies. Further, the μ-CT and histological studies confirmed that CSG-L3S2 showed enhanced bone regeneration compared to the controls in critical-sized calvarial defect of mice. Thus the results indicate that a combination of the chemotactic agent (SP), pleiotropic growth protein (LF), and CS in the chitin-PLGA hydrogel could be a promising approach for non-load bearing bone defects.

Effects of exogenous lactoferrin on characteristics and functions of bovine epididymal, ejaculated and frozen-thawed sperm.

The purpose of this study was to investigate effects of addition of lactoferrin on characteristics and functions of bovine epididymal, ejaculated, and frozen-thawed sperm. The addition of lactoferrin was significantly (p<.05) effective on increasing values of progressive motility, straightness, and linearity in caput epididymal sperm and values of motility in cauda epididymal sperm. When ejaculated sperm were incubated in capacitation medium, percentages of motile and progressively motile sperm decreased largely within the first period of 30min, followed by only minor changes. However, the addition of lactoferrin significantly lessened the early decreases of these parameters and additionally promoted capacitation-dependent changes of chlortetracycline staining patterns (from F pattern to B pattern). In other experiments, when ejaculated sperm were exposed to oxidative stress with 100-µM H2 O2 , the addition of lactoferrin partially protected them from dysfunction of flagellar movement and loss of progressive movement. In final experiments with frozen-thawed samples incubated in the capacitation medium, the addition of lactoferrin effectively survived dying sperm and suppressed occurrence of sperm agglutination. These results may suggest biological and biotechnological potentials of lactoferrin for modulation of bovine sperm viability, motility, capacitation state, and preservation in vitro.

The Combination of Two Bioactive Constituents, Lactoferrin and Linolenic Acid, Inhibits Mouse Xenograft Esophageal Tumor Growth by Downregulating Lithocholyltaurine and Inhibiting the JAK2/STAT3-Related Pathway.

The addition of lactoferrin and three unsaturated fatty acids, oleic acid, docosahexaenoic acid (DHA), and linolenic acid, to dairy products was approved in recent years. Research into the biological activities of lactoferrin and these three unsaturated fatty acids has revealed anti-inflammatory, antiviral, antioxidant, antitumor, antiparasitic, and antibiotic effects. However, investigations and comparisons of lactoferrin + oleic acid/DHA/linolenic acid combinations in an esophageal cancer cell model and in xenograft tumor models have not been extensively reported, and the related mechanism of these combinations remains elusive. In the present study, the effects of lactoferrin and the three fatty acids on KYSE450 cell viability, migration, and invasion were investigated to choose the proper doses and effective combination in vitro. A tumor-bearing nude mouse model was established to investigate the role of selected combinations in inhibiting esophageal tumor formation in vivo. Metabonomics detection and data analysis were performed to screen special metabolites and related pathways, which were validated by western blotting. The results demonstrated that lactoferrin, the three unsaturated fatty acids, and their combinations inhibited the viability, migration, and invasion of KYSE450 cells and induced apoptosis and the lactoferrin + linolenic acid combination exhibited the strongest activity in suppressing KYSE450 tumor formation in vivo. The lactoferrin + linolenic acid combination inhibited phosphorylation in the JAK2/STAT3-related pathway by downregulating the special metabolite lithocholyltaurine, thereby suppressing formation of KYSE450 tumors.

Influence of interfacial compositions on the microstructure, physiochemical stability, lipid digestion and β-carotene bioaccessibility of Pickering emulsions

Novel Pickering emulsions were co-stabilized by solid particles and two types of emulsifiers (lactoferrin or rhamnolipid) for delivery of β-carotene. The influence of the particle-surfactant or particle-protein mixed interface on novel Pickering emulsions was investigated. The droplet size of the particles and lactoferrin co-stabilized Pickering emulsion was increased from 13.34 ± 0.03 μm to 20.70 ± 0.09 μm with the rise in lactoferrin level from 0.10% to 1.25% (w/v), but the droplet size of the particles and rhamnolipid co-stabilized Pickering emulsion was increased initially from 17.61 ± 0.10 μm to 38.24 ± 0.11 μm with the increase in rhamnolipid level from 0.10% to 0.75% (w/v) and then decreased to 15.17 ± 0.25 μm at 1.25% of rhamnolipid. Compared to the emulsion stabilized by particles solely, the stability of Pickering emulsions co-stabilized by particles and emulsifiers, and the β-carotene entrapment were improved under environmental stresses. Presence of lactoferrin or rhamnolipid might induce the competitive displacement, adsorption onto particle surface or multilayer deposition at the interface, which were observed through confocal laser scanning microscope and cryo-scanning electric microscopy. Furthermore, the addition of lactoferrin or rhamnolipid inhibited the release of free fatty acids from in vitro digestion effectively, thereby reducing the bioaccessibility of β-carotene slightly. These findings facilitated to understand the effect of interfacial compositions on the stability and digestion fate of the nutraceutical emulsions co-stabilized by particles and protein or surfactant.

Biopreservative Effect of Lactoferrin Against Foodborne Pathogens Inoculated in Egyptian Soft Cheese “Karish Cheese”

Lactoferrin is a natural component that is used as a food additive to inhibit the growth of pathogenic microorganisms. This study was designed to evaluate the antibacterial effect of lactoferrin against E. coli O157:H7, S. aureus L. monocytogenes, and B. cereus inoculated in one of the most popular Egyptian soft cheese “Karish cheese”. The sensory properties including appearance, odor and flavor of the cheese, to which lactoferrin was added, were evaluated. The results of sensory evaluation revealed that the samples treated with the different concentrations of lactoferrin were generally acceptable and did not negatively affected its sensorial properties. Moreover, the samples treated with high concentration of lactoferrin were given high score. Antibacterial effect of 1%, 2% and 4% concentrations of bovine lactoferrin against the foodborne pathogens were tested in vitro using agar well diffusion test to determine the appropriate concentration to be applied in cheese. The results showed that lactoferrin 4% concentration was the most effective concentration against all the examined foodborne pathogens, E. coli O157:H7, S. aureus L. monocytogenes, and B. cereus. The antibacterial effect of 4% lactoferrin against the before mentioned foodborne pathogens in Karish cheese was investigated and it could inhibit them during the cold storage period. This study highlighted that lactoferrin improved sensory properties of Karish cheese experimentally inoculated with some food borne pathogens such as E. coli O157:H7, S. aureus L. monocytogenes, and B. cereus, in addition to extending its shelf life at cold storage. Therefore, the addition of lactoferrin to Karish could improve its microbiological safety.

Effect of Different Culture Conditions on In vitro Development and Quality of Buffalo Embryos

This work was carried out to evaluate the quality and the developmental competence of the in vitro buffalo embryos produced under different culture conditions. Oocytes were matured in TCM 199 for 24 h at 38.5°C and 5% CO2. After maturation, oocytes were in vitro fertilized by frozen thawed sperms capacitated in vitro by caffeine at final concentration 2 million sperm /ml. The sperm-oocytes were incubated in TALP medium for 24 hrs. Fertilized oocytes were cultured in Cell free media (CR1aa) or with cumulus or oviductal cell monolayers (Experiment 1), CR1aa with addition of (10, 20, 50 and 100 µg/mL) lactoferrin (Experiment 2) inside CO2 incubator with 5% CO2 at 38°C. The cleavage, morula, blastocyst rates were recorded and embryo cell number and mitochondrial function were evaluated. Addition of bovine oviductal cell monolayer (BOCM) to culture media significantly increased the morula and the blastocyst compared with cell free media and media with cumulus cell monolayer and also significantly increased the cell number and the mitochondrial function of in vitro produced buffalo embryos. Addition of lactoferrin by concentration 50µg/ml to culture media resulted in a significant increase in rates of cleavage; morula and blastocyst compared with control or other concentrations and also resulted in an increase in the cell number and the mitochondrial function of in vitro produced buffalo embryos. It could be concluded that, addition of oviductal cells (monolayer) or lactoferrin to culture medium significantly improved the quality of in vitro produced buffalo embryos.

Lactoferrin as an added therapy in the treatment of Helicobacter pylori

Background Bovine lactoferrin (bLF), an iron-binding glycoprotein, is a nonenzymatic antioxidant found in the whey fraction of fermented milk as well as in colostrum. The possibility that bLF may help improve the Helicobacter pylori eradication rate was first conceived in 1997 when an in-vitro study by Yamazaki and colleagues showed that bLF was bactericidal to H. pylori in Brucella broth. Later in-vitro studies have confirmed the same and yielded evidence of the possible mechanism of the bactericidal action of bLF, relating it to the high iron-binding affinity and prevention of iron utilization by H. pylori. Lactoferrin is used for treating stomach and intestinal ulcers, diarrhea, and hepatitis C. Aim The aim of the study was to verify the value of adding lactoferrin to the treatment of H. pylori infection. Patients and methods This study was conducted on 50 patients selected from Internal Medicine Department, Benha University Hospital. They were divided into group 1, which included 25 patients who were treated with traditional therapy (clarithromycin, omeprazole, amoxicillin, or metronidazole), and group 2, who were treated by traditional therapy plus lactoferrin (pravotin) for 1 week. H. pylori stool antigen testing was performed for both groups before and after therapy to assess the response to therapy. Results were statistically analyzed. Results Our results show that addition of lactoferrin improves the H. pylori infection eradication rate.

Formation of interfacial milk protein complexation to stabilize oil-in-water emulsions against calcium

Oil-in-water emulsions (30% soya oil) stabilized by sodium caseinate or whey protein were destabilized in the presence of calcium, whereas lactoferrin-stabilized emulsion was stable under same conditions. At pH7, addition of lactoferrin to the caseinate or whey protein-stabilized emulsions led to association of lactoferrin with adsorbed proteins by electrostatic interactions, hence resulted in the formation of lactoferrin-caseins/whey protein interfacial complexes at the interface of the emulsion droplets. The stability of the interfacial complexes-coated emulsion droplets in the presence of Ca2+ was examined by determination of particle size, zeta-potential and microstructure as a function of Ca2+ concentration and added lactoferrin concentration. For emulsions made with 1% w/w caseinate or WPI, the stability of emulsions in the presence of 20mM CaCl2 has been improved markedly after addition of 0.2% w/w lactoferrin, suggesting the small amount of lactoferrin on the interface can largely enhance the calcium stability of milk proteins-coated emulsion droplets. Steric repulsion between the interfaces of droplets produced by the large lactoferrin molecules on the interface was considered to contribute the preventing the flocculation caused by Ca2+ binding and reduction in the electrostatic repulsion between the emulsion droplets.

Interaction of Lactoferrin and Lysozyme with Casein Micelles

On addition of lactoferrin (LF) to skim milk, the turbidity decreases. The basic protein binds to the caseins in the casein micelles, which is then followed by a (partial) disintegration of the casein micelles. The amount of LF initially binding to casein micelles follows a Langmuir adsorption isotherm. The kinetics of the binding of LF could be described by first-order kinetics and similarly the disintegration kinetics. The disintegration was, however, about 10 times slower than the initial adsorption, which allowed investigating both phenomena. Kinetic data were also obtained from turbidity measurements, and all data could be described with one equation. The disintegration of the casein micelles was further characterized by an activation energy of 52 kJ/mol. The initial increase in hydrodynamic size of the casein micelles could be accounted for by assuming that it would go as the cube root of the mass using the adsorption and disintegration kinetics as determined from gel electrophoresis. The results show that LF binds to casein micelles and that subsequently the casein micelles partly disintegrate. All micelles behave in a similar manner as average particle size decreases. Lysozyme also bound to the casein micelles, and this binding followed a Langmuir adsorption isotherm. However, lysozyme did not cause the disintegration of the casein micelles.

Effects of Lactoferrin and Lysozyme on Ciliary Motility in Rabbit Tracheal Epithelium

We studied the effects of lactoferrin and lysozyme on ciliary beat frequency (CBF) of rabbit cultured tracheal epithelium by a photoelectric method in-vitro. Lactoferrin (50 mg mL−1) and lysozyme (100 mg mL−1) elicited rapid increases in CBF from 13·3 ± 0·3 to 15·9 ± 0·4 Hz (mean ± s.e., P<0·01) and 13·0 ± 0·4 to 17·2 ± 0·5 Hz (P<0·01), respectively, followed by sustained increases which were maintained at least for 30 min. These effects were dose dependent. The maximal increases from the baseline values (ΔCBFmax) and the concentrations required to produce a half-maximal responses (EC50) were 32·4 ± 5·3% and 14·9 ± 2·8 mg mL−1, respectively, for lactoferrin, and 38·0 ± 4·4% and 2·3 ± 0·3 mg mL−1 for lysozyme. The increases in CBF produced by lactoferrin and lysozyme were not affected by pretreatment of cells with propranolol or Ca2+-free medium. Intracellular cAMP levels remained unchanged after the addition of lactoferrin and lysozyme. These results suggest that lactoferrin and lysozyme stimulate airway epithelial ciliary motility, thereby playing a role in the regulation of mucociliary transport in the respiratory tract.

Effects of lactoferrin on the differentiation of pluripotent mesenchymal cells

Lactoferrin accelerates bone formation, but the precise cellular mechanism behind this is still unclear. We examined the effect of lactoferrin on the differentiation of pluripotent mesenchymal cells using a typical pluripotent mesenchymal cell line, C2C12. Cells were cultured in low-mitogen differentiation medium to induce cell differentiation, with or without the addition of lactoferrin. The cell lineage was determined by alkaline phosphatase (ALPase) activity, mRNA expression of cellular phenotype-specific markers using real-time polymerase chain reaction (PCR), and protein synthesis using Western blotting. The expression of low-density lipoprotein lipase receptor-related proteins (LRPs) 1 and 2, both lactoferrin receptors, was determined by reverse transcription-PCR. ALPase activity increased after the addition of lactoferrin. The mRNA expression of Runx2, osteocalcin, and Sox9 increased markedly as a result of lactoferrin treatment, whereas the expression of MyoD, desmin, and PPARgamma decreased significantly. Western blots showed that lactoferrin stimulation increased Runx2 and Sox9 proteins, whereas it decreased MyoD and PPARgamma synthesis. C2C12 cells expressed the LRP1 lactoferrin receptor. These results indicate that lactoferrin treatment converts the differentiation pathway of C2C12 cells into the osteoblastic and chondroblastic lineage.

Lactoferrin Inhibits the Acquisition of Dry-Resistance of &lt;i&gt;Salmonella&lt;/i&gt; spp.

An assay method was established for estimation of dry-resistance of Salmonella. Environmental isolates of Salmonella enterica spp., including S. Enteritidis were grown to the logarithmic phase, washed and re-suspended in saline or Luria-Bertani (LB) medium, followed by drying overnight in an automatic dry-keeper at room temperature. The dried bacteria were recovered by mixing with ice-cold PBS, suspended, and examined for viability by colony-forming activity. A pathogenic clone of S. Enteritidis, SECI#15-1, was not viable in saline alone but maintained its viability in LB medium, suggesting it requires nutrients for the acquisition of dry-resistance. Addition of lactoferrin or apolactoferrin to the bacterial suspension in 20% LB medium prior to the dry-protocol decreased the viability of SECI#15-1 in a dose-dependent manner. However, lactoferrin showed no effect on the growth of SECI#15-1 in liquid culture with LB or M9 medium, suggesting that it exerts bactericidal effects under dry but not under wet conditions. Besides, Salmonella spp. other than S. Emteritidis, such as S. Typhimurium, S. Oranienburg, S. Weltevreden, S. Johannesburg, and S. lnfantis, also showed dry-resistance, which was significantly inhibited by lactoferrin and almost entirely by apolactoferrin. These results suggest that lactoferrin inhibits the acquisition of dry-resistance by Salmonella spp., suggesting that there is a possible use for lactoferrin in the control of Salmonella food-poisoning as an additive in dry food.

In vitro effects of lactoferrin on lipopolysaccharide-induced proliferation, gene expression, and prostanoid production by bovine peripheral blood mononuclear cells

To evaluate the effect of lactoferrin on lipopolysaccharide (LPS)-induced proliferation of bovine peripheral blood mononuclear cells (PBMCs), gene expression of inflammatory mediators, and production of prostanoids in vitro. PBMCs isolated from 15 Holstein bull calves. Mixed populations of PBMCs were isolated by differential centrifugation. Proliferation assays were conducted in 96-well plates designed to allow addition of lactoferrin (200 ng/mL) with and without LPS (1 microg/mL) in a checkerboard design. Incorporation of 3H-thymidine was used to determine proliferation of PBMCs. Prostaglandin E2 production was determined in culture-conditioned medium by use of enzyme immunoassay. Effects of lactoferrin on LPS-induced gene expression of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 were monitored by use of PCR assays. Lactoferrin supplementation significantly reduced LPS-induced incorporation of 3H-thymidine and production of prostaglandin E2 by PBMCs. Lactoferrin reduced LPS-induced expression of COX-2 and MMP-9 mRNA. Lactoferrin reduced LPS-induced cellular proliferation, inflammatory mediator gene expression, and prostaglandin E2 production by bovine PBMCs in vitro. These effects may be beneficial in reducing the impact of endotoxemia in neonates.

Emulsions Based on the Interactions Between Lactoferrin and Chitosans

In this work, purification of lactoferrin from whey was performed with high recovery rate. Lactoferrin was then exploited in the preparation of food emulsions. Two tertiary emulsions, formed by olive oil, lecithin, chitosan, and lactoferrin, were compared: both the emulsions showed similar turbidity and stability. In the secondary emulsion formed by oil/lecithin/chitosan, the pH was increased to 9 before addition of lactoferrin. Then, lactoferrin was added, and the pH was stabilized above pH 9. Lactoferrin was found in amounts of 1 to 2.5 mg/ml in the multiple experiments. A fraction of the added lactoferrin was also present in a milky layer above the emulsion layer. This was, to our knowledge, the first study of emulsions made exploiting the interactions between lactoferrin and chitosan. It was noted that chitosan droplets remained soluble, although the hydrocolloid solubility occurs at pH lower than 5.9. These results showed the feasibility of manufacturing lactoferrin-based emulsions as functional foods.

Formation of Multilayers at the Interface of Oil-in-Water Emulsion via Interactions between Lactoferrin and β-Lactoglobulin

The interactions between negatively charged β-lactoglobulin and the positively charged lactoferrin at the droplet surface to form a multi-protein surface layer were examined. Addition of lactoferrin to the aqueous phase of emulsions formed with β-lactoglobulin at pH 7.0 caused an increase in the ζ-potential of emulsion droplets, and the ζ-potential became positive as the concentration of added lactoferrin was higher than 1% in the system. It is found that lactoferrin binds to adsorbed β-lactoglobulin at droplet surface probably via electrostatic interactions. The amount of lactoferrin at interface increased with increasing the concentration of added lactoferrin, but it decreased with a decrease in the pH. No lactoferrin was observed at interface at pH 3 and 4. By contrast, when β-lactoglobulin was added in the emulsions formed with lactoferrin at pH 7.0, the ζ-potential of emulsions changed from positive to negative as the concentration of added β-lactoglobulin increased. The amount of β-lactoglobulin at surface increased correspondingly with increasing the concentration of added β-lactoglobulin. However, in this case, β-lactoglobulin remained bound at interface even at pH 3 and 4 where both lactoferrin and β-lactoglobulin are positively charged. The association of lactoferrin or β-lactoglobulin with the surface proteins that have oppositely charge is probably mainly through electrostatic interactions between the two proteins. It appears that alternative layers of these proteins could be created at the droplet surface.