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1
Hu, Yong, Johannes Thaler, and Rienk Nieuwland. "Extracellular Vesicles in Human Milk." Pharmaceuticals 14, no. 10 (2021): 1050. http://dx.doi.org/10.3390/ph14101050.
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Milk supports the growth and development of infants. An increasing number of mostly recent studies have demonstrated that milk contains a hitherto undescribed component called extracellular vesicles (EVs). This presents questions regarding why milk contains EVs and what their function is. Recently, we showed that EVs in human milk expose tissue factor, the protein that triggers coagulation or blood clotting, and that milk-derived EVs promote coagulation. Because bovine milk, which also contains EVs, completely lacks this coagulant activity, important differences are present in the biological functions of human milk-derived EVs between species. In this review, we will summarize the current knowledge regarding the presence and biochemical composition of milk EVs, their function(s) and potential clinical applications such as in probiotics, and the unique problems that milk EVs encounter in vivo, including survival of the gastrointestinal conditions encountered in the newborn. The main focus of this review will be human milk-derived EVs, but when available, we will also include information regarding non-human milk for comparison.
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Civra, Andrea, Rachele Francese, Manuela Donalisio, et al. "Human Colostrum and Derived Extracellular Vesicles Prevent Infection by Human Rotavirus and Respiratory Syncytial Virus in Vitro." Journal of Human Lactation 37, no. 1 (2021): 122–34. http://dx.doi.org/10.1177/0890334420988239.
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Background It is known that breastfeeding protects the infant from enteric and respiratory infections; however, the antiviral properties of human milk against enteric and respiratory viruses are largely unexplored. Research aims To explore the antiviral activity of human preterm colostrum against rotavirus and respiratory syncytial virus and to assess whether the derived extracellular vesicle contribute to this activity. Methods We used a cross-sectional, prospective two-group non-experimental design. Colostra were collected from mothers of preterm newborns ( N = 10) and extracellular vesicles were purified and characterized. The antiviral activity of colostra and derived extracellular vesicles were tested in vitro against rotavirus and respiratory syncytial virus and the step of viral replication inhibited by extracellular vesicles was investigated. Results Each sample of colostrum and colostrum-derived extracellular vesicles had significant antiviral activity with a wide interpersonal variability. Mechanism of action studies demonstrated that extracellular vesicles acted by interfering with the early steps of the viral replicative cycle. Conclusion We demonstrated the intrinsic antiviral activity of human colostrum against rotavirus and respiratory syncytial virus and we showed that extracellular vesicles substantially contribute to the overall protective effect. Our results contribute to unravelling novel mechanisms underlying the functional role of human milk as a protective and therapeutic agent in preterm infants.
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Prasadani, Madhusha, Suranga Kodithuwakku, Georgia Pennarossa, Alireza Fazeli, and Tiziana A. L. Brevini. "Therapeutic Potential of Bovine Milk-Derived Extracellular Vesicles." International Journal of Molecular Sciences 25, no. 10 (2024): 5543. http://dx.doi.org/10.3390/ijms25105543.
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Milk is a fundamental component of the human diet, owing to its substantial nutritional content. In addition, milk contains nanoparticles called extracellular vesicles (EVs), which have indicated their potential beneficial roles such as cell-to-cell communication, disease biomarkers, and therapeutics agents. Amidst other types of EVs, milk EVs (MEVs) have their significance due to their high abundance, easy access, and stability in harsh environmental conditions, such as low pH in the gut. There have been plenty of studies conducted to evaluate the therapeutic potential of bovine MEVs over the past few years, and attention has been given to their engineering for drug delivery and targeted therapy. However, there is a gap between the experimental findings available and clinical trials due to the many challenges related to EV isolation, cargo, and the uniformity of the material. This review aims to provide a comprehensive comparison of various techniques for the isolation of MEVs and offers a summary of the therapeutic potential of bovine MEVs described over the last decade, analyzing potential challenges and further applications. Although a number of aspects still need to be further elucidated, the available data point to the role of MEVs as a potential candidate with therapeutics potential, and the supplementation of MEVs would pave the way to understanding their in-depth effects.
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Prasadani, Madhusha, Suranga Kodithuwakku, GEORGIA PENNAROSSA, Alireza Fazeli, and Tiziana A.L. Brevini. "Therapeutic Potential of Bovine Milk-Derived Extracellular Vesicles." International Journal of Molecular Sciences 25, no. 10 (2024): 5543. https://doi.org/10.3390/ijms25105543.
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Milk is a fundamental component of the human diet, owing to its substantial nutritional content. In addition, milk contains nanoparticles called extracellular vesicles (EVs), which have indicated their potential beneficial roles such as cell-to-cell communication, disease biomarkers, and therapeutics agents. Amidst other types of EVs, milk EVs (MEVs) have their significance due to their high abundance, easy access, and stability in harsh environmental conditions, such as low pH in the gut. There have been plenty of studies conducted to evaluate the therapeutic potential of bovine MEVs over the past few years, and attention has been given to their engineering for drug delivery and targeted therapy. However, there is a gap between the experimental findings available and clinical trials due to the many challenges related to EV isolation, cargo, and the uniformity of the material. This review aims to provide a comprehensive comparison of various techniques for the isolation of MEVs and offers a summary of the therapeutic potential of bovine MEVs described over the last decade, analyzing potential challenges and further applications. Although a number of aspects still need to be further elucidated, the available data point to the role of MEVs as a potential candidate with therapeutics potential,
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Samuel, Monisha, Rahul Sanwlani, Mohashin Pathan, et al. "Isolation and Characterization of Cow-, Buffalo-, Sheep- and Goat-Milk-Derived Extracellular Vesicles." Cells 12, no. 20 (2023): 2491. http://dx.doi.org/10.3390/cells12202491.
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Milk is a complex biological fluid that has high-quality proteins including growth factors and also contains extracellular vesicles (EVs). EVs are a lipid bilayer containing vesicles that contain proteins, metabolites and nucleic acids. Several studies have proposed that EVs in cow milk can survive the gut and can illicit cross-species communication in the consuming host organism. In this study, we isolated and characterized extracellular vesicles from the raw milk of the four species of the Bovidae family, namely cow, sheep, goat and buffalo, that contribute 99% of the total milk consumed globally. A comparative proteomic analysis of these vesicles was performed to pinpoint their potential functional role in health and disease. Vesicles sourced from buffalo and cow milk were particularly enriched with proteins implicated in modulating the immune system. Furthermore, functional studies were performed to determine the anti-cancer effects of these vesicles. The data obtained revealed that buffalo-milk-derived EVs induced significantly higher cell death in colon cancer cells. Overall, the results from this study highlight the potent immunoregulatory and anti-cancer nature of EVs derived from the milk of Bovidae family members.
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Ryskaliyeva, A., Z. Krupova, C. Henry, B. Faye, G. Konuspayeva, and P. Martin. "Comprehensive proteomic analysis of camel milk-derived extracellular vesicles." International Journal of Biology and Chemistry 12, no. 2 (2019): 93–104. http://dx.doi.org/10.26577/ijbch-2019-i2-12.
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Galley, Jeffrey D., and Gail E. Besner. "The Therapeutic Potential of Breast Milk-Derived Extracellular Vesicles." Nutrients 12, no. 3 (2020): 745. http://dx.doi.org/10.3390/nu12030745.
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In the past few decades, interest in the therapeutic benefits of exosomes and extracellular vesicles (EVs) has grown exponentially. Exosomes/EVs are small particles which are produced and exocytosed by cells throughout the body. They are loaded with active regulatory and stimulatory molecules from the parent cell including miRNAs and enzymes, making them prime targets in therapeutics and diagnostics. Breast milk, known for years to have beneficial health effects, contains a population of EVs which may mediate its therapeutic effects. This review offers an update on the therapeutic potential of exosomes/EVs in disease, with a focus on EVs present in human breast milk and their remedial effect in the gastrointestinal disease necrotizing enterocolitis. Additionally, the relationship between EV miRNAs, health, and disease will be examined, along with the potential for EVs and their miRNAs to be engineered for targeted treatments.
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Somiya, Masaharu, Yusuke Yoshioka, and Takahiro Ochiya. "Biocompatibility of highly purified bovine milk-derived extracellular vesicles." Journal of Extracellular Vesicles 7, no. 1 (2018): 1440132. http://dx.doi.org/10.1080/20013078.2018.1440132.
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Hu, Yong, Lena Hell, Ruth Anna Kendlbacher, et al. "Human milk triggers coagulation via tissue factor–exposing extracellular vesicles." Blood Advances 4, no. 24 (2020): 6274–82. http://dx.doi.org/10.1182/bloodadvances.2020003012.
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Abstract Almost a century ago, it was discovered that human milk activates the coagulation system, but the milk component that triggers coagulation had until now been unidentified. In the present study, we identify this component and demonstrate that extracellular vesicles (EVs) present in normal human milk expose coagulant tissue factor (TF). This coagulant activity withstands digestive conditions, mimicking those of breastfed infants, but is sensitive to pasteurization of pooled donor milk, which is routinely used in neonatal intensive care units. In contrast to human milk, bovine milk, the basis of most infant formulas, lacks coagulant activity. Currently, the physiological function of TF-exposing vesicles in human milk is unknown, but we speculate that these vesicles may be protective for infants. Another explanation could be nipple skin damage, which occurs in most breastfeeding women. Milk-derived TF-exposing EVs may seal the wound and thereby reduce bleeding and breast inflammation.
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Ong, Siew Ling, Cherie Blenkiron, Stephen Haines, et al. "Ruminant Milk-Derived Extracellular Vesicles: A Nutritional and Therapeutic Opportunity?" Nutrients 13, no. 8 (2021): 2505. http://dx.doi.org/10.3390/nu13082505.
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Milk has been shown to contain a specific fraction of extracellular particles that are reported to resist digestion and are purposefully packaged with lipids, proteins, and nucleic acids to exert specific biological effects. These findings suggest that these particles may have a role in the quality of infant nutrition, particularly in the early phase of life when many of the foundations of an infant’s potential for health and overall wellness are established. However, much of the current research focuses on human or cow milk only, and there is a knowledge gap in how milk from other species, which may be more commonly consumed in different regions, could also have these reported biological effects. Our review provides a summary of the studies into the extracellular particle fraction of milk from a wider range of ruminants and pseudo-ruminants, focusing on how this fraction is isolated and characterised, the stability and uptake of the fraction, and the reported biological effects of these fractions in a range of model systems. As the individual composition of milk from different species is known to differ, we propose that the extracellular particle fraction of milk from non-traditional and minority species may also have important and distinct biological properties that warrant further study.
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Li, Ying, Xin Zhang, Chao Zhang, et al. "Comparative study on the immunomodulatory function of extracellular vesicles from different dairy products." Food & Function 13, no. 5 (2022): 2504–14. http://dx.doi.org/10.1039/d1fo02394b.
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Raghuwanshi, Gaurav, Harpritkaur Bagga, Gunjan Chaudhari, and Ujwal Vyas. "Milk-Derived Exosomes: Innovative Nanocarriers for Enhanced Anticancer Drug Delivery." INTERNATIONAL JOURNAL OF PHARMACEUTICAL QUALITY ASSURANCE 15, no. 03 (2024): 1767–77. http://dx.doi.org/10.25258/ijpqa.15.3.103.
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This in-depth review examines the recently emerging field of using exosomes from milk as anticancer medication nanocarriers. It gives a summary of the most current developments, difficulties, and opportunities in this cutting-edge therapeutic strategy. Exosomes are found in many different body fluids and are considered a promising option for precision medicine due to their biocompatibility and innate cell-targeting abilities. These extracellular vesicles are nanoscale. The review commences with a comprehensive synopsis of the composition and biogenesis of exosomes derived from milk, highlighting their distinct membrane properties and capacity to transport cargo. Interestingly, these naturally occurring nanocarriers hold a variety of bioactive substances that can be precisely delivered as anticancer medications. These substances include proteins, lipids, and nucleic acids. Because dietary(poly) phenols are rapidly metabolized, their ability to prevent cancer is limited. Extracellular vesicles called exosomes may shield polyphenols from metabolism. Our objective was to evaluate the anticancer effects of free curcumin and resveratrol in breast cancer cell lines compared to their encapsulation in extracellular matrix derived from milk. Breast tissue was disposed of kinetically using rats. Curcumin and resveratrol were assessed using UPLC-QTOF-MS and GV-MS, respectively. Dietary polyphenols have a limited capacity to prevent cancer due to their rapid metabolism. Extracellular vesicles called exosomes may shield polyphenols from metabolism. Our goal was to assess in breast tissue. UPLC-QTOF-MS and GV-MS were used to evaluate curcumin and resveratrol, respectively. Curcumin and Resveratrol anticancer activity and bioavailability were improved by milk extracellular urea, which served as Trojan horses to get around the ABC-mediated chemoresistance of cancer cells. Exosomes derived from milk are being studied for their potential as carriers of therapeutic and diagnostic agents, emphasizing the potential benefits of personalized and precision medicine approaches to cancer treatment. The review also covers the challenges that the clinical translation of milk-derived exosome-based drug delivery systems currently faces, including scalability, standardization and safety profiles. The article's conclusion presents an optimistic view of how milk-derived exosomes will develop in the future in terms of anticancer medication delivery. The review highlights the revolutionary potential of using milk-derived exosomes, nature's nanocarriers, to advance the field toward more precise and effective cancer treatments, anticipating future advancements and emerging trends.
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Sanwlani, Rahul, Pamali Fonseka, Sai V. Chitti, and Suresh Mathivanan. "Milk-Derived Extracellular Vesicles in Inter-Organism, Cross-Species Communication and Drug Delivery." Proteomes 8, no. 2 (2020): 11. http://dx.doi.org/10.3390/proteomes8020011.
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Milk is considered as more than a source of nutrition for infants and is a vector involved in the transfer of bioactive compounds and cells. Milk contains abundant quantities of extracellular vesicles (EVs) that may originate from multiple cellular sources. These nanosized vesicles have been well characterized and are known to carry a diverse cargo of proteins, nucleic acids, lipids and other biomolecules. Milk-derived EVs have been demonstrated to survive harsh and degrading conditions in gut, taken up by various cell types, cross biological barriers and reach peripheral tissues. The cargo carried by these dietary EVs has been suggested to have a role in cell growth, development, immune modulation and regulation. Hence, there is considerable interest in understanding the role of milk-derived EVs in mediating inter-organismal and cross-species communication. Furthermore, various attributes such as it being a natural source, as well as its abundance, scalability, economic viability and lack of unwarranted immunologic reactions, has generated significant interest in deploying milk-derived EVs for clinical applications such as drug delivery and disease therapy. In this review, the role of milk-derived EVs in inter-organismal, cross-species communication and in drug delivery is discussed.
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Zhong, Youxiu, Xudong Wang, Xian Zhao, et al. "Multifunctional Milk-Derived Small Extracellular Vesicles and Their Biomedical Applications." Pharmaceutics 15, no. 5 (2023): 1418. http://dx.doi.org/10.3390/pharmaceutics15051418.
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In recent years, small extracellular vesicles (sEVs) have been regarded as the next generation of novel delivery systems after lipid nanoparticles because of their advantages and huge prospects in drug delivery. Studies have shown that sEVs are abundant in milk and therefore can be a large and economical source of sEVs. Natural milk-derived small extracellular vesicles (msEVs) have important functions such as immune regulation, anti-bacterial infection, anti-oxidative, etc., and play a beneficial role in human health at multiple levels, including intestinal health, bone/muscle metabolism, and microbiota regulation. In addition, because they can pass the gastrointestinal barrier and have low immunogenicity, good biocompatibility, and stability, msEVs are considered a crucial oral drug delivery vehicle. Moreover, msEVs can be further engineered for targeted delivery to prolong the circulation time or enhance local drug concentrations. However, msEVs separation and purification, complex contents, and quality control hinder their application in drug delivery. This paper provides a comprehensive review of the biogenesis and characteristics, isolation and purification, composition, loading methods, and function of msEVs, based on which their applications in biomedical fields are further explored.
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Fonseka, Pamali, Taeyoung Kang, Sing Chee, et al. "Temporal Quantitative Proteomics Analysis of Neuroblastoma Cells Treated with Bovine Milk-Derived Extracellular Vesicles Highlights the Anti-Proliferative Properties of Milk-Derived Extracellular Vesicles." Cells 10, no. 4 (2021): 750. http://dx.doi.org/10.3390/cells10040750.
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Neuroblastoma (NBL) is a pediatric cancer that accounts for 15% of childhood cancer mortality. Amplification of the oncogene N-Myc occurs in 20% of NBL patients and is considered high risk as it correlates with aggressiveness, treatment resistance and poor prognosis. Even though the treatment strategies have improved in the recent years, the survival rate of high-risk NBL patients remain poor. Hence, it is crucial to explore new therapeutic avenues to sensitise NBL. Recently, bovine milk-derived extracellular vesicles (MEVs) have been proposed to contain anti-cancer properties. However, the impact of MEVs on NBL cells is not understood. In this study, we characterised MEVs using Western blotting, NTA and TEM. Importantly, treatment of NBL cells with MEVs decreased the proliferation and increased the sensitivity of NBL cells to doxorubicin. Temporal label-free quantitative proteomics of NBL cells highlighted the depletion of proteins involved in cell metabolism, cell growth and Wnt signalling upon treatment with MEVs. Furthermore, proteins implicated in cellular senescence and apoptosis were enriched in NBL cells treated with MEVs. For the first time, this study highlights the temporal proteomic profile that occurs in cancer cells upon MEVs treatment.
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Ascanius, Susanne R., Maria S. Hansen, Marie S. Ostenfeld та Jan T. Rasmussen. "Milk-Derived Extracellular Vesicles Suppress Inflammatory Cytokine Expression and Nuclear Factor-κB Activation in Lipopolysaccharide-Stimulated Macrophages". Dairy 2, № 2 (2021): 165–78. http://dx.doi.org/10.3390/dairy2020015.
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In milk and milk products, small membrane-enclosed vesicles can be found, commonly termed extracellular vesicles (EVs). Milk-derived EVs have previously been suggested to have immunoregulatory properties, especially important for infants without a fully functioning immune system. In the present study, EV fractions were isolated from human milk, mature and colostrum bovine milk, and two dairy fractions, and successively surveyed for their immunomodulating effects on lipopolysaccharide (LPS)-stimulated macrophages (RAW264.7). RAW264.7 cell material and supernatant were evaluated by monitoring degradation of IκBα in the NF-κB pathway, and IL-6 and IL-1β cytokine production, using Western blotting and enzyme-linked immunosorbent assaying, respectively. The results revealed that preincubation with EVs derived from raw human and bovine milk lowered the LPS-activated response of the NF-κB pathway. Additionally, it was found that preincubation with EVs, from human and bovine milk as well as dairy whey or skim milk-derived fractions, decreased secretion of proinflammatory cytokines from LPS-activated RAW264.7 cells. The findings that milk-derived EVs can change the inflammatory response in macrophages support the notion that milk EVs have an important role in mother-to-infant communication and protection of a newborn.
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Moccia, Valentina, Alessandro Sammarco, Laura Cavicchioli, Massimo Castagnaro, Laura Bongiovanni, and Valentina Zappulli. "Extracellular Vesicles in Veterinary Medicine." Animals 12, no. 19 (2022): 2716. http://dx.doi.org/10.3390/ani12192716.
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Extracellular vesicles (EVs) are cell-derived membrane-bound vesicles involved in many physiological and pathological processes not only in humans but also in all the organisms of the eukaryotic and prokaryotic kingdoms. EV shedding constitutes a fundamental universal mechanism of intra-kingdom and inter-kingdom intercellular communication. A tremendous increase of interest in EVs has therefore grown in the last decades, mainly in humans, but progressively also in animals, parasites, and bacteria. With the present review, we aim to summarize the current status of the EV research on domestic and wild animals, analyzing the content of scientific literature, including approximately 220 papers published between 1984 and 2021. Critical aspects evidenced through the veterinarian EV literature are discussed. Then, specific subsections describe details regarding EVs in physiology and pathophysiology, as biomarkers, and in therapy and vaccines. Further, the wide area of research related to animal milk-derived EVs is also presented in brief. The numerous studies on EVs related to parasites and parasitic diseases are excluded, deserving further specific attention. The literature shows that EVs are becoming increasingly addressed in veterinary studies and standardization in protocols and procedures is mandatory, as in human research, to maximize the knowledge and the possibility to exploit these naturally produced nanoparticles.
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Mecocci, Samanta, Massimo Trabalza-Marinucci, and Katia Cappelli. "Extracellular Vesicles from Animal Milk: Great Potentialities and Critical Issues." Animals 12, no. 23 (2022): 3231. http://dx.doi.org/10.3390/ani12233231.
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Other than representing the main source of nutrition for newborn mammals, milk delivers a sophisticated signaling system from mother to child that promotes postnatal health. The bioactive components transferred through the milk intake are important for the development of the newborn immune system and include oligosaccharides, lactoferrin, lysozyme, α-La, and immunoglobulins. In the last 15 years, a pivotal role in this mother-to-child exchange has been attributed to extracellular vesicles (EVs). EVs are micro- and nanosized structures enclosed in a phospholipidic double-layer membrane that are produced by all cell types and released in the extracellular environment, reaching both close and distant cells. EVs mediate the intercellular cross-talk from the producing to the receiving cell through the transfer of molecules contained within them such as proteins, antigens, lipids, metabolites, RNAs, and DNA fragments. The complex cargo can induce a wide range of functional modulations in the recipient cell (i.e., anti-inflammatory, immunomodulating, angiogenetic, and pro-regenerative modulations) depending on the type of producing cells and the stimuli that these cells receive. EVs can be recovered from every biological fluid, including blood, urine, bronchoalveolar lavage fluid, saliva, bile, and milk, which is one of the most promising scalable vesicle sources. This review aimed to present the state-of-the-art of animal-milk-derived EV (mEV) studies due to the exponential growth of this field. A focus on the beneficial potentialities for human health and the issues of studying vesicles from milk, particularly for the analytical methodologies applied, is reported.
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Chutipongtanate, Somchai, Supasek Kongsomros, Hatice Cetinkaya, et al. "Prenatal SARS-CoV-2 Infection Alters Human Milk-Derived Extracellular Vesicles." Cells 14, no. 4 (2025): 284. https://doi.org/10.3390/cells14040284.
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Human milk-derived extracellular vesicles (HMEVs) are key components in breast milk, promoting infant health and development. Maternal conditions could affect HMEV cargo; however, the impact of SARS-CoV-2 infection on HMEVs remains unknown. This study investigated the influence of SARS-CoV-2 infection during pregnancy on postpartum HMEV molecules. The median duration from SARS-CoV-2 test positivity to milk collection was 3 months. After defatting and casein micelle disaggregation, HMEVs were isolated from milk samples of nine mothers with prenatal SARS-CoV-2 and six controls by sequential centrifugation, ultrafiltration, and qEV-size exclusion chromatography. The presence of HMEV was confirmed via transmission electron microscopy. Nanoparticle tracking analysis demonstrated particle diameters of <200 nm and yields of >1 × 1011 particles per mL of milk. Western immunoblots detected ALIX, CD9, and HSP70, supporting the presence of HMEVs in the isolates. Cargo from thousands of HMEVs were analyzed using a multi-omics approach, including proteomics and microRNA sequencing, and predicted that mothers with prenatal SARS-CoV-2 infection produced HMEVs with enhanced functionalities involving metabolic reprogramming, mucosal tissue development, and immunomodulation. Our findings suggest that SARS-CoV-2 infection during pregnancy boosts mucosal site-specific functions of HMEVs, potentially protecting infants against viral infections. Further prospective studies should be pursued to reevaluate the short- and long-term benefits of breastfeeding in the post-COVID era.
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Rosa, Fernanda, Rafaela Santos, Tomas Lugo, Audrey Brown, and Ulrich Bickel. "Extracellular vesicles derived miRNAs from bovine milk modulate the inflammatory response to lipopolysaccharide in vitro." Journal of Immunology 210, no. 1_Supplement (2023): 167.38. http://dx.doi.org/10.4049/jimmunol.210.supp.167.38.
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Abstract Bovine milk is composed of a variety of biologically active compounds including mi-croRNAs (miRNA), which are found within extracellular vesicles (EVs). Extracellular vesicle-derived miRNAs may play a crucial role in the regulation of the immune response. Thus, this study aimed to identify specific miRNAs present in bovine milk that modulate the immune system’s response to infection. To address this, RAW-Blue™ Cells, a mouse macrophage reporter cell line was challenged with E. coli lipopolysaccharide (LPS) and incubated with a pool of miRNAs isolated from bovine milk through a 24-hour period. After incubation, QUANTI-Blue™ assay was used to detect the activation of the transcription factor NF-κB by the different treatments. Raw-Blue™ cells treated with miRNAs induced to some extend the signaling pathways leading to the activation of NF-κB relative to control cells (P&lt;0.01). Bovine-milk derived miRNAs suppressed LPS-induced NF-κB activity in a dose-dependent manner (P=0.01). To identify the top expressed miRNAs that are driving such effects in vitro, real-time qPCR assays were performed in the milk samples. The miRNA-profiling revealed the presence of miRNAs involved in tumor suppression and cell apoptosis. Overall, our preliminary findings indicated that miRNAs derived from bovine milk have the potential to modulate the immune response to LPS in vitro. Texas Tech University, School of Veterinary Medicine
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Zeng, Bin, Hailong Wang, Junyi Luo, et al. "Porcine Milk-Derived Small Extracellular Vesicles Promote Intestinal Immunoglobulin Production through pIgR." Animals 11, no. 6 (2021): 1522. http://dx.doi.org/10.3390/ani11061522.
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Secretory immunoglobulin A (SIgA) plays an important role in gut acquired immunity and mucosal homeostasis. Breast milk is the irreplaceable nutritional source for mammals after birth. Current studies have shown the potential functional role of milk-derived small extracellular vesicles (sEVs) and their RNAs cargo in intestinal health and immune regulation. However, there is a lack of studies to demonstrate how milk-derived sEVs affect intestinal immunity in recipient. In this study, through in vivo experiments, we found that porcine milk small extracellular vesicles (PM-sEVs) promoted intestinal SIgA levels, and increased the expression levels of polymeric immunoglobulin receptor (pIgR) both in mice and piglet. We examined the mechanism of how PM-sEVs increased the expression level of pIgR in vitro by using a porcine small intestine epithelial cell line (IPEC-J2). Through bioinformatics analysis, dual-luciferase reporter assays, and overexpression or knockdown of the corresponding non-coding RNAs, we identified circ-XPO4 in PM-sEVs as a crucial circRNA, which leads to the expression of pIgR via the suppression of miR-221-5p in intestinal cells. Importantly, we also observed that oral administration of PM-sEVs increased the level of circ-XPO4 and decreased the level of miR-221-5p in small intestine of piglets, indicating that circRNAs in milk-derived sEVs act as sponge for miRNAs in recipients. This study, for the first time, reveals that PM-sEVs have a capacity to stimulate intestinal SIgA production by delivering circRNAs to receptors and sponging the recipient’s original miRNAs, and also provides valuable data for insight into the role and mechanism of animal milk sEVs in intestinal immunity.
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García-Martínez, Jorge, Íñigo M. Pérez-Castillo, Rafael Salto, José M. López-Pedrosa, Ricardo Rueda, and María D. Girón. "Beneficial Effects of Bovine Milk Exosomes in Metabolic Interorgan Cross-Talk." Nutrients 14, no. 7 (2022): 1442. http://dx.doi.org/10.3390/nu14071442.
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Extracellular vesicles are membrane-enclosed secreted vesicles involved in cell-to-cell communication processes, identified in virtually all body fluids. Among extracellular vesicles, exosomes have gained increasing attention in recent years as they have unique biological origins and deliver different cargos, such as nucleic acids, proteins, and lipids, which might mediate various health processes. In particular, milk-derived exosomes are proposed as bioactive compounds of breast milk, which have been reported to resist gastric digestion and reach systemic circulation, thus being bioavailable after oral intake. In the present manuscript, we critically discuss the available evidence on the health benefits attributed to milk exosomes, and we provide an outlook for the potential future uses of these compounds. The use of milk exosomes as bioactive ingredients represents a novel avenue to explore in the context of human nutrition, and they might exert important beneficial effects at multiple levels, including but not limited to intestinal health, bone and muscle metabolism, immunity, modulation of the microbiota, growth, and development.
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Buratta, Sandra, Lorena Urbanelli, Alessia Tognoloni, et al. "Protein and Lipid Content of Milk Extracellular Vesicles: A Comparative Overview." Life 13, no. 2 (2023): 401. http://dx.doi.org/10.3390/life13020401.
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The characterization of the protein and lipid cargo of milk extracellular vesicles from different mammal species is crucial for understanding their biogenesis and biological functions, as well as for a comprehensive description of the nutritional aspects of animal milk for human diet. In fact, milk EVs have been reported to possess relevant biological effects, but the molecules/biochemical pathways underlying these effects have been poorly investigated. The biochemical characterization is an important initial step for the potential therapeutic and diagnostic use of natural or modified milk EVs. The number of studies analysing the protein and lipid composition of milk EVs is limited compared to that investigating the nucleic acid cargo. Here, we revised the literature regarding the protein and lipid content of milk EVs. Until now, most investigations have shown that the biochemical cargo of EVs is different with respect to that of other milk fractions. In addition, even if these studies derived mostly from bovine and human milk EVs, comparison between milk EVs from different animal species and milk EVs biochemical composition changes due to different factors including lactation stages and health status is also beginning to be reported.
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Weiskirchen, Ralf, Sarah K. Schröder, Sabine Weiskirchen, Eva Miriam Buhl, and Bodo Melnik. "Isolation of Bovine and Human Milk Extracellular Vesicles." Biomedicines 11, no. 10 (2023): 2715. http://dx.doi.org/10.3390/biomedicines11102715.
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Extracellular vesicles such as exosomes are small-sized, bilayered extracellular biovesicles generated by almost every cell and released into the surrounding body fluids upon the fusion of multivesicular bodies and the plasma membrane. Based on their origin, they are enriched with a variety of biologically active components including proteins, lipids, nucleic acids, cellular metabolites, and many other constituents. They can either attach or fuse with the membrane of a target cell, or alternatively be taking up via endocytosis by a recipient cell. In particular, milk exosomes have been recently shown to be a fundamental factor supporting infant growth, health, and development. In addition, exosomes derived from different cell types have been shown to possess regenerative, immunomodulatory, and anti-inflammatory properties, suggesting that they are a potential therapeutic tool in modulating the pathogenesis of diverse diseases. Therefore, efficient protocols for the isolation of milk exosomes in a high quantity and purity are the basis for establishing clinical applications. Here, we present an easy-to-follow protocol for exosome isolation from bovine and human milk. Electron microscopic analysis and nanoparticle tracking analysis reveal that the protocols allow the isolation of highly enriched fractions of exosomes. The purified exosomes express the typical exosomal protein markers, CD81 and ALIX.
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Seegobin, Nidhi, Marissa Taub, Cécile Vignal, et al. "Small milk-derived extracellular vesicles: Suitable vehicles for oral drug delivery?" European Journal of Pharmaceutics and Biopharmaceutics 212 (July 2025): 114744. https://doi.org/10.1016/j.ejpb.2025.114744.
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Babaker, Manal A., Fadwa A. Aljoud, Faris Alkhilaiwi, et al. "The Therapeutic Potential of Milk Extracellular Vesicles on Colorectal Cancer." International Journal of Molecular Sciences 23, no. 12 (2022): 6812. http://dx.doi.org/10.3390/ijms23126812.
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Colorectal cancer remains one of the leading prevalent cancers in the world and is the fourth most common cause of death from cancer. Unfortunately, the currently utilized chemotherapies fail in selectively targeting cancer cells and cause harm to healthy cells, which results in profound side effects. Researchers are focused on developing anti-cancer targeted medications, which is essential to making them safer, more effective, and more selective and to maximizing their therapeutic benefits. Milk-derived extracellular vesicles (EVs) from camels and cows have attracted much attention as a natural substitute product that effectively suppresses a wide range of tumor cells. This review sheds light on the biogenesis, methods of isolation, characterization, and molecular composition of milk EVs as well as the therapeutic potentials of milk EVs on colorectal cancer.
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Wang, Mengling, Meng Cai, Xiaoyan Zhu, Xuemei Nan, Benhai Xiong, and Liang Yang. "Comparative Proteomic Analysis of Milk-Derived Extracellular Vesicles from Dairy Cows with Clinical and Subclinical Mastitis." Animals 13, no. 1 (2023): 171. http://dx.doi.org/10.3390/ani13010171.
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Extracellular vesicles (EVs) are membranous vesicles found in biological fluids with essential functions. However, milk-derived EV proteins from clinical mastitis (CM) and subclinical mastitis (SM) cows have yet to be studied in detail. In this study, milk-derived EVs of CM, SM, and Healthy cows were extracted using a combination of acetic acid/ultracentrifugation and density gradient ultracentrifugation and analyzed using a shotgun proteomic by data-independent acquisition mode. A total of 1253 milk exosome proteins were identified and quantified. Differently enriched (DE) proteins were identified as given a Benjamini–Hochberg adjusted p < 0.05 and a fold change of at least 2. There were 53 and 1 DE proteins in milk-derived EVs from CM and SM cows compared with healthy cows. Protein S100-A9, Protein S100-A8, Chitinase-3-like protein 1, Haptoglobin, Integrin beta-2, and Chloride intracellular channel protein 1 were more abundant in the CM group (adjusted p < 0.05). Still, their enrichment in the SM group was not significant as in the Healthy group. The enrichment of DE proteins between CM and Healthy group was consistent with elevated GO (Gene Ontology) processes—defense response, defense response to Gram-positive bacterium, granulocyte chemotaxis also contributed to Reactome pathways—neutrophil degranulation, innate immune system, and antimicrobial peptides in the CM group. These results provide essential information on mastitis-associated proteins in milk-derived EVs and indicate the biological functions of milk-derived EVs proteins require further elucidation.
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Ou, Hairui, Tamas Imre Csuth, Tamas Czompoly, and Krisztian Kvell. "Dairy: Friend or Foe? Bovine Milk-Derived Extracellular Vesicles and Autoimmune Diseases." International Journal of Molecular Sciences 25, no. 21 (2024): 11499. http://dx.doi.org/10.3390/ijms252111499.
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Due to the availability, scalability, and low immunogenicity, bovine milk-derived extracellular vesicles (MEVs) are increasingly considered to be a promising carrier of nanomedicines for future therapy. However, considering that extracellular vesicles (EVs) are of biological origin, different sources of EVs, including the host origin and the specific cells that produce the EVs, may have different effects on the structure and function of EVs. Additionally, MEVs play an important role in immune regulation, due to their evolutionary conserved cargo, such as cytokines and miRNAs. Their potential effects on different organs, as well as their accumulation in the human body, should not be overlooked. In this review, we have summarized current impacts and research progress brought about by utilizing MEVs as nano-drug carriers. Nevertheless, we also aim to explore the possible connections between the molecules involved in cellular immunity, cytokines and miRNAs of MEVs produced under different health conditions, and autoimmune diseases.
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ROSARIO, CHARLUZ M. AROCHO, ADELA OLIVA CHAVEZ, CRAIG COATES, PETE TEEL, DONALD THOMAS, and ARTEM S. ROGOVSKYY. "Characterization and evaluation of extracellular vesicles as anti-tick vaccine candidates." Zoosymposia 22 (November 30, 2022): 315. http://dx.doi.org/10.11646/zoosymposia.22.1.197.
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Among all the agricultural sectors, the livestock industry is the most economically developed and prolific. Livestock production promotes economic stability globally and provides a significant source of protein in meat and milk. Ticks can transmit pathogens such as Anaplasma marginale, the causative agent of Bovine Anaplasmosis, affecting the cattle industry due to mortality, reduction of meat and milk production, and increased costs for treatment and prevention. Bovine Anaplasmosis results in estimated losses of over 300 million USD per year in the U.S. There are no effective treatments since they act as bacteriostatic antimicrobials, arresting bacterial growth, but do not eliminate them. In subclinical levels the infection can persist. Further, the ability of A. marginale to undergo antigenic variation hinders vaccine development. Therefore, alternatives that target transmission and early establishment of the pathogen are needed. Extracellular vesicles are small membrane blebs secreted by eukaryotic cells that act in cell-to-cell communication. These vesicles can be divided into exosomes and microvesicles, depending on their mechanism of biogenesis and secretion. Recent studies, and my preliminary data, have shown that tick extracellular vesicles carry proteomic material that is conserved among different tick species. This proteomic material can be used as a vaccine candidate for the development of anti-tick vaccines. We hypothesize that these vesicles have potential as vaccine candidates for tick control. To test this hypothesis, we will: 1) Characterize the vesicle populations secreted by Dermacentor andersoni, Amblyomma americanum, and Rhipicephalus microplus ticks, 2) Define the best adjuvant-vesicle combination to trigger strong humoral and cellular responses in cattle, and 3) Determine the effectiveness of extracellular vesicle-derived vaccines for the management of tick populations in semi-field conditions. The emergence of acaricidal resistance in tick populations increases the need to develop new alternative control methods. Current vaccines have shown to be protective only against particular tick populations and species, for this reason artificial vesicles containing conserved proteins among different tick species, present a tempting approach that can be mass-produced and commercialized as an anti-tick vaccine protective against different tick species but specially R. microplus. This technology can be applied to other systems and would have a great impact in animal and human health.
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del Pozo-Acebo, Lorena, M.-C. López de las Hazas, Joao Tomé-Carneiro, et al. "Bovine Milk-Derived Exosomes as a Drug Delivery Vehicle for miRNA-Based Therapy." International Journal of Molecular Sciences 22, no. 3 (2021): 1105. http://dx.doi.org/10.3390/ijms22031105.
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MicroRNAs (miRNAs) are small non-coding RNAs with a known role as mediators of gene expression in crucial biological processes, which converts them into high potential contenders in the ongoing search for effective therapeutic strategies. However, extracellular RNAs are unstable and rapidly degraded, reducing the possibility of successfully exerting a biological function in distant target cells. Strategies aimed at enhancing the therapeutic potential of miRNAs include the development of efficient, tissue-specific and nonimmunogenic delivery methods. Since miRNAs were discovered to be naturally transported within exosomes, a type of extracellular vesicle that confers protection against RNase degradation and increases miRNA stability have been proposed as ideal delivery vehicles for miRNA-based therapy. Although research in this field has grown rapidly in the last few years, a standard, reproducible and cost-effective protocol for exosome isolation and extracellular RNA delivery is lacking. We aimed to evaluate the use of milk-derived extracellular vesicles as vehicles for extracellular RNA drug delivery. With this purpose, exosomes were isolated from raw bovine milk, combining ultracentrifugation and size exclusion chromatography (SEC) methodology. Isolated exosomes were then loaded with exogenous hsa-miR148a-3p, a highly expressed miRNA in milk exosomes. The suitability of exosomes as delivery vehicles for extracellular RNAs was tested by evaluating the absorption of miR-148a-3p in hepatic (HepG2) and intestinal (Caco-2) cell lines. The potential exertion of a biological effect by miR-148a-3p was assessed by gene expression analysis, using microarrays. Results support that bovine milk is a cost-effective source of exosomes which can be used as nanocarriers of functional miRNAs with a potential use in RNA-based therapy. In addition, we show here that a combination of ultracentrifugation and SEC technics improve exosome enrichment, purity, and integrity for subsequent use.
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Sprenger, Richard R., Marie S. Ostenfeld, Ann Bjørnshave, Jan T. Rasmussen, and Christer S. Ejsing. "Lipidomic Characterization of Whey Concentrates Rich in Milk Fat Globule Membranes and Extracellular Vesicles." Biomolecules 14, no. 1 (2023): 55. http://dx.doi.org/10.3390/biom14010055.
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Lipids from milk fat globule membranes (MFGMs) and extracellular vesicles (EVs) are considered beneficial for cognitive development and human health. Milk-derived whey concentrates rich in these lipids are therefore used as ingredients in infant formulas to mimic human milk and in medical nutrition products to improve the metabolic fitness of adults and elderly people. In spite of this, there is no consensus resource detailing the multitude of lipid molecules in whey concentrates. To bridge this knowledge gap, we report a comprehensive and quantitative lipidomic resource of different whey concentrates. In-depth lipidomic analysis of acid, sweet, and buttermilk whey concentrates identified 5714 lipid molecules belonging to 23 lipid classes. The data show that the buttermilk whey concentrate has the highest level of fat globule-derived triacylglycerols and that the acid and sweet whey concentrates have the highest proportions of MFGM- and EV-derived membrane lipids. Interestingly, the acid whey concentrate has a higher level of cholesterol whereas sweet whey concentrate has higher levels of lactosylceramides. Altogether, we report a detailed lipid molecular compendium of whey concentrates and lay the groundwork for using in-depth lipidomic technology to profile the nutritional value of milk products and functional foods containing dairy-based concentrates.
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Ukkola, Jonne, Feby W. Pratiwi, Santeri Kankaanpää, et al. "Enrichment of bovine milk-derived extracellular vesicles using surface-functionalized cellulose nanofibers." Carbohydrate Polymers 297 (December 2022): 120069. http://dx.doi.org/10.1016/j.carbpol.2022.120069.
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Zhang, Sitong, Xipeng Sun, Yameng Zhang, et al. "Metabolomics analysis of extracellular vesicles derived from bovine colostrum and mature milk." Food Science of Animal Products 2, no. 3 (2024): 9240078. http://dx.doi.org/10.26599/fsap.2024.9240078.
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Reif, Shimon, Liron Birimberg-Schwartz, Myriam Grunewald, et al. "The Effect of Milk-Derived Extracellular Vesicles on Intestinal Epithelial Cell Proliferation." International Journal of Molecular Sciences 25, no. 24 (2024): 13519. https://doi.org/10.3390/ijms252413519.
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Inflammatory bowel disease (IBD) is a chronic, relapsing inflammation disorder of the gastrointestinal tract characterized by disrupted intestinal epithelial barrier function. Despite advances in treatment, including biological agents, achieving sustained remission remains challenging for many patients with IBD. This highlights the urgent need for novel therapeutic strategies. Milk-derived extracellular vesicles (MDEs) have emerged as a promising therapeutic option. In this study, we isolated and characterized MDEs and evaluated their effects on the function of intestinal epithelial cells (IECs). Using a murine model of Dextran Sulfate Sodium (DSS)-induced colitis, we observed that MDEs significantly ameliorated disease symptoms. The upregulation of β-catenin, a crucial mediator of Wnt signaling, in colonic tissues suggests that MDEs may facilitate epithelial regeneration and restore barrier function. In patient-derived colon organoids (PDCOs), MDEs were internalized and modulated the expression of key signaling molecules, such as the upregulation of β-catenin, cyclin D1, and the proliferation marker Ki67, indicating their potential to promote IEC proliferation and intestinal barrier repair. Importantly, MDEs demonstrated selective activity by downregulating β-catenin and cyclin D1 in colon cancer cells, leading to reduced proliferation. This selectivity indicates a dual therapeutic potential of MDEs for promoting healthy IEC proliferation while potentially mitigating malignancy risks.
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Jang, Hochung, and Yoosoo Yang. "Harnessing Milk-derived Extracellular Vesicles for Oral Drug Delivery and Therapeutic Application." Journal of Digestive Cancer Research 13, no. 1 (2025): 30–37. https://doi.org/10.52927/jdcr.2025.13.1.30.
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Carobolante, Greta, Julia Mantaj, Enrico Ferrari, and Driton Vllasaliu. "Cow Milk and Intestinal Epithelial Cell-Derived Extracellular Vesicles as Systems for Enhancing Oral Drug Delivery." Pharmaceutics 12, no. 3 (2020): 226. http://dx.doi.org/10.3390/pharmaceutics12030226.
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Ingestion is the preferred way for drug administration. However, many drugs have poor oral bioavailability, warranting the use of injections. Extracellular vesicles (EVs) from cow milk have shown potential utility in improving oral drug bioavailability. However, EVs produced by intestinal epithelial cells have not been investigated for this application. We compared the capacity of cow milk EVs and intestinal epithelial cell-derived counterparts to enhance oral drug bioavailability. EVs were isolated, fluorescently labelled, and loaded with curcumin (CUR) as a model poorly absorbable drug. These were then characterised before testing in an intestinal model (Caco-2). Epithelial cell-derived EVs showed notably higher cell uptake compared to cow milk EVs. Cell uptake was significantly higher in differentiated compared to undifferentiated cells for both types of EVs. While both milk- and cell-derived EVs improved the cell uptake and intestinal permeability of CUR (confirming oral drug bioavailability enhancement potential), epithelial cell EVs demonstrated a superior effect.
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Pollott, Geoff, Amanda Brito, Christopher Gardiner, and Charlotte Lawson. "A Comparison of Different Methodologies for the Measurement of Extracellular Vesicles and Milk-derived Particles in Raw Milk from Cows." Biomarker Insights 11 (January 2016): BMI.S38438. http://dx.doi.org/10.4137/bmi.s38438.
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Cow's milk is economically important to the agricultural industry with the nutritive value of milk being routinely measured. This does not give full insight into normal mammary tissue turnover during the course of lactation, which could be important for both an understanding of milk production and animal welfare. We have previously demonstrated that submicron particles, including extracellular vesicles (EVs), can be measured in unprocessed cow's milk by flow cytometry and that they correlate with stage of lactation. A number of different techniques are available to measure EVs and other milk-derived particles. The purpose of this study was to compare two different methodologies and the value of fluorescent staining for the phospholipid phosphatidylserine (PS), which is exposed on the surface of EVs (but not other milk-derived particles). We used two different flow cytometers and nanotracker analysis to detect milk-derived particles in whole and skimmed milk samples. Our findings indicate significant correlation, after staining for PS, suggesting potential for larger multicenter studies in the future.
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Benedetto, Alessandro, Nunzia Giaccio, Maddalena Arigoni, et al. "miRNome Characterization of Milk-Derived Extracellular Vesicles in Recombinant Somatotropin-Treated Dairy Cows." International Journal of Molecular Sciences 26, no. 6 (2025): 2437. https://doi.org/10.3390/ijms26062437.
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The recombinant bovine somatotropin (rbST) is a synthetic hormone developed to mimic the effects of the endogenous growth hormone, also known as bovine somatotropin (bST). Although rbST use in dairy cows is authorized in several countries, it is currently banned in Europe. Different methods for screening and confirmatory detection of rbST were developed, mainly based on LC-MS/MS and immune-enzymatic assays. However, some commercial forms of rbST have above the same amino acid sequence of bST, making it difficult to produce a reliable differentiation of recombinant from endogenous forms. Complementary strategies for indirect detection of rbST can therefore be considered as alternative biomarker-based tools. Untargeted transcriptomics was applied to characterize the microRNAs (miRNA) isolated from milk extracellular vesicles (EVs) in rbST-treated animals, aiming the identification of non-coding biomarkers related to its administration. Sequencing analysis of 63 archive samples collected during previous animal trial allowed for the identification of 35 differentially expressed (DE) miRNAs. A validation study performed by qPCR on a further 70 milk samples from a field survey confirmed the significant upregulation of bta-miR-10167-3p in milk EV from rbST-treated cows. The results obtained suggest the potential use of bta-miR-10167-3p as a non-invasive biomarker to be considered in novel screening strategies, needed to tackle rbST misuse in dairy cows.
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Macia, Laurence, Ralph Nanan, Elham Hosseini-Beheshti, and Georges E. Grau. "Host- and Microbiota-Derived Extracellular Vesicles, Immune Function, and Disease Development." International Journal of Molecular Sciences 21, no. 1 (2019): 107. http://dx.doi.org/10.3390/ijms21010107.
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Extracellular vesicles (EVs) are blebs of either plasma membrane or intracellular membranes carrying a cargo of proteins, nucleic acids, and lipids. EVs are produced by eukaryotic cells both under physiological and pathological conditions. Genetic and environmental factors (diet, stress, etc.) affecting EV cargo, regulating EV release, and consequences on immunity will be covered. EVs are found in virtually all body fluids such as plasma, saliva, amniotic fluid, and breast milk, suggesting key roles in immune development and function at different life stages from in utero to aging. These will be reviewed here. Under pathological conditions, plasma EV levels are increased and exacerbate immune activation and inflammatory reaction. Sources of EV, cells targeted, and consequences on immune function and disease development will be discussed. Both pathogenic and commensal bacteria release EV, which are classified as outer membrane vesicles when released by Gram-negative bacteria or as membrane vesicles when released by Gram-positive bacteria. Bacteria derived EVs can affect host immunity with pathogenic bacteria derived EVs having pro-inflammatory effects of host immune cells while probiotic derived EVs mostly shape the immune response towards tolerance.
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Abou el qassim, Loubna, Regina Golan-Gerstl, Shimon Reif, and Luis J. Royo. "Association between Dairy Production System and Milk Functionality Based on Analysis of miRNAs in Exosomes from Milk." Animals 14, no. 20 (2024): 2960. http://dx.doi.org/10.3390/ani14202960.
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Dairy farming practices significantly affect the nutritional and functional properties of milk. This study compares miRNAs in milk exosomes from extensive and intensive dairy systems and explores their potential implications for human consumers. Extensive systems are believed to produce milk of higher quality with better animal welfare compared to intensive systems. Milk samples from eight extensive and nine intensive dairy farms were analysed. Milk-derived extracellular vesicles were isolated through sequential ultracentrifugation and characterised through Dynamic Light Scattering and Nanosight to determine the size and the concentration of the extracellular vesicles, in addition to immunoblotting to ensure the presence of exosome-specific proteins in their membrane. miRNA levels were quantified using RT-qPCR, and metabolic pathways associated with miRNAs showing significant differences between farm groups were analysed. EVs from extensive farms had higher concentrations. Notably, bta-miR-451 levels were significantly higher in milk from extensive farms (p = 0.021). Like human miRNA hsa-miR-451, it is linked to pathways related to Parkinson’s disease and cancer. Our research suggests that milk production in extensive systems not only provides socioeconomic and environmental benefits but may also have positive effects on human health. Further research is warranted to explore the bioactive potential of these miRNAs and their implications for human health.
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Mun, Daye, Sangnam Oh, and Younghoon Kim. "Perspectives on Bovine Milk-Derived Extracellular Vesicles for Therapeutic Applications in Gut Health." Food Science of Animal Resources 42, no. 2 (2022): 197–209. http://dx.doi.org/10.5851/kosfa.2022.e8.
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Pisano, Courtney, Jeffrey Galley, Mostafa Elbahrawy, et al. "Human Breast Milk-Derived Extracellular Vesicles in the Protection Against Experimental Necrotizing Enterocolitis." Journal of Pediatric Surgery 55, no. 1 (2020): 54–58. http://dx.doi.org/10.1016/j.jpedsurg.2019.09.052.
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Liang, Jia Qi, Mei-Ying Xie, Lian-Jie Hou, et al. "miRNAs derived from milk small extracellular vesicles inhibit porcine epidemic diarrhea virus infection." Antiviral Research 212 (April 2023): 105579. http://dx.doi.org/10.1016/j.antiviral.2023.105579.
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Liu, Runyuan, Shuo Liu, Saixuan Wu, et al. "Milk-Derived Small Extracellular Vesicles Promote Osteogenic Differentiation and Inhibit Inflammation via microRNA-21." International Journal of Molecular Sciences 24, no. 18 (2023): 13873. http://dx.doi.org/10.3390/ijms241813873.
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Chronic apical periodontitis (CAP) is a disease with characteristics of inflammation and bone loss. In this study, our objective was to examine the function of small extracellular vesicles (sEVs) obtained from milk in encouraging osteogenic differentiation and inhibiting inflammation by miR-21 in CAP. The expression of miR-21 was detected using qRT-PCR in human CAP samples. The impact of miR-21 on the process of osteogenic differentiation was investigated using CCK-8, qRT-PCR, immunofluorescence staining, and Western blot analysis. The evaluation of RAW 264.7 cell polarization and the assessment of inflammatory factor expression were conducted through qRT-PCR. The influence of sEVs on MC3T3-E1 cells and RAW 264.7 cells was examined, with a particular emphasis on the involvement of miR-21. In human CAP samples, a decrease in miR-21 expression was observed. MiR-21 increased the expression of osteogenesis-related genes and M2 polarization genes while decreasing the expression of M1 polarization genes and inflammatory cytokines. Treatment with milk-derived sEVs also promoted osteogenesis and M2 polarization while inhibiting M1 polarization and inflammation. Conversely, the addition of miR-21 inhibitors resulted in opposite effects. Our results indicated that sEVs derived from milk had a positive effect on bone formation and activation of anti-inflammatory (M2) macrophages and simultaneously reduced inflammation by regulating miR-21 in CAP.
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Du, Chunmei, Kun Wang, Yiguang Zhao, et al. "Supplementation with Milk-Derived Extracellular Vesicles Shapes the Gut Microbiota and Regulates the Transcriptomic Landscape in Experimental Colitis." Nutrients 14, no. 9 (2022): 1808. http://dx.doi.org/10.3390/nu14091808.
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Harboring various proteins, lipids, and RNAs, the extracellular vesicles (EVs) in milk exert vital tissue-specific immune-protective functions in neonates via these bioactive cargos. This study aims to explore the anti-inflammatory effects of bovine milk-derived EVs on a dextran sulfate sodium (DSS)-induced colitis model and to determine the underlying molecular mechanisms. Sixty C57BL/6 mice were divided into the NC group (normal control), DSS group (DSS + PBS), DSS + LOW group (DSS + 1.5 × 108 p/g EVs), DSS + MID group (DSS + 1.5 × 109 p/g EVs), and DSS + HIG group (DSS + 1.0 × 1010 p/g EVs). Histopathological sections, the gut microbiota, and intestinal tissue RNA-Seq were used to comprehensively evaluate the beneficial functions in mitigating colitis. The morphology exhibited that the milk-derived EVs contributed to the integrity of the superficial epithelial structure in the intestine. Additionally, the concentrations of IL-6 and TNF-α in the colon tissues were significantly decreased in the EVs-treated mice. The abundances of the Dubosiella, Bifidobacterium, UCG-007, Lachnoclostridium, and Lachnospiraceae genera were increased in the gut after treatment with the milk-derived EVs. Additionally, the butyrate and acetate production were enriched in feces. In addition, 1659 genes were significantly down-regulated and 1981 genes were significantly up-regulated in the EVs-treated group. Meanwhile, 82 lncRNAs and 6 circRNAs were also differentially expressed. Overall, the milk-derived EVs could attenuate colitis through optimizing gut microbiota abundance and by manipulating intestinal gene expression, implying their application potential for colitis prevention.
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Benmoussa, Abderrahim, Jonathan Laugier, Charles Joly Beauparlant, Marine Lambert, Arnaud Droit, and Patrick Provost. "Complexity of the microRNA transcriptome of cow milk and milk-derived extracellular vesicles isolated via differential ultracentrifugation." Journal of Dairy Science 103, no. 1 (2020): 16–29. http://dx.doi.org/10.3168/jds.2019-16880.
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Ylioja, Caroline M., Laman K. Mamedova, and Barry J. Bradford. "332 Young Scholar Presentation: regulation of immune signaling by extracellular vesicles." Journal of Animal Science 97, Supplement_2 (2019): 132–33. http://dx.doi.org/10.1093/jas/skz122.235.
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Abstract Exosomes and other extracellular vesicles traffic a variety of protein and nucleic acid cargo throughout the body. Immune cells especially utilize these vesicles for communication, and they are being studied in clinical settings for their potential to act as disease markers, but also to serve as drug delivery systems. In animal science, a large part of the research around exosomes has centered around their presence in milk, as well as their bioavailability and potential impact for the neonate and for humans; milk-derived exosomes are enriched with immune-related proteins and microRNA that may relate to maternal health, mammary function, or neonate development. We sequenced miRNA isolated from colostrum of dairy cows predicted to feature either moderate or high degree of immune suppression, but found minimal differences (2 differentially expressed miRNA of 343 miRNA analyzed) between the two groups. Immune suppression exhibited during early lactation may also be related to altered exosome-mediated communication between immune cells. We studied the ability of bovine exosomes to alter immune responses of primary bovine monocyte-derived macrophages and found that exosomes alone or in combination with LPS were able to stimulate cytokine production to a greater extent than LPS alone. The ability of exosomes to transport cytokine, bioactive lipid, and regulatory RNA cargo suggests potential involvement in transition cow health. Altered exosome content and function have been associated with uterine infection, mastitis, and metabolic dysfunction; future studies with exosome signaling may help clarify the complexities of transition cow immune function and point to strategies to support immunity.
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Fan, Yuqin, Zhikang Li, Yanmei Hou, et al. "Effects of Different Processing on miRNA and Protein in Small Extracellular Vesicles of Goat Dairy Products." Nutrients 16, no. 24 (2024): 4331. https://doi.org/10.3390/nu16244331.
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Objectives: Small extracellular vesicles (sEVs) are nanosized vesicles with biological activities that exist in milk, playing functional roles in immunity, gut balance, and the nervous system. Currently, little is known about the impact of processing on milk sEVs. Methods: In this study, sEVs were collected from raw goat milk (g-sEV), pasteurized goat milk (pg-sEV), and goat milk powder (p-sEV) using a sucrose cushion centrifugation combined with qEV chromatography. Then, the sEVs were identified and compared using NTA, Western blot, and TEM. After extracting RNA and the total proteome from sEVs derived from different samples, the RNA was subjected to high-throughput sequencing, and peptide fragments were analyzed using mass spectrometry. Finally, GO and KEGG pathway analyses were performed on the results. Results: The characterization results revealed a decrease in diameter as the level of processing increased. High-throughput sequencing results showed that all three types of small extracellular vesicles were found to be rich in miRNA, and no significant differences were observed in the most abundant sEV species. Comparing with g-sEV, there were 3938 and 4645 differentially expressed miRNAs in pg-sEV and p-sEV, respectively, with the majority of them (3837 and 3635) being downregulated. These differentially expressed miRNAs were found to affect biological processes or signaling pathways such as neurodevelopment, embryonic development, and transcription. Proteomic analysis showed that there were 339 differentially expressed proteins between g-sEV and pg-sEV, with 209 proteins being downregulated. Additionally, there were 425 differentially expressed proteins between g-sEV and p-sEV, with 293 proteins being downregulated. However, no significant differences were observed in the most abundant protein species among the three types of sEVs. Enrichment analysis indicated that the differentially expressed proteins were associated with inflammation, immunity, and other related processes. Conclusions: These results indicate that extracellular vesicles have a protective effect on their cargo, while processing steps can have an impact on the size and quantity of the sEVs. Furthermore, processing can also lead to the loss of immune-related miRNA and proteins in sEVs.
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Barathan, Muttiah, Sook Luan Ng, Yogeswaran Lokanathan, Min Hwei Ng, and Jia Xian Law. "Milk-Derived Extracellular Vesicles: A Novel Perspective on Comparative Therapeutics and Targeted Nanocarrier Application." Vaccines 12, no. 11 (2024): 1282. http://dx.doi.org/10.3390/vaccines12111282.
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Milk-derived extracellular vesicles (mEVs) are emerging as promising therapeutic candidates due to their unique properties and versatile functions. These vesicles play a crucial role in immunomodulation by influencing macrophage differentiation and cytokine production, potentially aiding in the treatment of conditions such as bone loss, fibrosis, and cancer. mEVs also have the capacity to modulate gut microbiota composition, which may alleviate the symptoms of inflammatory bowel diseases and promote intestinal barrier integrity. Their potential as drug delivery vehicles is significant, enhancing the stability, solubility, and bioavailability of anticancer agents while supporting wound healing and reducing inflammation. Additionally, bovine mEVs exhibit anti-aging properties and protect skin cells from UV damage. As vaccine platforms, mEVs offer advantages including biocompatibility, antigen protection, and the ability to elicit robust immune responses through targeted delivery to specific immune cells. Despite these promising applications, challenges persist, including their complex roles in cancer, effective antigen loading, regulatory hurdles, and the need for standardized production methods. Achieving high targeting specificity and understanding the long-term effects of mEV-based therapies are essential for clinical translation. Ongoing research aims to optimize mEV production methods, enhance targeting capabilities, and conduct rigorous preclinical and clinical studies. By addressing these challenges, mEVs hold the potential to revolutionize vaccine development and targeted drug delivery, ultimately improving therapeutic outcomes across various medical fields.
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Arntz, Onno J., Bartijn C. H. Pieters, Marina C. Oliveira, et al. "Oral administration of bovine milk derived extracellular vesicles attenuates arthritis in two mouse models." Molecular Nutrition & Food Research 59, no. 9 (2015): 1701–12. http://dx.doi.org/10.1002/mnfr.201500222.
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