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They reduce oxidative-stress marker expression, including DDIT4 and HIF1A.","Answer",{"name":66,"@type":61,"acceptedAnswer":67},"What treatment conditions and comparison groups were used in the study?",{"text":68,"@type":64},"BOECs were treated with mEVs or algae-derived EVs (aEVs, negative control) using three strategies: pre-treatment, co-incubation, and post-treatment. EVs were also compared across storage conditions (−80 °C storage vs lyophilization).",{"name":70,"@type":61,"acceptedAnswer":71},"Which components of mEVs are suggested to drive the protective effects?",{"text":72,"@type":64},"Enzyme digestion implies internal EV cargo contributes more than surface-associated molecules. 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Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.  \n1 Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, 51006 Tartu, Estonia  \n2 Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, Faculty of Medicine, University of Tartu, 14B Ravila, 50411 Tartu, Estonia  \n3 Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-748 Olsztyn, Poland  \n4 Institute of Chemistry, University of Tartu, 50411 Tartu, Estonia  \n5 Department of Animal Science, Faculty of Agriculture, University of Peradeniya, Peradeniya 20400, Sri Lanka  \n6 Division of Clinical Medicine, School of Medicine & Population Health, University of Sheffield, Sheffield S10 2SF, UK  \n* Correspondence: [alireza.fazeli@emu.ee](alireza.fazeli@emu.ee)  \nHighlights  \nWhat are the main findings?  \n• Milk extracellular vesicles (mEVs) attenuate CoCl 2-induced cytotoxicity and promote cell migration in bovine oviduct epithelial cells (BOECs) .  \n• Analysis of mEV miRNA and protein cargo revealed biological pathways that facilitate cellular recovery from oxidative stress (OS) .  \nWhat are the implications of the main findings?  \n• The protective effects of mEVs against CoCl2-induced stress in BOECs are likely mediated by their miRNA and protein cargo, providing a foundation for future mechanistic studies.  \n• mEVs may hold potential as a therapeutic approach for alleviating oviduct-associated OS.  \nAbstract  \nExtracellular vesicles (EVs) are promising therapeutic agents due to their role in intercellular communication. This study examined the protective effects of milk-derived EVs (mEVs) on bovine oviductal epithelial cells (BOECs) under cobalt chloride (CoCl 2)-induced oxidative stress (OS), comparing EVs stored at −80 ◦ C or lyophilized. mEVs and algae-derived EVs (aEVs; negative control) were isolated via tangential flow filtration and applied at 107, 109, and 1011 particles/mL in three treatment strategies: pre-treatment, co-incubation, and post-treatment. mEVs specifically enhanced cell viability in all protocols except for posttreatment, where only 107 particles/mL was effective; meanwhile, storage method did not affect EV activity. Enzyme digestion suggested that internal EV cargos are potentially the dominant contributors to the protective response compared to surface-associated molecules. mEVs reduced the expression of the OS markers DDIT4 and HIF1A while promoting cell migration more effectively than aEVs. Pathway enrichment analysis of previously reported mEV miRNAs indicated regulation of cytokine production and glucocorticoid responses, potentially contributing to OS defense. mEV protein cargo analysis showed pathways primarily linked to peptidase and vesicle-related functions, suggesting that protein cargo may also contribute to the observed protective effects. Overall, mEVs protect BOECs against CoCl 2-induced OS and maintain bioactivity after lyophilization.  \nKeywords: extracellular vesicles; oxidative stress; epithelial cell; miRNA  \n1. Introduction  \nThe oviduct is a crucial reproductive organ that links the ovary to the uterus and serves as the key site for transportation and the final maturation of gametes [1], fertilization, and early embryonic development in humans and other mammals [2–4] . The proper execution of these ","cbCaiswmqVT9rrRt","https://ap.wps.com/l/cbCaiswmqVT9rrRt","pdf",4093856,25,"English","# Highlights\n## Main findings\n## Implications\n# Abstract\n# Keywords\n# 1. Introduction\n## Oviduct physiology and ROS balance\n## Oxidative stress and reproductive disorders\n## Antioxidant interventions\n## Therapeutic potential of extracellular vesicles","[{\"question\":\"How do milk-derived extracellular vesicles (mEVs) affect oxidative stress in bovine oviduct epithelial cells?\",\"answer\":\"mEVs attenuate CoCl2-induced cytotoxicity and promote cell migration in BOECs. They reduce oxidative-stress marker expression, including DDIT4 and HIF1A.\"},{\"question\":\"What treatment conditions and comparison groups were used in the study?\",\"answer\":\"BOECs were treated with mEVs or algae-derived EVs (aEVs, negative control) using three strategies: pre-treatment, co-incubation, and post-treatment. EVs were also compared across storage conditions (−80 °C storage vs lyophilization).\"},{\"question\":\"Which components of mEVs are suggested to drive the protective effects?\",\"answer\":\"Enzyme digestion implies internal EV cargo contributes more than surface-associated molecules. Pathway enrichment of mEV miRNAs and protein cargo analysis indicate roles in cytokine/glucocorticoid-related regulation and peptidase/vesicle-related functions.\"}]","Milk-Derived Extracellular Vesicles Protect Bovine Oviduct Epithelial Cells from Oxidative Stress | PDF",1790769697,63]