Effects of Cow Milk-Derived Extracellular Vesicles on the Differentiation and Mineralization of Osteoblastic MC3T3-E1 Cells

Osteoporosis, which is caused by the structural weakness of bone tissue, is a systemic skeletal disease that decreases bone mass and increases the risk of fractures.For the aging population, osteoporosis is a major cause of morbidity and health expenditure.The objective of the present study was to investigate the potential anti-osteoporotic properties of cow milk-derived extracellular vesicles (EVs) on the differentiation and mineralization in osteoblastic MC3T3-E1 cells.MC3T3-E1 cells were cultured in 0, 1, 5, and 10 μg/mL cow milk-derived EVs for 3 and 7 days.It was observed that the average diameter of EVs from cow milk isolated by ultracentrifugation was 156±2.5 nm, and the presence of EV-associated marker protein CD63 and whey protein-specific marker lactoferrin was confirmed by immunoblot analysis.Our results determined that cow milk-derived EVs were increased osteoblastic cell proliferation and extracellular alkaline phosphatase (ALP) activity.Markedly, cow milk-derived EVs were significantly elevated the mineralized nodules in a does dependent manner for 3 and 7 days.Taken together, our results clearly demonstrated that cow milk-derived EVs may be useful in preventing osteoporosis by stimulating osteoblastic differentiation and mineralization.