Abstract：
Milk-derived extracellular vesicles (M-EVs) are low-cost, can be prepared in large quantities, and can cross the gastrointestinal barrier for oral administration. However, the composition of milk is complex, and M-EVs obtained by different extraction methods may affect their oral delivery. Based on this, a new method for extracting M-EVs based on cryogenic freezing treatment (Cryo-M-EVs) is proposed and compared with the previously reported acetic acid treatment (Acid-M-EVs) method and the conventional ultracentrifugation method (Ulltr-M-EVs). The new method simplifies the pretreatment step and achieves 25-fold and twofold higher yields than Acid-M-EVs and Ulltr-M-EVs. And it is interesting to note that Cryo-M-EVs and Acid-M-EVs have higher cellular uptake efficiency, and Cryo-M-EVs present the best transepithelial transport effect. After oral administration of the three M-EVs extracted by three methods in mice, Cryo-M-EVs effectively successfully cross the gastrointestinal barrier and achieve hepatic accumulation, whereas Acid-M-EVs and Ultr-M-EVs mostly reside in the intestine. The M-EVs obtained by the three extraction methods show a favorable safety profile at the cellular as well as animal level. Therefore, when M-EVs obtained by different extraction methods are used for oral drug delivery, their accumulation properties at different sites can be utilized to better deal with different diseases.
摘要：
牛奶来源的细胞外囊泡（M-EV）是低成本的，可以大量准备，并可通过胃肠屏障进行口服给药。然而,牛奶的成分很复杂，通过不同的提取方法获得的M-EV可能会影响其口服递送。基于此,我们提出了一种基于低温冷冻处理的提取M-EV的新方法（Cryo-M-EV），并将该方法与先前报道的乙酸处理（Acid-M-EV）方法和常规超速离心方法（Ulltr-M-EV）进行了比较。新方法简化了预处理步骤，并且比酸-M-EV和Ulltr-M-EV高25倍和2倍。有趣的是，Cryo-M-EV和Acid-M-EV具有更高的细胞摄取效率，和Cryo-M-EV表现出最佳的跨上皮运输效应。在小鼠中口服通过三种方法提取的三种M-EV后，Cryo-M-EV有效地穿过胃肠道屏障并实现肝脏蓄积，而酸-M-EVs和Ultr-M-EVs主要存在于肠中。通过三种提取方法获得的M-EV在细胞和动物水平上显示出有利的安全性。因此,当通过不同提取方法获得的M-EV用于口服药物递送时，我们可以利用它们在不同地点的积累特性来更好地应对不同的疾病。本文受版权保护。保留所有权利。
