Milk extracellular vesicles (mEVs) derived from colostrum (C-mEVs) and mature milk (M-mEVs) have been proposed as oral drug delivery platforms and potential nanomedicines. However, their differences in oral administration properties and physiological functions have not yet been documented. Here, we comprehensively evaluated the biological properties and physiological functions of C-mEVs and M-mEVs, emphasizing their storage stability, gastrointestinal tolerability, and functional differences revealed through small RNA sequencing. Our findings showed that both C-mEVs and M-mEVs exhibit excellent stability at 4 and –80 ℃ over 1 month. Furthermore, C-mEVs, characterized by their smaller size, demonstrated greater tolerance to harsh digestive environments compared to M-mEVs. For physiological functions, both C-mEVs and M-mEVs suppressed inflammation, protected epithelial barrier integrity, and mitigated oxidative stress damage. Notably, M-mEVs displayed enhanced anti-inflammatory and antioxidative effects. Taken together, M-mEVs hold potential as nanotherapeutic agents for immunoinflammatory diseases, while C-mEVs may serve as a promising oral drug delivery platform.

Background: We aim to develop and validate the empirical dietary inflammatory index for pregnant women in the second trimester (EDII-PW) and assess its predictive value for maternal and neonatal outcomes. Methods: We utilized data from three sub-cohorts conducted in Hefei, China, including the Maternal and Infant Health Cohort (MIH-I: 2015-2019) with 5,052 pregnant women, MIH-II (2018-2022) with 1,932 pregnant women, and MIH-III (2023-2024) with 1,225 pregnant women, all recruited between gestational ages of 16 and 23 weeks. The development and validation of the EDII-PW index were based on three plasma inflammatory biomarkers (IL-6, hsCRP, and TNF-α) in MIH-I. In MIH-II, three additional biomarkers (E-selectin, VCAM-1, and ICAM-1) were included for independent biomarker validation. In MIH-III, six biomarkers (hsCRP, IL-6, TNF-α, E-selectin, VCAM-1, and ICAM-1) were measured in 1,225 participants and were used to derive inflammatory and endothelial biomarker profiles for the outcome prediction analyses. Results: The EDII-PW was significantly associated with plasma concentrations of inflammatory biomarkers. In MIH-I, participants in the highest quartile of EDII-PW had significantly higher levels of hsCRP, IL-6, and TNF-α compared to those in the lowest quartile. Similar associations were observed for E-selectin, VCAM-1, and ICAM-1 in MIH-II. In MIH-III, ROC curve analyses suggested that multivariable models containing the EDII-PW may discriminate maternal and neonatal outcomes, with AUC values as follows: 0.81 for premature rupture of membranes (PROM), 0.83 for gestational diabetes mellitus (GDM), 0.74 for endothelial dysfunction, 0.81 for inflammatory status, 0.80 for low birth weight (LBW), 0.82 for small for gestational age (SGA), 0.83 for preterm birth, and 0.76 for gestational hypertension. Conclusion: The EDII-PW showed reproducible associations with inflammatory biomarkers across independent sub-cohorts and potential utility as a component of multivariable models for adverse maternal and neonatal outcomes.

Milk-derived extracellular vesicles (mEVs) derived from bovine colostrum (C-mEVs) and mature milk (M-mEVs) are known to carry bioactive molecules with significant immunomodulatory properties and biological effects. However, most research on mEVs has focused on their proteomics, lipidomics, and transcriptome, while metabolome analysis are still lacking. This study aims to explore the metabolomic profiles of C-mEVs and M-mEVs to uncover their molecular characteristics and potential biological functions. The mEVs were purified from both bovine colostrum and mature milk using ultracentrifugation combined with size exclusion chromatography. Through untargeted metabolomic analysis employing liquid chromatography-tandem mass spectrometry (LC-MS/MS), 76 differential metabolites, such as leucylproline, levonordefrin, adenine, polyglycerol esters of fatty acids, and hypoxanthine were identified. Among them, lipid-related metabolites were the most prominent. Hierarchical cluster analysis and principal component analysis revealed distinct metabolic signatures between C-mEVs and M-mEVs. Pathway enrichment analysis indicated that the metabolites were involved in purine metabolism and ABC transporter pathways, which was closely related to cancer, inflammation, gastrointestinal disorders, and metabolic diseases. The findings highlight the potential of metabolomics to provide a temporally precise and detailed snapshot of the molecular properties of mEVs, contributing valuable insights into mEVs-mediated molecular mechanisms and offering new avenues for research into their biological effects and therapeutic applications.