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January 23, 2026Journal of Agricultural and Food Chemistry0 citationsOpen Access

Impact of Holder Pasteurization on Extracellular Vesicles and Immunoregulatory MicroRNAs in Human Breast Milk

CMC. MartínezPediatric Oncology Group LRLuis Javier Navarro RoyoUniversidad de Oviedo SCSara Escudero CernudaUniversidad de Oviedo

Key Points

The research aims to determine the effects of Holder pasteurization on extracellular vesicles and microRNA expression in human breast milk.
Evaluated the effects of Holder pasteurization at 62.5 °C for 30 min.
Analyzed extracellular vesicle morphology using transmission electron microscopy and dynamic light scattering.
Performed nanoparticle tracking analysis and protein quantification.
Measured miRNA expression using RT-qPCR on 13 selected miRNAs.
Stroke sizes of exosomes decreased after pasteurization, and protein concentration increased.
Total particle number of extracellular vesicles remained stable post-pasteurization.
Expression of eight out of 13 analyzed exosomal miRNAs significantly decreased.
MiRNA profiles in fat and serum fractions remained mostly unaffected.

Abstract

Human breast milk contains bioactive molecules, including microRNAs (miRNAs) that regulate neonatal development and immunity. Pasteurization ensures microbiological safety in milk banks, but its effects on milk-derived miRNAs remain unclear. This study evaluated the impact of Holder pasteurization (62.5 °C, 30 min) on extracellular vesicle (EVs) morphology and immunoregulatory miRNA expression in exosome, fat, and serum fractions from six donor samples. EVs were characterized by transmission electron microscopy, dynamic light scattering, nanoparticle tracking analysis, and protein quantification. Thirteen miRNAs were analyzed by RT-qPCR. Pasteurization caused morphological alterations in exosomes, reducing particle size and increasing protein concentration, while total particle number remained stable. Expression of eight out of 13 exosomal miRNAs significantly decreased (p ≤ 0.05), whereas miRNA profiles in fat and serum fractions were largely preserved.

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