Transcytosis of Bacillus subtilis extracellular vesicles through an in vitro intestinal epithelial cell model.

Ana Paula Domínguez Rubio, Jimena Martínez, Marcos Palavecino, Federico Fuentes, Christian Miquel Sánchez López, Antonio Marcilla, Oscar Edgardo Pérez, Mariana Piuri

Journal: Scientific reports 2020;10(1):3120

PMID: 32080346

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Abstract

["Bacterial EVs have been related to inter-kingdom communication between probiotic\/pathogenic bacteria and their hosts. Our aim was to investigate the transcytosis process of B. subtilis EVs using an in vitro intestinal epithelial cell model. In this study, using Confocal Laser Scanning Microscopy, we report that uptake and internalization of CFSE-labeled B. subtilis EVs (115\u2009nm \u00b1 27\u2009nm) by Caco-2 cells are time-dependent. To study the transcytosis process we used a transwell system and EVs were quantified in the lower chamber by Fluorescence and Nanoparticle Tracking Analysis measurements. Intact EVs are transported across a polarized cell monolayer at 60-120\u2009min and increased after 240\u2009min with an estimated average uptake efficiency of 30% and this process is dose-dependent. EVs movement into intestinal epithelial cells was mainly through Z axis and scarcely on X and Y axis. This work demonstrates that EVs could be transported across the gastrointestinal epithelium. We speculate this mechanism could be the first step allowing EVs to reach the bloodstream for further delivery up to extraintestinal tissues and organs. The expression and further encapsulation of bioactive molecules into natural nanoparticles produced by probiotic bacteria could have practical implications in food, nutraceuticals and clinical therapies."]

Address: Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina.; Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.; Instituto de Medicina Experimental, Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.; Àrea de Parasitologia, Departament de Farmàcia i TecnologiaFarmacèutica i Parasitologia, Universitat de València, Burjassot, Valencia, Spain.; Joint Research Unit on Endocrinology, Nutrition and Clinical Dietetics, Health Research Institute La Fe, Universitat de Valencia, Valencia, Spain.; Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina. [email protected].; Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina. [email protected].; Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina. [email protected].; Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina. [email protected].

DOI: 10.1038/s41598-020-60077-4

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MeSH Terms: Bacillus subtilis, Caco-2 Cells, Cell Polarity, Cell Proliferation, Cell Survival, Epithelial Cells, Epithelium, Extracellular Vesicles, Functional Food, Humans, Intestines, Microscopy, Confocal, Models, Biological, Probiotics, Transcytosis