Dataset Information

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

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?nm ± 27?nm) 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?min and increased after 240?min 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.

SUBMITTER: Rubio APD

PROVIDER: S-EPMC7033168 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

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

Rubio Ana Paula Domínguez APD Martínez Jimena J Palavecino Marcos M Fuentes Federico F López Christian Miquel Sánchez CMS Marcilla Antonio A Pérez Oscar Edgardo OE Piuri Mariana M

Scientific reports 20200220 1

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 nm ± 27 nm) by Caco-2 cells are time-dependent. To study the transcytosis process we used a transwell system and EVs were q ...[more]

PMID: 32080346

Similar Datasets

Project description:Despite the demonstrated effectiveness of nano-materials for drug delivery to the brain, a comprehensive understanding of their transport processes across the blood brain barrier (BBB) remains undefined. This multidisciplinary study aimed to gain an insight into the transport processes across BBB, focusing on the transcytosis of liposomes and the impact of liposomal pH-sensitivity. Glutathione-PEGylated pH-sensitive (GSH-PEG-pSL) and non pH-sensitive liposomes (GSH-PEG-L) were fluorescently labelled with rhodamine-DOPE and calcein, both impermeable to biomembranes. Following exposure to brain microvascular endothelial cells (hBMECs), the key functional component of the BBB, intracellular trafficking were evaluated by confocal live-cell imaging. The exocytosed liposomes, including naturally-occurring extracellular vesicles (EVs), were collected using differential centrifugation and and characterised regarding the EV yield, morphology and EVs origin using nanoparticle tracking analysis, transmission electron microscopy and flow cytometry. The transcytosis of liposomes through a verified BBB model comprising of hBMECs monolayer was also quantified. GSH-PEG-L was initially retained in the endo-lysosomes before exocytosed while packed in EVs of different sizes (<100 nm to >1 μm) while GSH-PEG-pSL underwent endosome escape with less degree of exocytosis with more fluorescence remaining in the cytoplasm. Compared with the untreated, hBMECs treated with GSH-PEG-L increased the yield of nano-EV and medium-EV by 7.9-fold and 4.6-fold, respectively. Conversely, GSH-pSL-treated cells produced 2.9-fold more nano-EVs but 2-fold less medium-EVs than the control cells. These vesicles were CD144-positive confirming their endothelial cell-origin. GSH-PEG-L demonstrated 2-fold higher efficiencies than GSH-PEG-pSL to cross the in vitro BBB model via exocytosis. Taken together, GSH-PEG-L might utilize EV secretion pathway to achieve transcytosis across brain endothelial cells of the BBB while liposomal pH-sensitivity favors cytoplasmic delivery.

Dataset Information

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

Publications

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

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets