# Potential Roles of Extracellular Vesicles in Murine Tear Fluids in the Physiology of Corneal Epithelial Cells In Vitro

## TL;DR

This study explores how extracellular vesicles in mouse tear fluids may support the health of corneal cells in a lab setting.

## Contribution

The study identifies exosome-like extracellular vesicles in mouse tear fluids and demonstrates their role in promoting corneal epithelial cell proliferation.

## Key findings

- Mouse tear fluids contain exosome-like extracellular vesicles marked by CD9 and TSG101.
- Tear extracellular vesicles are taken up by corneal epithelial cells and alter gene expression related to cell cycle and membranes.
- Treatment with tear extracellular vesicles increases proliferation of cultured corneal epithelial cells.

## Abstract

Biological extracellular vesicles in tear fluids, such as exosomes, are thought to have physiological functions in the management of healthy ocular surface epithelium, including corneal epithelium. However, the physiological roles of tear extracellular vesicles in the ocular surface remain unclear. In this study, we investigated the physiological function of tear extracellular vesicles in mouse tear fluids in the ocular surface epithelium in vitro. Morphological analysis of the isolated extracellular vesicles from mouse tear fluids was performed using nanoparticle tracking analysis and transmission electron microscopy. The identified particles were characterised by immunoblotting for exosomal markers. After confirming the uptake of tear exosomes in cultured corneal epithelial cells, gene expression changes in mouse cultured corneal epithelial cells after tear exosome treatment were analysed. Immunostaining analysis was performed to confirm cell proliferation in the cultured corneal epithelial cells with tear exosome treatment. Tear fluids from mice contain nanoparticles with exosome-like morphologies, which express the representative exosomal markers CD9 and TSG101. The extracellular vesicles can be taken up by cultivated murine corneal epithelial cells in vitro and induce expression changes in genes related to the cell cycle, cell membranes, microtubules, and signal peptides. Treatment with the tear extracellular vesicles promoted cell proliferation of cultured murine corneal epithelial cells. Our study provides evidence that murine tear fluids contain extracellular vehicles like exosomes and they may contribute to the maintenance of the physiological homeostatic environment of the ocular surface.