# Rat Hair Follicle Stem Cell-Derived Exosomes: Isolation, Characterization and Comparative Analysis of Their In Vitro Wound Healing Potential

## TL;DR

This study explores exosomes from rat hair follicle stem cells for wound healing, showing they could be used as cell-free therapies for skin regeneration.

## Contribution

The first isolation and characterization of exosomes from rat hair follicle stem cells, along with their wound-healing evaluation.

## Key findings

- Exosomes from rat hair follicle stem cells showed pro-regenerative potential in wound healing.
- Secretome and exosomes were found to be cytocompatible and effective in promoting wound closure.
- Exosomal cargo was shown to have therapeutic relevance compared to the parent stem cells.

## Abstract

Stem cell-derived secretome and exosomes present a promising cell-free strategy for tissue repair and wound healing. This study aimed to isolate and characterize, for the first time, exosomes derived from rat hair follicle stem cells (rHFSCs) and to evaluate their wound-healing potential alongside rHFSC secretome. Exosomes were isolated via ultracentrifugation and characterized using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), biomarker profiling and protein quantification. Scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDS) confirmed their spherical morphology, diameter and elemental composition. Protein quantification showed higher protein content in the secretome than in exosomes. RT-PCR and biomarker profiling highlighted the therapeutic relevance of the exosomal cargo compared to parent rHFSCs. Functional analysis of 30 wound-healing biomolecules validated their pro-regenerative potential. Cytocompatibility was confirmed via the PrestoBlue™ viability assay, while scratch assays demonstrated significant wound closure in the treated groups, both with and without mitomycin C. These findings highlight the potential of rHFSC-derived exosomes and secretome as innovative, cell-free therapeutic agents for cutaneous regeneration. This study advances our understanding of their role in wound healing and underscores their broader applicability in regenerative medicine.