Cultured media from adipose derived stem cells promote corneal epithelial wound healing via increased cell proliferation and DNA metabolic process

Abstract

Purpose : This study aimed to determine the molecular mechanisms involved with using cultured media from adipose derived stem cells (CM-ADSCs) for corneal epithelial cell wound healing compared to platelet-rich plasma (PRP).

Methods : The efficacies of CM-ADSCs, PRP, and phosphate-buffered saline (PBS) were compared using an in vitro human corneal epithelial wound healing assay. The in vitro human corneal epithelial wounds were 70-µm wide for each well (96-well plate) containing cultured human corneal epithelial cells (HCECs). Following wounding, culture media were replaced with PBS, PRP, and CM-ADSCs, and diluted in corneal epithelial cell basal medium. Wound images were taken every 3h, and the area of the wound was calculated using the ImageJ software. RNA-sequencing analysis was performed on the collected HCECs after 12 hours post wound preparation.

Results : In the in vitro human corneal epithelial wound healing assay, the CM-ADSCs group significantly promoted wound healing compared to the PBS group and the PRP group (N = 6, P < 0.001, P = 0.020, respectively). The CM-ADSCs group (21.0±3.3 hours) had a significantly faster wound closure time compared to the PBS group (55.0±3.1 hours, P<0.001) and the PRP group (26.5±3.5 hours, P<0.001). Differentially expressed genes analysis showed that the expressions increased 1503 genes and decreased 1417 genes were identified in the CM-ADSCs group compared to PRP group. The Gene Ontology term enrichment analysis results revealed significantly enriched biological processes in increased DNA metabolic process and positive regulation of cell cycle process in CM-ADSCs group compared to the PRP group.

Conclusions : CM-ADSCs may promote wound healing of HCECs via increased cell proliferation and DNA metabolic process.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.