Adipose-derived mesenchymal stem cell-conditioned medium promotes corneal epithelial cell wound healing in vitro through cell-cycle activation.

PURPOSE: Adipose-derived mesenchymal stem cell-conditioned medium (ADSC-CM) shows potential for promoting corneal epithelial wound healing; however, its effects on human corneal epithelium and underlying mechanisms remain unclear. This study aimed to evaluate the efficacy and mechanism of action of ADSC-CM in corneal epithelial wound healing.

METHODS

Growth factor levels (epidermal growth factor, transforming growth factor beta 1 [TGF-β1], and fibronectin) in ADSC-CM and platelet-rich plasma (PRP) were measured using enzyme-linked immunosorbent assay. The effects of 10% ADSC-CM, PRP, and phosphate-buffered saline on corneal epithelial wound healing were assessed in human corneal epithelial cells, whereas angiogenic effects were evaluated in human umbilical vein endothelial cells. The mRNA expression levels of proliferation, angiogenesis, lymphangiogenesis, and inflammation markers were analyzed using RT-qPCR. RNA sequencing and subsequent Gene Ontology (GO) enrichment analysis and Gene Set Enrichment Analysis (GSEA) were performed to identify differentially expressed genes and enriched pathways.

RESULTS

ADSC-CM showed higher epidermal growth factor but lower TGF-β1 and fibronectin levels than PRP and significantly promoted corneal epithelial wound healing compared with PRP and phosphate-buffered saline. ADSC-CM also suppressed human umbilical vein endothelial cell migration. RT-qPCR showed increased expression of EGF, IGF2, and FGF1 and decreased expression of IL1B and TNFA in ADSC-CM. Differential gene expression analysis revealed 3849 altered genes; GO and GSEA indicated upregulation of cell-cycle process-related genes and downregulation of inflammation- and angiogenesis-related genes.

CONCLUSIONS

ADSC-CM enhanced corneal epithelial wound healing by promoting proliferation and suppressing inflammation and angiogenesis, highlighting its therapeutic potential in refractory kerato-conjunctival epithelial disorders.