Effects and molecular mechanisms of cultured media from adipose-derived stem cells during corneal wound healing
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Purpose
This study aimed to investigate the effects and molecular mechanisms of cultured media from adipose-derived stem cells (CM-ADSCs) for corneal wound healing compared to platelet-rich plasma (PRP).
Methods
The concentrations of growth factor of CM-ADSCs and PRP were measured by ELISA. The efficacies of CM-ADSCs, PRP, and PBS were compared using an in vitro wound healing assay with human corneal epithelial cells (HCECs). RT-qPCR and RNA-sequencing were performed on HECEs from in vitro wound healing assay. RT-qPCR was used to evaluate mRNA expression of factors related to corneal epithelial proliferation, differentiation, and inflammation. RNA sequencing was performed to identify differentially expressed genes (DEGs) and Gene Ontology (GO) and gene set enrichment analysis (GSEA).
Results
EGF levels were higher with CM-ADSCs compared to PRP. CM-ADSCs promoted corneal epithelial wound healing compared to other groups. RT-qPCR showed higher expression of Egf, Igf2, and Fgf2 and lower Il1b and Tnfa with CM-ADSCs compared to PRP. DEG analysis revealed altered expression of 3,849 genes with CM-ADSCs compared to PRP. GO analysis and GSEA revealed that CM-ADSCs, compared to PRP, exhibited enriched gene expression associated with biological processes such as cell adhesion, cell division, and positive regulation of gene expression, while demonstrating reduced activity related to inflammation and angiogenesis.
Conclusion
CM-ADSCs may promote corneal epithelial wound healing by enhancing cell proliferation and DNA metabolic processes, while also reducing inflammation and angiogenic effects compared to PRP. This suggests a unique advantage in minimizing inflammation during corneal epithelial wound healing.