Proteomics indicated a metabolic shift from oxidative phosphorylation to glycolysis and hypoxia after RF-D/UVA treatment. In contrast, WST-D/NIR stiffening maintained normal respiration and involved extracellular matrix remodeling.

Rabbit corneas were mechanically de-epithelialized, then left untreated (N = 3) or treated with either RF-D/UVA (N = 3) or WST-D/NIR (N = 3). After one week, quantitative proteomics was performed on untreated, RF-D/UVA- and WST-D/NIR-treated corneas. Pathway enrichment analysis was performed to identify the biological processes associated with the treatments. To identify the abundance and spatial distribution of lipids in the untreated, WST-D/NIR- and RF-D/UVA-treated corneal stroma, lipid mass spectrometry imaging was performed together with hematoxylin and eosin staining.

This study aims to elucidate on changes in biological pathways in rabbit corneas induced by two

Between RF-D/UVA- and WST-D/NIR-treated corneas, 37 and 39 proteins, respectively, were differentially expressed compared to untreated corneas (P < 0.05). Pathway enrichment analysis showed the effect of RF-D/UVA treatment on cell metabolism and terminal differentiation of keratocytes, while WST-D/NIR modified extracellular matrix regulation and the mitogen-activated protein kinase signaling cascade. When comparing the RF-D/UVA and WST-D/NIR treatment, 74 proteins were differentially expressed, affecting cellular metabolism and respiration, complement activation, the activation of matrix metalloproteinases, and lipoprotein metabolism. The lipid profile for the RF-D/UVA- and WST-D/NIR-treated stromas were similar, whereas differences were observed comparing both treatments to untreated corneal stroma. Conclusions: Proteomics indicated a metabolic shift from oxidative phosphorylation to glycolysis and hypoxia after RF-D/UVA treatment. In contrast, WST-D/NIR stiffening maintained normal respiration and involved extracellular matrix remodeling.