Short-Term Effect of Rose Bengal Photodynamic Therapy (RB-PDT) on Collagen I, Collagen V, NF-κB, LOX, TGF-β and IL-6 Expression of Human Corneal Fibroblasts, In Vitro .

PURPOSE: To investigate collagen I, collagen V, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), lysyl oxidase (LOX), transforming growth factor β1 (TGF-β1) and interleukin-6 (IL-6) expression in healthy and keratoconus human corneal fibroblasts (HCFs and KC-HCFs), 24 h after Rose Bengal photodynamic therapy (RB-PDT).

METHODS: HCFs were isolated from healthy human corneal donors ( n  = 5) and KC-HCFs from elective penetrating keratoplasties ( n  = 5). Both cell cultures underwent RB-PDT (0.001% RB concentration, 0.17 J/cm2 fluence) and 24 h later collagen I, collagen V, NF-κB, LOX, TGF-β1 and IL-6 mRNA and protein expression have been determined using qPCR and Western blot, IL-6 concentration in the cell culture supernatant by ELISA.

RESULTS: TGF-β1 mRNA expression was significantly lower ( p  = 0.02) and IL-6 mRNA expression was significantly higher in RB-PDT treated HCFs ( p  = 0.01), than in HCF controls. COL1A1, COL5A1 and TGF-β1 mRNA expression was significantly lower ( p  = 0.04; p  = 0.02 and p  = 0.003) and IL-6 mRNA expression was significantly higher ( p  = 0.02) in treated KC-HCFs, than in KC-HCF controls. TGF-β1 protein expression in treated HCFs was significantly higher than in HCF controls ( p  = 0.04). IL-6 protein concentration in the HCF and KC-HCF culture supernatant after RB-PDT was significantly higher than in controls ( p  = 0.02; p  = 0.01). No other analyzed mRNA and protein expression differed significantly between the RB-PDT treated and untreated groups.

CONCLUSIONS: Our study demonstrates that RB-PDT reduces collagen I, collagen V and TGF-β1 mRNA expression, while increasing IL-6 mRNA and protein expression in KC-HCFs. In HCFs, RB-PDT increases TGF-β1 and IL-6 protein level after 24 h.