PURPOSE: To examine expression of the profibrotic cytokine TGF-beta1 after selective intrastromal corneal injury with the use of a femtosecond laser. METHODS: Rabbits underwent monocular intrastromal keratotomy at a preoperatively determined corneal depth of 160 to 200 mum with the use of a femtosecond laser. Femtosecond laser-induced TGF-beta1 expression was compared in nonoperated control eyes and eyes treated with photorefractive keratectomy (PRK). Follow-up examinations were performed 1, 3, 7, and 28 days after surgery. TGF-beta1 protein was identified by immunofluorescence labeling. With the use of laser-capture microdissection, epithelial, stromal, and endothelial cell layers were collected, and changes in TGF-beta1 mRNA expression were quantified with quantitative RT-PCR. RESULTS: TGF-beta1 mRNA and protein expression did not significantly increase after intrastromal femtosecond laser keratotomy. In contrast, TGF-beta1 was induced in corneal epithelial and stromal cells after PRK and showed up to 23-fold higher TGF-beta1 mRNA levels compared with control corneas. The increase of TGF-beta1 mRNA levels after PRK was accompanied by increased TGF-beta1 protein production. CONCLUSIONS: Isolated stromal injury with a femtosecond laser does not result in induction of the profibrotic cytokine TGF-beta1. Because TGF-beta1 has been implicated in a fibrotic response of the corneal stroma to injury, absence of TGF-beta1 induction argues for a favorable wound-healing response. These findings support highly selective intrastromal procedures in refractive surgery.
PURPOSE: To examine expression of the profibrotic cytokine TGF-beta1 after selective intrastromal corneal injury with the use of a femtosecond laser. METHODS:Rabbits underwent monocular intrastromal keratotomy at a preoperatively determined corneal depth of 160 to 200 mum with the use of a femtosecond laser. Femtosecond laser-induced TGF-beta1 expression was compared in nonoperated control eyes and eyes treated with photorefractive keratectomy (PRK). Follow-up examinations were performed 1, 3, 7, and 28 days after surgery. TGF-beta1 protein was identified by immunofluorescence labeling. With the use of laser-capture microdissection, epithelial, stromal, and endothelial cell layers were collected, and changes in TGF-beta1 mRNA expression were quantified with quantitative RT-PCR. RESULTS: TGF-beta1 mRNA and protein expression did not significantly increase after intrastromal femtosecond laser keratotomy. In contrast, TGF-beta1 was induced in corneal epithelial and stromal cells after PRK and showed up to 23-fold higher TGF-beta1 mRNA levels compared with control corneas. The increase of TGF-beta1 mRNA levels after PRK was accompanied by increased TGF-beta1 protein production. CONCLUSIONS: Isolated stromal injury with a femtosecond laser does not result in induction of the profibrotic cytokine TGF-beta1. Because TGF-beta1 has been implicated in a fibrotic response of the corneal stroma to injury, absence of TGF-beta1 induction argues for a favorable wound-healing response. These findings support highly selective intrastromal procedures in refractive surgery.