Intrastromal keratotomy with femtosecond laser avoids profibrotic TGF-beta1 induction

Invest Ophthalmol Vis Sci. 2009 Aug;50(8):3688-95. doi: 10.1167/iovs.08-2699. Epub 2009 Apr 22.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Corneal Stroma / metabolism*
  • Corneal Stroma / surgery*
  • Endothelium, Corneal / metabolism
  • Epithelium, Corneal / metabolism
  • Fluorescent Antibody Technique, Indirect
  • Gene Expression Regulation / physiology*
  • Lasers, Excimer / therapeutic use*
  • Photorefractive Keratectomy / methods*
  • RNA / isolation & purification
  • RNA, Messenger / metabolism
  • Rabbits
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transforming Growth Factor beta1 / genetics*
  • Transforming Growth Factor beta1 / metabolism
  • Up-Regulation

Substances

  • RNA, Messenger
  • Transforming Growth Factor beta1
  • RNA