May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
A Single Application of Mitomycin-C Induces DNA Cross-Linking in Corneal Endothelium
Author Affiliations & Notes
  • D. Roh
    UPMC Eye Center, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
  • S. Rhee
    UPMC Eye Center, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
  • A. Cook
    UPMC Eye Center, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
  • A. Joshi
    UPMC Eye Center, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
  • R. Kowolski
    UPMC Eye Center, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
  • D. Dhaliwal
    UPMC Eye Center, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
  • J. Funderburgh
    UPMC Eye Center, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
  • Footnotes
    Commercial Relationships  D. Roh, None; S. Rhee, None; A. Cook, None; A. Joshi, None; R. Kowolski, None; D. Dhaliwal, None; J. Funderburgh, None.
  • Footnotes
    Support  NIH Grants EY09368, EY16415 (JLF), P30-EY08098, Eye Ear Foundation of Pgh, Research To Prevent Blindness
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3959. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      D. Roh, S. Rhee, A. Cook, A. Joshi, R. Kowolski, D. Dhaliwal, J. Funderburgh; A Single Application of Mitomycin-C Induces DNA Cross-Linking in Corneal Endothelium. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3959.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : A single intraoperative application of the DNA cross-linking agent mitomycin-C (MMC) is common for prophylactic haze reduction during corneal refractive surgery. The extent of DNA crosslinking induced in corneal cells by such treatment is unknown. Such crosslinking could affect function of the corneal endothelium (CE) a nondividing layer vital to the transparency of the cornea. This study aims to determine the presence of MMC and MMC-induced DNA crosslinking in CE in both intact globes and in culture.

Methods: : MMC concentration in CE was examined after a 2 min application of a 0.02% solution to intact goat globes. Inhibition of E. coli growth by sliding microtome sections of treated corneas was compared to that of MMC standards. DNA cross-linking was examined in cultured CE treated with MMC at varying times and concentrations and also in intact globes under conditions emulating photorefractive keratectomy: after epithelial debridement, after debridement and laser ablation, and after debridement followed by 24 hour incubation. CE cells were embedded in agarose, spread on slides, lysed, irradiated to induce dsDNA breaks, and electrophoresed under denaturing conditions. DNA from individual cells was stained and photographed. Quantitative image analysis determined cross-linked DNA with reduced electrophoretic mobility, classified as %DNA-in-Head.

Results: : After epithelial exposure to MMC the concentration in the CE determined from a standard curve reached 0.37 microg/mL. In both ocular and cultured CE, all MMC treatments generated more %DNA-in-Head when compared to untreated controls (ANOVA p<0.0001, Dunnett’s p<0.01). In intact globes, increased length of treatment at 0.02% resulted in increased DNA crosslinking. In cultured CE, MMC treatment at concentrations present in the CE also resulted in significant increases in DNA crosslinking.

Conclusions: : A single topical MMC application results in its presence in the corneal endothelium. This exposure leads to significant DNA crosslinking in a time-dependent and concentration-dependent manner in both intact globes and in culture.

Keywords: cornea: endothelium • drug toxicity/drug effects • refractive surgery 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×