Desiccating Stress-Induced MMP Production and Activity Worsens Wound Healing in Alkali-Burned Corneas

Invest Ophthalmol Vis Sci. 2015 Jul;56(8):4908-18. doi: 10.1167/iovs.15-16631.

Abstract

Purpose: To evaluate the effects of dry eye on ocular surface protease activity and sight threatening corneal complications following ocular surface chemical injury.

Methods: C57BL/6 mice were subjected to unilateral alkali burn (AB) with or without concomitant dry eye for 2 or 5 days. Mice were observed daily for appearance of corneal perforation. Whole corneas were harvested and lysed for RNA extraction. Quantitative real-time PCR was performed to measure expression of inflammation cytokines, matrix metalloproteinases (MMP). Matrix metalloproteinase-9 activity, gelatinase activity, and myeloperoxidase (MPO) activity were evaluated in corneal lysates. Presence of infiltrating neutrophils was evaluated by immunohistochemistry and flow cytometry.

Results: Eyes subjected to the combined model of AB and dry eye (CM) had 20% sterile corneal perforation rate as soon as 1 day after the initial injury, which increased to 35% by 5 days, delayed wound closure and increased corneal opacity. Increased levels of IL-1β, -6, and MMPs-1, -3, -8, -9, and -13, and chemokine (C-X-C motif) ligand 1 (CSCL1) transcripts were found after 2 days in CM compared with AB corneas. Increased MMP-1, -3, -9, and -13 immunoreactivity and gelatinolytic activity were seen in CM corneas compared with AB. Increased neutrophil infiltration and MPO activity was noted in the CM group compared with AB 2 days post injury.

Conclusions: Desiccating stress worsens outcome of ocular AB, creating a cytokine and protease storm with greater neutrophil infiltration, increasing the risk of corneal perforation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Alkalies / toxicity
  • Animals
  • Burns, Chemical / enzymology
  • Burns, Chemical / genetics*
  • Burns, Chemical / pathology
  • Disease Models, Animal
  • Disease Progression
  • Eye Burns / chemically induced
  • Eye Burns / enzymology
  • Eye Burns / genetics*
  • Eye Burns / pathology
  • Flow Cytometry
  • Gene Expression Regulation*
  • Matrix Metalloproteinase 9 / biosynthesis
  • Matrix Metalloproteinase 9 / genetics*
  • Mice
  • Mice, Inbred C57BL
  • Oxidative Stress*
  • RNA / genetics*
  • Real-Time Polymerase Chain Reaction
  • Wound Healing*

Substances

  • Alkalies
  • RNA
  • Matrix Metalloproteinase 9
  • Mmp9 protein, mouse