title: Quantitative assessment of ultrastructure and light scatter in mouse corneal debridement wounds.
abstract: OBJECTIVE
  The mouse has become an important wound healing model with which to study corneal fibrosis, a frequent complication of refractive surgery. The aim of the current study was to quantify changes in stromal ultrastructure and light scatter that characterize fibrosis in mouse corneal debridement wounds.
  
  
  METHODS
  Epithelial debridement wounds, with and without removal of basement membrane, were produced in C57BL/6 mice. Corneal opacity was measured using optical coherence tomography, and collagen diameter and matrix order were quantified by x-ray scattering. Electron microscopy was used to visualize proteoglycans. Quantitative PCR (Q-PCR) measured mRNA transcript levels for several quiescent and fibrotic markers.
  
  
  RESULTS
  Epithelial debridement without basement membrane disruption produced a significant increase in matrix disorder at 8 weeks, but minimal corneal opacity. In contrast, basement membrane penetration led to increases in light scatter, matrix disorder, and collagen diameter, accompanied by the appearance of abnormally large proteoglycans in the subepithelial stroma. This group also demonstrated upregulation of several quiescent and fibrotic markers 2 to 4 weeks after wounding.
  
  
  CONCLUSIONS
  Fibrotic corneal wound healing in mice involves extensive changes to collagen and proteoglycan ultrastructure, consistent with deposition of opaque scar tissue. Epithelial basement membrane penetration is a deciding factor determining the degree of ultrastructural changes and resulting opacity.
authors.affiliation: Structural Biophysics Group, School of Optometry and Vision Sciences, Cardiff University, Cardiff, United Kingdom. bootec@cf.ac.uk
authors.forename: Craig
authors.initials: C
authors.lastname: Boote
authors.forename: Yiqin
authors.initials: Y
authors.lastname: Du
authors.forename: Sian
authors.initials: S
authors.lastname: Morgan
authors.forename: Jonathan
authors.initials: J
authors.lastname: Harris
authors.forename: Christina S
authors.initials: CS
authors.lastname: Kamma-Lorger
authors.forename: Sally
authors.initials: S
authors.lastname: Hayes
authors.forename: Kira L
authors.initials: KL
authors.lastname: Lathrop
authors.forename: Danny S
authors.initials: DS
authors.lastname: Roh
authors.forename: Michael K
authors.initials: MK
authors.lastname: Burrow
authors.forename: Jennifer
authors.initials: J
authors.lastname: Hiller
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Terrill
authors.forename: James L
authors.initials: JL
authors.lastname: Funderburgh
authors.forename: Keith M
authors.initials: KM
authors.lastname: Meek
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000473:pathology / Q000601:surgery / Q000648:ultrastructure; D015415:Biomarkers / Q000032:analysis; D003094:Collagen / Q000648:ultrastructure*; D003315:Cornea / Q000473:pathology / Q000601:surgery* / Q000648:ultrastructure; D065306:Corneal Injuries; D003318:Corneal Opacity / Q000473:pathology; D003319:Corneal Stroma / Q000503:physiopathology / Q000648:ultrastructure; D003646:Debridement / Q000009:adverse effects*; D019573:Epithelium, Corneal / Q000293:injuries / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D005355:Fibrosis; D008027:Light; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis / Q000096:biosynthesis; D000077278:RNA, Mitochondrial; D012542:Scattering, Radiation; D041623:Tomography, Optical Coherence; D014945:Wound Healing / Q000502:physiology
chemical_list: D015415:Biomarkers; D000077278:RNA, Mitochondrial; D012313:RNA; D003094:Collagen