May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Regulation of corneal epithelial homeostasis in transplantable corneas
Author Affiliations & Notes
  • J. Soltaninia
    Optometry, Cardiff University, Cardiff, United Kingdom
  • M. Boulton
    Optometry, Cardiff University, Cardiff, United Kingdom
  • W.J. Armitage
    Ophthalmology, Bristol University, Bristol, United Kingdom
  • J. Albon
    Optometry, Cardiff University, Cardiff, United Kingdom
  • Cell and Molecular Biology Unit
    Optometry, Cardiff University, Cardiff, United Kingdom
  • Footnotes
    Commercial Relationships  J. Soltaninia, None; M. Boulton, None; W.J. Armitage, None; J. Albon, None.
  • Footnotes
    Support  NERC
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 1460. doi:
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      J. Soltaninia, M. Boulton, W.J. Armitage, J. Albon, Cell and Molecular Biology Unit; Regulation of corneal epithelial homeostasis in transplantable corneas . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1460.

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

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Abstract

Abstract: : Purpose:To obtain a better understanding of corneal epithelial cell homeostasis during organ culture storage. Methods:Porcine corneas were maintained in immersion organ culture for varying periods of time. For proliferation studies BrdU at 1/500 dilution was added. Corneas were then stored in an incubator at 34° C for up to 14 days. At each time point the cornea was bisected: one half was fixed for wax sectioning and the other half snap frozen for cryosectioning. General morphology, epithelial stratification and cell density were assessed by light and fluorescence microscopy using Leica Qfluoro image analysis software. Keratocyte number and density was quantified in the anterior, posterior and middle regions of the corneal stroma. The proliferative status of the epithelium was assessed as a function of BrdU incorporation and quantified in central, peripheral and limbal regions. Results:A significant decrease in corneal epithelial thickness was demonstrated with increasing time of organ culture from a maximum value of 45µm ± 3.6 at 0 day to a minimum of 7.7µm ± 0.6 at 14 days (P<0.0001). Stromal thickness increased from 691µm ± 53 at 0 day to a maximum of 1569µm ± 126 at 14 days organ culture storage (P<0.0001). There was also an apparent decrease in keratocyte density in anterior, posterior and middle central stromal (P<0.0001). The BrdU labelling index as a measure of the proliferation rate was not equally distributed across the cornea. Proliferation was greatest in the peripheral epithelium at 12 days (27.7% ± 8.6%) and the labelling index appeared to increase in the peripheral (r=0.79), central (r=0.62) and limbal (r=0.67) region with organ culture storage time (P<0.05). Conclusions:Porcine cornea is a good model for organ culture storage. Furthermore like human corneas changes occur in epithelial cell homeostasis and stromal keratocytes with time in culture. This may result in barrier dysfunction and therefore indirectly endothelial cell death that is responsible for the 30% discard rate of corneas stored for transplantation.

Keywords: cornea: epithelium • cornea: stroma and keratocytes • transplantation 
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