April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Collagen Cross-Linking Procedure Induces Severe Damage in Corneas Without Bowman’s Layer, in Hens
R. Cantalapiedra; T. Blanco; J. Hincapie; P. Perez-Merino; S. Mar; L. Ibares-Frias; P. Gallego; S. del Olmo-Aguado; I. Alcalde; J. Merayo-Lloves
Author Affiliations & Notes
  • R. Cantalapiedra
    IOBA,
    University of Valladolid, Valladolid, Spain
  • T. Blanco
    Schepens Eye Research Institute, Harvard University, Boston, Massachusetts
  • J. Hincapie
    IOBA,
    University of Valladolid, Valladolid, Spain
  • P. Perez-Merino
    IOBA,
    Cell Biology and Histology,
    University of Valladolid, Valladolid, Spain
  • S. Mar
    IOBA,
    University of Valladolid, Valladolid, Spain
  • L. Ibares-Frias
    IOBA,
    University of Valladolid, Valladolid, Spain
  • P. Gallego
    IOBA,
    Cell Biology and Histology,
    University of Valladolid, Valladolid, Spain
  • S. del Olmo-Aguado
    IOBA,
    Cell Biology and Histology,
    University of Valladolid, Valladolid, Spain
  • I. Alcalde
    IOBA,
    Cell Biology and Histology,
    University of Valladolid, Valladolid, Spain
  • J. Merayo-Lloves
    IOBA,
    University of Valladolid, Valladolid, Spain
  • Footnotes
    Commercial Relationships  R. Cantalapiedra, None; T. Blanco, None; J. Hincapie, None; P. Perez-Merino, None; S. Mar, None; L. Ibares-Frias, None; P. Gallego, None; S. del Olmo-Aguado, None; I. Alcalde, None; J. Merayo-Lloves, None.
  • Footnotes
    Support  Supported by a PROFIT Grant CIT-300100-2007-50 Ministry of Health of Spain
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 914. doi:
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      R. Cantalapiedra, T. Blanco, J. Hincapie, P. Perez-Merino, S. Mar, L. Ibares-Frias, P. Gallego, S. del Olmo-Aguado, I. Alcalde, J. Merayo-Lloves; Collagen Cross-Linking Procedure Induces Severe Damage in Corneas Without Bowman’s Layer, in Hens. Invest. Ophthalmol. Vis. Sci. 2009;50(13):914.

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Abstract

Methods: : In a 6,00 mm diameter area, the Bowman layer of both eyes of 28 hens was removed with an excimer laser. After two months the right corneas, were UVA-irradiated (360-380 nm, 3 mW/cm2), in a 6,00 mm diameter deepithelized area, during 30 minutes. A 0,1% riboflavin-20% dextran solution was administered 5 minutes before irradiation and then every 5 minutes during the UVA irradiation. The left fellow control corneas were only debridement. Then, animals were allowed to heal for up to 1 month and euthanized in different after 6, 12 and 24 hours, 3, 5, 15 and 30 days, being the corneas fixed for pathology. Cell death was evaluated with TUNEL assay. Anti BrdU for cell proliferation and expression of -SMA indicative of myofibroblast differentiation were evaluated by indirect immunofluorescence microscopy.

Results: : Differences in the time of epithelial closure (48±8 hours) were not observed although a great grade of edema was observed in the CXL corneas (p<0.05). The corneal thickness remains higher in the right corneas (p<0.05). Endothelial damage was observed in 100% of CXL corneas and in none of the control. Although subepithelial haze was not obserbed in any cornea, endothelial haze was observed in 100% of right corneas. Presence of inflammatory response was observed higher in the anterior chamber of irradiated eyes (p<0,05).Keratocytes and endothelial cells were full disappeared in the irradiated area in the CXL corneas and only in the anterior stromal third in the controls. Proliferation of new endothelial cells and keratocytes was observed at the third day after treatment. At the fifth day presence of myofibroblast was observed closing the endothelial holds. At the fifteenth day the endothelium was full restored again with new endothelial cells. At day thirtieth all the corneas appears with a normal cell density and subepithelial fibrosis was not seen.

Keywords: cornea: stroma and keratocytes • cornea: endothelium • radiation damage: light/UV 
© 2009, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
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