June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Central transcorneal freezing alters the proliferation pattern of endothelial cells in the corneal periphery
Author Affiliations & Notes
  • Mette Correll
    Department of Ophthalmology, Rigshospitalet, Copenhagen, Denmark
  • Emmanuel CROUZET
    Corneal Graft Biology, Engineering and Imaging Laboratory, Jean Monnet University, Faculty of Medicine, Saint-Etienne, France
  • Javier Cabrerizo
    Department of Ophthalmology, Rigshospitalet, Copenhagen, Denmark
    Copenhagen Eye Foundation, Copenhagen, Denmark
  • Morten la Cour
    Department of Ophthalmology, Rigshospitalet, Copenhagen, Denmark
  • Philippe GAIN
    Corneal Graft Biology, Engineering and Imaging Laboratory, Jean Monnet University, Faculty of Medicine, Saint-Etienne, France
  • Zhiguo HE
    Corneal Graft Biology, Engineering and Imaging Laboratory, Jean Monnet University, Faculty of Medicine, Saint-Etienne, France
  • Steffen Klarskov Heegaard
    Department of Ophthalmology, Rigshospitalet, Copenhagen, Denmark
    Department of Pathology, Rigshospitalet, Copenhagen, Denmark
  • Jens Folke Kiilgaard
    Department of Ophthalmology, Rigshospitalet, Copenhagen, Denmark
  • Gilles Thuret
    Corneal Graft Biology, Engineering and Imaging Laboratory, Jean Monnet University, Faculty of Medicine, Saint-Etienne, France
  • Footnotes
    Commercial Relationships   Mette Correll, None; Emmanuel CROUZET, None; Javier Cabrerizo, None; Morten la Cour, None; Philippe GAIN, None; Zhiguo HE, None; Steffen Heegaard, None; Jens Kiilgaard, None; Gilles Thuret, None
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 1469. doi:
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      Mette Correll, Emmanuel CROUZET, Javier Cabrerizo, Morten la Cour, Philippe GAIN, Zhiguo HE, Steffen Klarskov Heegaard, Jens Folke Kiilgaard, Gilles Thuret; Central transcorneal freezing alters the proliferation pattern of endothelial cells in the corneal periphery. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1469.

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

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Abstract

Purpose : Studies on corneal endothelial cell (CEC) proliferation related to endothelial injury, can lead to a better understanding of the lack of sufficient regeneration in corneal endothelial dysfunction. We used an in vivo rabbit model of transcorneal freezing to describe CEC proliferation in response to injury and to describe the localization of slow-cycling endothelial cells.

Methods : A central endothelial lesion was induced in 5-weeks old rabbits (n=4) by transcorneal freezing, followed by intraperitoneal injection of 5-Ethynyl-2’-deoxyuridine (EdU). EdU injection was repeated at T24hours and T48 hours to label cells in s-phase during wound healing. Control group underwent the same EdU loading protocol without transcorneal freezing. Animals were euthanized after 2, 5 or 40 days and corneoscleral buttons were flat-mounted. Fluorescence ex vivo confocal microscopy (LSM 710, ZEISS, Oberkochen, Germany) was performed to assess the expression of proliferation marker Ki-67 and EdU-labeling.

Results : Central transcorneal freezing induced denudation of endothelial cells and Descemet's membrane (4 mm Ø). In the central cornea, the CECs adjacent to the denuded area showed extensive proliferation (Ki-67+ and EdU+), covering the lesion within 5 days. No central proliferation (Ki-67- and EdU- ) was detected 40 days after transcorneal freezing in any of the groups. In the periphery, proliferating CECs were present at all time points in both groups, yet during the first 5 days after injury the proliferating CECs were arranged more clustered, compared with the control group, where they continuously showed a scattered pattern. CECs retaining the EdU-label at day 40 were present only in the periphery in both groups.

Conclusions : Proliferation and migration of adjacent CECs seems to be the main mechanism for endothelial repopulation after injury. During central wound healing, alterations in peripheral CEC proliferation towards a clustered cell pattern, indicate central-peripheral communication and possible presence of slow-cycling progenitor cells in the periphery. These data will be usefull for many animal experiments using rabbit, and might explain differences between the rabbit model and human, in which the proliferation of CECs remains much more discreet.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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