July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Laminins, and early corneal epithelial basement membrane (EBM) regeneration
Author Affiliations & Notes
  • Rodrigo Carlos De Oliveira
    Cole eye institute, Cleveland Clinic Foundation, Cleveland, Ohio, United States
  • Paramananda Saikia
    Cole eye institute, Cleveland Clinic Foundation, Cleveland, Ohio, United States
  • Steven E Wilson
    Cole eye institute, Cleveland Clinic Foundation, Cleveland, Ohio, United States
  • Footnotes
    Commercial Relationships   Rodrigo Carlos De Oliveira, None; Paramananda Saikia, None; Steven Wilson, None
  • Footnotes
    Support  Supported by NEI RO1 EY10056 (SEW) and Research to Prevent Blindness, New York, NY.
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4663. doi:
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    • Get Citation

      Rodrigo Carlos De Oliveira, Paramananda Saikia, Steven E Wilson; Laminins, and early corneal epithelial basement membrane (EBM) regeneration. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4663.

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

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Abstract

Purpose : To evaluate the expression of laminin alpha 5 (LAM alpha 5) and laminin beta 3 (LAM beta 3) during EBM regeneration.

Methods : NZW rabbits had -4.5 photorefractive keratectomy (PRK) or -9.0 PRK in one eye with a VISX S4 IR excimer laser. Rabbits were sacrificed at 12 hrs, 24 hrs, and 48 hrs after surgery, with 4 eyes at each time point in each group. One rabbit cornea had 9mm epithelial scrape only (12 hrs). Corneas were cryofixed in OCT and had immunohistochemistry for LAM alpha 5 (antibody a gift from Peter Yurchenco) and LAM beta 3.

Results : Control corneas had a continuous monolayers of LAM alpha 5 and LAM beta 3 at the level of the EBM. At 12- and 24-hrs after PRK, before the epithelium had closed, we noted in all corneas, a monolayer of LAM alpha 5 (green) was present on the surface of the bare stroma (Fig. 1). In some corneas at 12- and 24-hrs, both a monolayer of LAM alpha 5 and LAM beta 3 were present on the surface of the bare stroma (Fig. 2). In all 48-hr PRK corneas, the epithelium had closed, and there were layers of LAM alpha 5 and LAM beta 3 at the level of EBM, and high levels of LAM beta 3 in basal epithelial cells. There was a continuous layer of LAM alpha 5, but not LAM beta 3, on the bare stroma of the cornea that had epithelial scrape only. No stromal cell production of LAM alpha 5 or LAM beta 3 was detected.

Conclusions : LAM alpha 5 (likely a component of self-polymerizing laminins 511 and/or 521) appeared on the surface of the bare stroma of all corneas after -4.5D or -9D PRK or epithelial scrape. Since no stromal cells were seen to produce LAM alpha 5, and no monolayer of epithelium was present, this LAM alpha 5 may have been deposited from tears. Some corneas at 12- and 24-hrs after PRK, also had LAM beta 3 (likely a component of laminin 332 found at high levels in corneas) that joined laminins 511 and/or 521 that began regeneration of the nascent EBM. LAM beta 3 also appears to transfer to the bare stromal surface via tears since no LAM beta 3 was detected in stromal cells and regenerating epithelial cells producing LAM beta 3 remained peripheral at these time points. By 48-hours after PRK, continuous layers of LAM alpha 5 and LAM beta 3 were present in EBM, with LAM beta 3 also present in basal epithelial cells. There was no difference at these early time points between -4.5D and -9D PRK.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

IHC for LAM alpha 5 and LAM alpha 3 in corneas after PRK.

IHC for LAM alpha 5 and LAM alpha 3 in corneas after PRK.

 

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