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Mor Dickman, Jurriaan Brekelmans, Petra Steijger-Vermaat, Teja Wesseling, Friso W.F. van Marion, Frank Van den Biggelaar, Tos TJM Berendschot, Rudy M Nuijts; Pre-Descemetic Automated Endothelial Keratoplasty (PDAEK), towards an automated DMEK preparation technique. Invest. Ophthalmol. Vis. Sci. 2014;55(13):2699.
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© ARVO (1962-2015); The Authors (2016-present)
One of the main limitations to wide-spread adoption of Descemet Membrane Endothelial Keratoplasty (DMEK) is the lack of a standardized automated technique for graft preparation. In the current study we set out to establish a proof of concept for automated harvesting of planar pre-Descemetic (<50μm) endothelial grafts using a novel vacuum assisted microkeratome system.
Sixty-nine organ cultured human donor corneas unsuitable for transplantation were dissected using a prototype linear microkeratome system (Gebauer SLc Expert; Gebauer; Germany). Following epithelial removal, donor corneas were applanated using vacuum (600 mbar) against a porous rigid-reference-member (RRM) designed to correct for central to peripheral thickness differences in the donor cornea (figure 1). Thickness measurements were obtained by Fourier domain Anterior-Segment Optical-Coherence-Tomography (Casia SS-1000; Tomey; Japan). Endothelial cell density was determined by Trypan blue exclusion, followed by triple Hoechst-Ethidium-Calcein (HEC) labeling for pan-corneal endothelial viability assessment.
Mean post-cut endothelial graft thickness measured 40 (±7) μm, significantly thinner than 50μm (p<0.05). RRM application resulted in significantly planar graft thickness profiles compared with the meniscus shape observed without RRM application (p<0.05) (figure 2). Mean post-cut endothelial cell density measured 2245 (±175) cells/mm2 with an average 70% (±10%) pan-corneal endothelial viability.
This study established a proof of concept for reproducible automated vacuum assisted harvesting of planar pre-Descemetic endothelial grafts with satisfactory endothelial cell density and viability, addressing many of the limitations of current mechanical microkeratomes and introducing a novel technique for endothelial graft preperation (PDAEK).
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