Viable endothelial cell density by triple HEC staining of a failed Descemet stripping automated endothelial keratoplasty

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The rate of DSAEK failure decreases during learning curve. It has been previously reported that one of the main cause of rapid DSAEK failure was endothelial cell (EC) loss (Zhang, Arch Ophthalmol 2010). Aim: to report a DSAEK immediate failure and its innovative assessment with determination of the viability of residual EC


In the eye bank, a cornea (donor: 44 years old, organ-culture: 23 days, ECD the day before cut: 2963 cells/mm2, deswelling 21h) was precut with a single pass of a 350μm head (Moria microkeratome). Lamella thickness was 199μm (ultrasound). The recipient had a Fuchs dystrophy. In the operating room, trephination with a Baron punch was eccentric but insertion was uneventful. The graft failed despite 2 rebubbling. A penetrating keratoplasty was performed 2 months later. During surgery the explanted lamella was immediately incubated with Hoechst 33342-Ethidium-Calcein-AM (HEC triple staining, Pipparelli, IOVS 2011) to determine the viability of residual EC. The thickness map of the lamella was then performed with an OCT (Casia, Tomey). The posterior surface of the recipient corneal button was observed with a macroscope


The eccentric trephination caused a important step on 90° of circumference. Unplanned retention of Descemet membrane with guttae was found on the posterior face of the recipient cornea. The viable ECD of the explanted lamella was 585 cells/mm2. Keratocytes of both the lamella and the recipient cornea were viable


Allogenic EC and keratocytes survive on a detached lamella at least during 2 months in the anterior chamber of the recipient. Grant: AOL 2012

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