A simple <i>ex vivo</i> model for teaching Descemet membrane endothelial keratoplasty

Droutsas, Konstantinos; Petrak, Michael; Melles, Gerrit R.J.; Koutsandrea, Chryssanthi; Georgalas, Ilias; Sekundo, Walter

doi:10.1111/aos.12371

Cited by 14 publications

(14 citation statements)

References 24 publications

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“…The porcine eye model has been used to assess the feasibility of transplanting tissue engineered corneal grafts[ 33 ] and has long been used as a training model for corneal transplantation including DMEK[ 34 ]. The model has good face validity.…”

Section: Discussionmentioning

confidence: 99%

Real-time assessment of corneal endothelial cell damage following graft preparation and donor insertion for DMEK

et al. 2017

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PurposeTo establish a method for assessing graft viability, in-vivo, following corneal transplantation.MethodsOptimization of calcein AM fluorescence and toxicity assessment was performed in cultured human corneal endothelial cells and ex-vivo corneal tissue. Descemet membrane endothelial keratoplasty grafts were incubated with calcein AM and imaged pre and post preparation, and in-situ after insertion and unfolding in a pig eye model. Global, macroscopic images of the entire graft and individual cell resolution could be attained by altering the magnification of a clinical confocal scanning laser microscope. Patterns of cell loss observed in situ were compared to those seen using standard ex-vivo techniques.ResultsCalcein AM showed a positive dose-fluorescence relationship. A dose of 2.67μmol was sufficient to allow clear discrimination between viable and non-viable areas (sensitivity of 96.6% with a specificity of 96.1%) and was not toxic to cultured endothelial cells or ex-vivo corneal tissue. Patterns of cell loss seen in-situ closely matched those seen on ex-vivo assessment with fluorescence viability imaging, trypan blue/alizarin red staining or scanning electron microscopy. Iatrogenic graft damage from preparation and insertion varied between 7–35% and incarceration of the graft tissue within surgical wounds was identified as a significant cause of endothelial damage.ConclusionsIn-situ graft viability assessment using clinical imaging devices provides comparable information to ex-vivo methods. This method shows high sensitivity and specificity, is non-toxic and can be used to evaluate immediate cell viability in new grafting techniques in-vivo.

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Section: Discussionmentioning

confidence: 99%

Real-time assessment of corneal endothelial cell damage following graft preparation and donor insertion for DMEK

et al. 2017

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“…Despite reports emerging from different tertiary centers confirming the short and safe learning curve of DMEK 2,8,10,11,15,23 and also a recently described wetlab model for DMEK training, 24 graft preparation and atraumatic surgery are still considered as technically challenging. 22,25 Thus, other techniques such as "thin" or "ultrathin" DSAEK have been developed, whose efficacy however still needs to be evaluated and compared with DMEK.…”

Section: Discussionmentioning

confidence: 99%

Visual Outcomes After Descemet Membrane Endothelial Keratoplasty Versus Descemet Stripping Automated Endothelial Keratoplasty—Comparison of Specific Matched Pairs

Droutsas

Lazaridis

Papaconstantinou

et al. 2016

Cornea

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“…The lens epithelial cells deposit extracellular matrix onto the capsule, and thus, the LC gradually thickens with age (Danysh & Duncan, ). The capsule is inherently transparent and can form a roll when submersed in water, behaving similar to the Descemet membrane making it an attractive scaffold option (Droutsas et al, ). Anterior capsular material could potentially be obtained as a by‐product of cataract surgery, one of the most frequently performed surgeries in the world.…”

Section: Introductionmentioning

confidence: 99%

Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering

Bogerd

Dhubhghaill

Zakaria

2018

J Tissue Eng Regen Med

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The idea of transplanting a sheet of laboratory‐grown corneal endothelium dates back to 1978; however, the ideal scaffold is still lacking. We hypothesized that human crystalline lens capsules (LCs) could qualify as a scaffold and aimed to characterize the properties of this material for endothelial tissue engineering. LCs were isolated from donor eyes, stored at −80 °C, and decellularized with water and trypsin‐EDTA. The decellularization was investigated by nuclear staining and counting and the capsule thickness was determined by optical coherence tomography and compared with Descemet's membrane (DM). Transparency was examined by spectrometry, and collagenase degradation was performed to evaluate its resistance to degradation. Cell‐scaffold interaction was assessed by measuring focal adhesions surface area on LC and plastic. Finally, primary corneal endothelial cells were grown on LCs to validate the phenotype. Trypsin‐EDTA decellularized most effectively, removing 99% of cells. The mean LC thickness was 35.76 ± 0.43 μm, whereas DM measured 25.93 ± 0.26 μm (p < .0001). Light transmission was 90% for both LC and DM. On a collagenase challenge, LC and amniotic membrane were digested after 13 hr, whereas DM was digested after 17 hr. The surface area of focal adhesions for cells grown on coated LCs was at least double that compared with other conditions, whereas tight junctions, ion pumps, and hexagonal morphology were well maintained when endothelial cells were cultured on LCs. In conclusion, LCs demonstrate excellent scaffolding properties for tissue engineering and sustain the cell phenotype and can be considered a suitable substrate for ocular tissue engineering or as a template for future scaffolds.

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A simple ex vivo model for teaching Descemet membrane endothelial keratoplasty

Cited by 14 publications

References 24 publications

Real-time assessment of corneal endothelial cell damage following graft preparation and donor insertion for DMEK

Real-time assessment of corneal endothelial cell damage following graft preparation and donor insertion for DMEK

Visual Outcomes After Descemet Membrane Endothelial Keratoplasty Versus Descemet Stripping Automated Endothelial Keratoplasty—Comparison of Specific Matched Pairs

Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering

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