Background: It was shown recently that endothelial cell count performed by cornea banks overestimates the real number of endothelial cells. The aim of this study was to investigate the internal quality of preclinical ECD in human donor corneas using two widely used methods for endothelial cell counting, transmitted light microscopy used in organ culture tissue bank and clinically used specular microscopy.
Methods: Twenty human donor corneas that could not be transplanted were included in this analysis. Differences in evaluating endothelial cell density (ECD) and hexagonal endothelial cell ratio (HEX) between clinical specular microscopy (CSM) and corneal bank transmitted light microscope (CBLM) were evaluated as well as differences between automated and manual cell counts.
Results: Automated CBLM showed a higher ECD of 31.85% compared to automated CSM, while manual CBLM counting is 10.51% higher compared to manual CSM (p < 0.01). Further, higher average ECD values result in a higher difference between CSM and CBLM measurements. The manual CBLM ECDs were significantly higher compared to automated derived ECD from CSM (p < 0.01). However, no systematic bias can be detected when comparing the differences of the measurements with the average ECD measurements of both methods.
Conclusion: This preclinical pilot study confirmed a significant higher ECD using transmitted light microscopy in organ culture compared to clinical specular microscopy. This indicates that the early rapid decrease of EC universally observed after surgery might be partly artefactual.
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