Pan-Corneal Endothelial Viability Assessment: Application to Endothelial Grafts Predissected by Eye Banks

Abstract

Standard counting by eye banks overestimates the actual pool of viable endothelial cells. This may be the main explanation for the initially rapid decrease in ECD universally described in patients after all types of keratoplasty. Early low postoperative ECD may indicate that surgeons graft fewer living cells than the eye banks' ECD let suppose, rather than a massive pre- and postoperative cell death. The novel concept of viable ECD can be useful for assessing all types of corneal processing.

Significant differences between specular microscopy and corneal bank endothelial cell counts – a pilot study

“…This is confirmed by Pipparelli et al. () who showed a higher ECD during organ culture, suggesting that the number of ECs at the time of transplantation is less than surgeons think. Even so, Szaflik et al.…”

Significant differences between specular microscopy and corneal bank endothelial cell counts – a pilot study

“…This is confirmed by Pipparelli et al. () who showed a higher ECD during organ culture, suggesting that the number of ECs at the time of transplantation is less than surgeons think. Even so, Szaflik et al.…”

Very Early Endothelial Cell Loss After Simultaneous Corneal Autograft and Allograft

“…This very early cell loss is the major factor in the rapid component of the cell loss biphasic curve after a PKP. 7 For PKP with OC corneas, we found a difference of 12% 6 9% (mean 6 SD) between the ebECD and vECD. This ECD decrease reached the cell loss usually admitted at 6 months (228.8% for Bertelmann et al 20 ) and 1 year in normal risk PKP after OC (239.4% for Harper et al, 21 or 229.7% for Borderie et al 3 ).…”

Vulnerability of corneal endothelial cells to mechanical trauma from indentation forces assessed using contact mechanics and fluorescence microscopy

“…However, differing from previous work, we calculated injured CEC densities by acquiring high-resolution fluorescence micrographs (1990×1440 pixels, 905.75 nm/pixel scale) of calcein/PI-stained buttons, counting the number of PI-labelled CEC nuclei and dividing by the contact area. This method contrasts the more common method of staining buttons with trypan blue/alizarin red S and determining the stained area fraction after thresholding (Pipparelli et al, 2011; Saad et al, 2008). This distinct approach was taken for two reasons: first, our main focus was on assessing damage at sites of indentation only; second, the high resolution of our images allowed for reliable visualization and quantification of individual CEC nuclei.…”

Significant differences between specular microscopy and corneal bank endothelial cell counts – a pilot study

“…This is confirmed by Pipparelli et al. () who showed a higher ECD during organ culture, suggesting that the number of ECs at the time of transplantation is less than surgeons think. Even so, Szaflik et al.…”

Very Early Endothelial Cell Loss After Simultaneous Corneal Autograft and Allograft

“…This very early cell loss is the major factor in the rapid component of the cell loss biphasic curve after a PKP. 7 For PKP with OC corneas, we found a difference of 12% 6 9% (mean 6 SD) between the ebECD and vECD. This ECD decrease reached the cell loss usually admitted at 6 months (228.8% for Bertelmann et al 20 ) and 1 year in normal risk PKP after OC (239.4% for Harper et al, 21 or 229.7% for Borderie et al 3 ).…”

Vulnerability of corneal endothelial cells to mechanical trauma from indentation forces assessed using contact mechanics and fluorescence microscopy

“…However, differing from previous work, we calculated injured CEC densities by acquiring high-resolution fluorescence micrographs (1990×1440 pixels, 905.75 nm/pixel scale) of calcein/PI-stained buttons, counting the number of PI-labelled CEC nuclei and dividing by the contact area. This method contrasts the more common method of staining buttons with trypan blue/alizarin red S and determining the stained area fraction after thresholding (Pipparelli et al, 2011; Saad et al, 2008). This distinct approach was taken for two reasons: first, our main focus was on assessing damage at sites of indentation only; second, the high resolution of our images allowed for reliable visualization and quantification of individual CEC nuclei.…”

Significant differences between specular microscopy and corneal bank endothelial cell counts – a pilot study

“…This is confirmed by Pipparelli et al. () who showed a higher ECD during organ culture, suggesting that the number of ECs at the time of transplantation is less than surgeons think. Even so, Szaflik et al.…”

Very Early Endothelial Cell Loss After Simultaneous Corneal Autograft and Allograft

“…This very early cell loss is the major factor in the rapid component of the cell loss biphasic curve after a PKP. 7 For PKP with OC corneas, we found a difference of 12% 6 9% (mean 6 SD) between the ebECD and vECD. This ECD decrease reached the cell loss usually admitted at 6 months (228.8% for Bertelmann et al 20 ) and 1 year in normal risk PKP after OC (239.4% for Harper et al, 21 or 229.7% for Borderie et al 3 ).…”

Vulnerability of corneal endothelial cells to mechanical trauma from indentation forces assessed using contact mechanics and fluorescence microscopy

“…However, differing from previous work, we calculated injured CEC densities by acquiring high-resolution fluorescence micrographs (1990×1440 pixels, 905.75 nm/pixel scale) of calcein/PI-stained buttons, counting the number of PI-labelled CEC nuclei and dividing by the contact area. This method contrasts the more common method of staining buttons with trypan blue/alizarin red S and determining the stained area fraction after thresholding (Pipparelli et al, 2011; Saad et al, 2008). This distinct approach was taken for two reasons: first, our main focus was on assessing damage at sites of indentation only; second, the high resolution of our images allowed for reliable visualization and quantification of individual CEC nuclei.…”