Quantitative Analysis of Endothelial Cell Loss in Preloaded Descemet Membrane Endothelial Keratoplasty Grafts

Meraf Wolle, David L. Demill, Lauren Johnson, Stephen I. Lentz, Maria A. Woodward, Shahzad I. Mian

Research output: Contribution to journalArticle

Abstract

Purpose: Availability of preloaded Descemet membrane endothelial keratoplasty (pDMEK) tissue may increase acceptance of DMEK in surgical management of endothelial disease. The goal of this study was to determine the safety of pDMEK grafts for 24 hours before surgery by analyzing endothelial cell loss (ECL) using 2 image analysis software programs. Methods: A total of 18 cadaveric corneas were prepared for DMEK using a standardized technique and loaded in a modified Jones tube injector. Nine of the corneas were injected into Calcein AM vital dye after 1 minute (controls), and the remaining 9 corneas were left preloaded for 24 hours before injection into vital dye for staining. The stained corneas were imaged using an inverted confocal microscope. ECL was then analyzed and quantified by 2 different graders using 2 image analysis software programs. Results: The control DMEK tissue resulted in 22.0% ± 4.0% ECL compared with pDMEK tissue, which resulted in 19.2% ± 7.2% ECL (P = 0.31). Interobserver agreement was 0.93 for MetaMorph and 0.92 for Fiji. The average time required to process images with MetaMorph was 2 ± 1 minutes and with Fiji was 20 ± 10 minutes. Intraobserver agreement was 0.97 for MetaMorph and 0.93 for Fiji. Conclusions: Preloading DMEK tissue is safe and may provide an alternative technique for tissue distribution and surgery for DMEK. The use of MetaMorph software for quantifying ECL is a novel and accurate imaging method with increased efficiency and reproducibility compared with the previously validated Fiji.

Original languageEnglish (US)
Pages (from-to)1295-1301
Number of pages7
JournalCornea
Volume36
Issue number11
DOIs
StatePublished - Nov 1 2017

Fingerprint

Descemet Membrane
Fiji
Corneal Transplantation
Endothelial Cells
Cornea
Transplants
Software
Endothelium
Coloring Agents
Membranes
Tissue Distribution
Disease Management
Staining and Labeling
Safety
Injections

Keywords

  • DMEK
  • endothelial cell loss
  • preloading
  • vital dye staining

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Quantitative Analysis of Endothelial Cell Loss in Preloaded Descemet Membrane Endothelial Keratoplasty Grafts. / Wolle, Meraf; Demill, David L.; Johnson, Lauren; Lentz, Stephen I.; Woodward, Maria A.; Mian, Shahzad I.

In: Cornea, Vol. 36, No. 11, 01.11.2017, p. 1295-1301.

Research output: Contribution to journalArticle

Wolle, Meraf ; Demill, David L. ; Johnson, Lauren ; Lentz, Stephen I. ; Woodward, Maria A. ; Mian, Shahzad I. / Quantitative Analysis of Endothelial Cell Loss in Preloaded Descemet Membrane Endothelial Keratoplasty Grafts. In: Cornea. 2017 ; Vol. 36, No. 11. pp. 1295-1301.
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N2 - Purpose: Availability of preloaded Descemet membrane endothelial keratoplasty (pDMEK) tissue may increase acceptance of DMEK in surgical management of endothelial disease. The goal of this study was to determine the safety of pDMEK grafts for 24 hours before surgery by analyzing endothelial cell loss (ECL) using 2 image analysis software programs. Methods: A total of 18 cadaveric corneas were prepared for DMEK using a standardized technique and loaded in a modified Jones tube injector. Nine of the corneas were injected into Calcein AM vital dye after 1 minute (controls), and the remaining 9 corneas were left preloaded for 24 hours before injection into vital dye for staining. The stained corneas were imaged using an inverted confocal microscope. ECL was then analyzed and quantified by 2 different graders using 2 image analysis software programs. Results: The control DMEK tissue resulted in 22.0% ± 4.0% ECL compared with pDMEK tissue, which resulted in 19.2% ± 7.2% ECL (P = 0.31). Interobserver agreement was 0.93 for MetaMorph and 0.92 for Fiji. The average time required to process images with MetaMorph was 2 ± 1 minutes and with Fiji was 20 ± 10 minutes. Intraobserver agreement was 0.97 for MetaMorph and 0.93 for Fiji. Conclusions: Preloading DMEK tissue is safe and may provide an alternative technique for tissue distribution and surgery for DMEK. The use of MetaMorph software for quantifying ECL is a novel and accurate imaging method with increased efficiency and reproducibility compared with the previously validated Fiji.

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