September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
The Endothelial Safety of Using Preloaded Descemet Membrane Endothelial Keratoplasty Graft
Author Affiliations & Notes
  • Meraf Amde Wolle
    Kellogg Eye Center, University of Michigan, Ann Arbor, Michigan, United States
  • David DeMill
    Kellogg Eye Center, University of Michigan, Ann Arbor, Michigan, United States
  • Maria A Woodward
    Kellogg Eye Center, University of Michigan, Ann Arbor, Michigan, United States
  • Lauren Johnson
    Eversight Michigan, Ann Arbor, Michigan, United States
  • Shahzad Mian
    Kellogg Eye Center, University of Michigan, Ann Arbor, Michigan, United States
  • Footnotes
    Commercial Relationships   Meraf Wolle, None; David DeMill, None; Maria Woodward, None; Lauren Johnson, None; Shahzad Mian, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 1244. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Meraf Amde Wolle, David DeMill, Maria A Woodward, Lauren Johnson, Shahzad Mian; The Endothelial Safety of Using Preloaded Descemet Membrane Endothelial Keratoplasty Graft. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1244.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : Descemet Membrane Endothelial Keratoplasty (DMEK) surgeries are on the rise and ways to streamline the procedure, such as preloading tissue, would assist in increasing surgeon uptake. The goal of this study is to determine the safety of preloading DMEK grafts 24 hours prior to injection by analyzing endothelial cell loss (ECL) using commercially available software Metamorph and comparing it to the previously validated software Fiji.

Methods : Eighteen 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, as controls. The remaining nine corneas were preloaded in the injector for 24 hours prior to injection. The stained corneas were imaged using an inverted fluorescent microscope (Fig 1). ECL was then analyzed and quantified by 2 different graders using Metamorph and Fiji (Fig 2). The study was powered to detect a 10% difference in cell loss between the two groups.

Results : The control DMEK tissue resulted in 26.29%±9.58% ECL and the tissue stored for 24 hours prior to injection resulted in 22.60%±8.72% ECL (p=0.42). The mean (SD) pooled ECL as analyzed by Metamorph was 23.70% ±8.40% and by Fiji was 26.30%±7.55%. Interobserver agreement was 0.97 (0.87-0.99) for MetaMorph and 0.96 (0.84-0.99) for Fiji. The average time required for image analysis was 2 min for MetaMorph and 15 minutes for Fiji. Intraobserver agreement was 0.99 (0.94-1.00) for MetaMorph and 0.96 (0.82-0.99) for Fiji.

Conclusions : Preloading DMEK tissue in an injector does not result in significantly increased endothelial cell loss showing the clinical feasibility of such an approach. Clinical trials are needed to draw conclusions about clinical viability. Metamorph is a novel and accurate imaging software with increased efficiency and reproducibility in quantifying ECL as compared to the previously validated Fiji. Improving analysis techniques will have long lasting consequences on future experiments on endothelial graft preparation and surgical technique.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

 

Fig 1. A-Vital dye stained cornea image captured initially in 3D. B-Cross sections through the adjacent full cornea image showing the undulations of the tissue on the z axis. C-Overlay of the intensity of fluorescence at a given location.

Fig 1. A-Vital dye stained cornea image captured initially in 3D. B-Cross sections through the adjacent full cornea image showing the undulations of the tissue on the z axis. C-Overlay of the intensity of fluorescence at a given location.

 

Fig 2. MetaMorph (left) and Fiji (right) final images after analysis showing ECL (hyperfluorescent or black areas).

Fig 2. MetaMorph (left) and Fiji (right) final images after analysis showing ECL (hyperfluorescent or black areas).

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×