July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Metabolic Optimization of Endothelial Keratoplasty Graft Tissue with Coenzyme Q10
Author Affiliations & Notes
  • Jessica Marie Skeie
    University of Iowa, Coralville, Iowa, United States
    Iowa Lions Eye Bank, Iowa, United States
  • Benjamin T Aldrich
    University of Iowa, Coralville, Iowa, United States
    Iowa Lions Eye Bank, Iowa, United States
  • Anna E Wilcox
    University of Iowa, Coralville, Iowa, United States
    Iowa Lions Eye Bank, Iowa, United States
  • Gregory A Schmidt
    University of Iowa, Coralville, Iowa, United States
    Iowa Lions Eye Bank, Iowa, United States
  • Cynthia R Reed
    University of Iowa, Coralville, Iowa, United States
    Iowa Lions Eye Bank, Iowa, United States
  • Mark A Greiner
    University of Iowa, Coralville, Iowa, United States
    Iowa Lions Eye Bank, Iowa, United States
  • Footnotes
    Commercial Relationships   Jessica Skeie, None; Benjamin Aldrich, None; Anna Wilcox, None; Gregory Schmidt, None; Cynthia Reed, None; Mark Greiner, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 3823. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Jessica Marie Skeie, Benjamin T Aldrich, Anna E Wilcox, Gregory A Schmidt, Cynthia R Reed, Mark A Greiner; Metabolic Optimization of Endothelial Keratoplasty Graft Tissue with Coenzyme Q10. Invest. Ophthalmol. Vis. Sci. 2019;60(9):3823.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : To determine if adding the antioxidant Coenzyme Q10 (CoQ10 or ubiquinol) to donor cornea storage media enhances the metabolic function of corneal endothelial cells (CECs) and/or decreases overall cell death in storage.

Methods : Human corneal tissue pairs were obtained by Iowa Lions Eye Bank (ILEB) from nondiabetic donors 60-75 years old and stored in Optisol GS (Bausch + Lomb, Irvine, CA) at 4°C following procurement in accordance with Eye Bank Association of America and ILEB policies and procedures. For 5 days prior to testing, but within 9 days of procurement, one stored tissue from a corneal pair was treated with 10 µg/mL CoQ10, while the mate tissue was treated with diluent only as a control. Descemet membrane and endothelial cell punches were collected and mounted onto the bottom of a Seahorse assay plate (cells facing upward). Mitochondrial respiration was assayed by measuring oxygen consumption using the Seahorse XFe24 Extracellular Flux Analyzer (Agilent Technologies, Santa Clara, CA) over 120 minutes (9-minute intervals). Punches were labeled with a nuclear counterstain (DAPI) and remaining tissues were mounted onto slides and labeled with 488A-Annexin V, Ethidium Homodimer III, and Hoechst 33342 (Apoptotic, Necrotic, and Healthy Cells Quantification Kit; Biotium, Fremont, CA) to assay cell health. Nuclei were counted in each punch to normalize respirometry data. Immunohistochemistry densitometry was measured using ImageJ software.

Results : In total, 14 paired corneas were tested. Three different aspects of mitochondrial respiration were affected by CoQ10 treatment: proton leak was increased 34% (p=0.046), maximal respiration was increased 97% (p=0.003), and spare respiratory capacity was increased 133% (p<0.001). Corneal endothelial cell necrosis was not changed, however, apoptosis was reduced 29% in treated cells (p=0.09).

Conclusions : In this series, Coenzyme Q10 increased corneal endothelial cell mitochondrial respiration and prevented cells from dying in storage. Findings indicate that Optisol GS supplemented with CoQ10 may reduce presurgical cell death and functional decline related to tissue storage. Further studies are needed to optimize the dosing strategy during storage as well as to determine the cytoprotective effects on cell density after endothelial keratoplasty.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

×
×

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.

×