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