Regional assessment of energy-producing metabolic activity in the endothelium of donor corneas

Mark A Greiner; Kimberlee A Burckart; Michael D Wagoner; Gregory A Schmidt; Cynthia R Reed; Chase A Liaboe; Miles J Flamme-Wiese; M Bridget Zimmerman; Robert F Mullins; Randy H Kardon; Kenneth M Goins; Benjamin T Aldrich

doi:10.1167/iovs.15-16442

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Regional assessment of energy-producing metabolic activity in the endothelium of donor corneas

Journal article

Peer reviewed

Regional assessment of energy-producing metabolic activity in the endothelium of donor corneas

Mark A Greiner, Kimberlee A Burckart, Michael D Wagoner, Gregory A Schmidt, Cynthia R Reed, Chase A Liaboe, Miles J Flamme-Wiese, M Bridget Zimmerman, Robert F Mullins, Randy H Kardon, …

Investigative ophthalmology & visual science, Vol.56(5), pp.2803-2810

05/2015

DOI: 10.1167/iovs.15-16442

PMID: 26024071

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Abstract

We characterized mitochondrial respiration and glycolysis activity of human corneal endothelium, and compared metabolic activity between central and peripheral regions. Endothelial keratoplasty-suitable corneas were obtained from donors aged 50 to 75 years. The endothelium-Descemet membrane complex (EDM) was isolated, and 3-mm punches were obtained from central and peripheral regions. Endothelium-Descemet membrane punches were assayed for mitochondrial respiration (oxygen consumption) and glycolysis (extracellular acidification) using an extracellular flux analyzer. Enzymatic (citrate synthase, glucose hexokinase) and mitochondrial density (MitoTracker) assays also were performed. Ten corneas were analyzed per assay. Metabolic activity for mitochondrial respiration and glycolysis showed expected changes to assay compounds (P < 0.01, all pairwise comparisons). Basal mitochondrial respiration and glycolysis activity did not differ between regions (P > 0.99). Similarly, central versus peripheral activity after assay compound treatment showed no significant differences (P > 0.99, all time points). The intracorneal coefficient of variation for basal readings between two and four peripheral punches was 18.5% of the mean. Although peripheral samples displayed greater enzymatic activity than central samples (P < 0.05), similar to extracellular flux results, mitochondrial density did not differ between regions (P = 0.78). Extracellular flux analysis of oxygen and pH is a valid technique for characterizing metabolic activity of human corneal endothelium. This technique demonstrates high reproducibility, allows quantification of metabolic parameters using small quantities of live cells, and permits estimation of overall metabolic output. Neither oxygen consumption nor extracellular acidification differed between central and peripheral regions of transplant suitable corneas in this series. Our results show that endothelial cell health can be quantified biochemically in transplant suitable corneas.

Corneal Transplantation

Tissue Culture Techniques

Humans

Middle Aged

Eye Banks

Mitochondria - metabolism

Oxygen Consumption - physiology

Endothelium, Corneal - metabolism

Glycolysis - physiology

Cell Respiration

Aged

Tissue Donors

Energy Metabolism - physiology

Details

Title: Subtitle: Regional assessment of energy-producing metabolic activity in the endothelium of donor corneas
Creators: Mark A Greiner - University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa, United States 2Iowa Lions Eye Bank, Coralville, Iowa, United States
Kimberlee A Burckart - Iowa Lions Eye Bank, Coralville, Iowa, United States
Michael D Wagoner - University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa, United States
Gregory A Schmidt - Iowa Lions Eye Bank, Coralville, Iowa, United States
Cynthia R Reed - Iowa Lions Eye Bank, Coralville, Iowa, United States
Chase A Liaboe - University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa, United States
Miles J Flamme-Wiese - University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa, United States
M Bridget Zimmerman - University of Iowa College of Public Health, Department of Biostatistics, Iowa City, Iowa, United States
Robert F Mullins - University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa, United States
Randy H Kardon - University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa, United States 4Iowa City Veterans Affairs Medical Center and Center of Excellence for the Prevention and Treatment of Visual Loss, Iowa City
Kenneth M Goins - University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa, United States 2Iowa Lions Eye Bank, Coralville, Iowa, United States
Benjamin T Aldrich - University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa, United States 2Iowa Lions Eye Bank, Coralville, Iowa, United States
Resource Type: Journal article
Publication Details: Investigative ophthalmology & visual science, Vol.56(5), pp.2803-2810
DOI: 10.1167/iovs.15-16442
PMID: 26024071
NLM abbreviation: Invest Ophthalmol Vis Sci
ISSN: 0146-0404
eISSN: 1552-5783
Publisher: United States
Language: English
Date published: 05/2015
Academic Unit: Iowa Neuroscience Institute; Biostatistics; Ophthalmology and Visual Sciences
Record Identifier: 9983980083902771

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