Mitochondrial Oxygen Consumption by the Foreskin and its Fibroblast-rich Culture

Abstract

Objectives: This study investigated the feasibility of using a phosphorescence oxygen analyser to measure cellular respiration (mitochondrial O₂ consumption) in foreskin samples and their fibroblast-rich cultures.Methods: Foreskin specimens from normal infants were collected immediately after circumcision and processed for measuring cellular respiration and for culture. Cellular mitochondrial O₂ consumption was determined as a function of time from the phosphorescence decay of the Pd (II) meso-tetra-(4-sulfonatophenyl)-tetrabenzoporphyrin. Results: In sealed vials containing a foreskin specimen and glucose, O₂ concentration decreased linearly with time, confirming the zero-order kinetics of O₂ consumption by cytochrome oxidase. Cyanide inhibited O₂ consumption, confirming that the oxidation occurred mainly in the mitochondrial respiratory chain. The rate of foreskin respiration (mean ± SD) was 0.074 ± 0.02 μM O₂ min^-1 mg^-1 (n = 23). The corresponding rate for fibroblast-rich cultures was 9.84 ± 2.43 μM O₂ min^-1 per 10⁷ cells (n = 15). Fibroblast respiration was significantly lower in a male infant with dihydrolipoamide dehydrogenase gene mutations, but normalised with the addition of thiamine or carnitine. Conclusion: The foreskin and its fibroblast-rich culture are suitable for assessment of cellular respiration. However, the clinical utility of foreskin specimens to detect disorders of impaired cellular bioenergetics requires further investigation.