Glutathione preservation during storage of rat lenses in optisol-GS and castor oil

PLoS One. 2013 Nov 19;8(11):e79620. doi: 10.1371/journal.pone.0079620. eCollection 2013.

Abstract

Background: Glutathione concentration in the lens decreases in aging and cataractous lenses, providing a marker for tissue condition. Experimental procedures requiring unfrozen lenses from donor banks rely on transportation in storage medium, affecting lens homeostasis and alterations in glutathione levels. The aim of the study was to examine the effects of Optisol-GS and castor oil on lens condition, determined from their ability to maintain glutathione concentrations.

Methodology/principal findings: Rat lenses were stored in the two types of storage media at varying time intervals up to 3 days. Glutathione concentration was afterwards determined in an enzymatic detection assay, specific for both reduced and oxidized forms. Lenses removed immediately after death exhibited a glutathione concentration of 4.70±0.29 mM. In vitro stored lenses in Optisol-GS lost glutathione quickly, ending with a concentration of 0.60±0.34 mM after 3 days while castor oil stored lenses exhibited a slower decline and ended at 3 times the concentration. A group of lenses were additionally stored under post mortem conditions within the host for 6 hours before its removal. Total glutathione after 6 hours was similar to that of lenses removed immediately after death, but with altered GSH and GSSG concentrations. Subsequent storage of these lenses in media showed changes similar to those in the first series of experiments, albeit to a lesser degree.

Conclusions/significance: It was determined that storage in Optisol-GS resulted in a higher loss of glutathione than lenses stored in castor oil. Storage for more than 12 hours reduced glutathione to half its original concentration, and was considered unusable after 24 hours.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Castor Oil / chemistry*
  • Chondroitin Sulfates / chemistry*
  • Complex Mixtures / chemistry
  • Dextrans / chemistry*
  • Gentamicins / chemistry*
  • Glutathione / chemistry*
  • In Vitro Techniques
  • Lens, Crystalline*
  • Male
  • Organ Preservation / methods*
  • Organ Preservation Solutions / chemistry
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Complex Mixtures
  • Dextrans
  • Gentamicins
  • Organ Preservation Solutions
  • Castor Oil
  • Chondroitin Sulfates
  • Optisol
  • Glutathione

Grant support

Funding statement: The study was funded by the Danish Medical Research Council (http://fivu.dk/forskning-og-innovation/rad-og-udvalg/det-frie-forskningsrad/radet/dff-sundhed-og-sygdom), The Foundation of June 15 http://www.15junifonden.dk/content/dk/om_15_juni_fonden), Manufacturer Einar Willumsens memorial scholarship and merchant Chr. Andersen and wife Ingeborg Andersen Scholarship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.