The effect of hEGF and insulin on corneal metabolism during Optisol storage.
We examined the effect of the growth factors, human epidermal growth factor (hEGF) and insulin, on corneal metabolism during storage in Optisol, a chondroitin-sulfate-(CS)-based storage medium. Paired cat corneas, in either Optisol only or Optisol with growth factor(s), were analyzed using ex vivo 31P nuclear magnetic resonance, after storage for 1 week at 4 degrees C. Lysosomal enzyme release into the media at the end of the storage period also was measured fluorometrically. Both epithelial-intact and epithelial-denuded corneal pairs were examined for all conditions. Considering corneas having either intact epithelia or epithelium-denuded corneas, the addition of either growth factor alone to Optisol did not alter the relative corneal concentrations of five of the six phosphatic metabolite spectral bands measured or two metabolic ratios calculated from these bands. Phosphodiesters, however, were significantly lower in corneas stored in Optisol containing both hEGF and insulin (23%) than in corneas stored in Optisol alone (30%). Intracorneal pH was unaffected by the addition of growth factor(s). A significantly higher release of alpha-glucosidase and alpha-mannosidase was noted in those corneas stored in Optisol containing both hEGF and insulin. Optisol maintains high-energy phosphate corneal metabolism similar to other CS-based media, K-Sol and Chondroitin Sulfate Corneal Storage Medium (CSM). The addition of the growth factors hEGF and insulin to Optisol alters corneal metabolic activity during storage in a manner indicative of conserving corneal phospholipids.
Lass
J H
JH
Department of Ophthalmology, Case Western Reserve University, Cleveland, Ohio.
Putman
S C
SC
Bruner
W E
WE
Cano
D B
DB
Greiner
J V
JV
Glonek
T
T
D000818:Animals; D002415:Cats; D002809:Chondroitin Sulfates; D045424:Complex Mixtures; D003315:Cornea / Q000187:drug effects / Q000378:metabolism*; D015925:Cryopreservation; D016895:Culture Media, Serum-Free; D003911:Dextrans; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D004848:Epithelium; D005839:Gentamicins; D006026:Glycoside Hydrolases / Q000378:metabolism; D007328:Insulin / Q000494:pharmacology*; D009682:Magnetic Resonance Spectroscopy; D010710:Phosphates / Q000378:metabolism; D010758:Phosphorus; D014021:Tissue Preservation
D045424:Complex Mixtures; D016895:Culture Media, Serum-Free; D003911:Dextrans; D005839:Gentamicins; D007328:Insulin; D010710:Phosphates; D010758:Phosphorus; D004815:Epidermal Growth Factor; D002809:Chondroitin Sulfates; C075736:Optisol; D006026:Glycoside Hydrolases