Hypothermic preservation of corneas in a hyperkalaemic solution (CPTES): II. Extended storage in the presence of chondroitin sulphate.
Periods of preservation for donor corneas, even for short times, are necessary to facilitate optimum conditions in penetrating keratoplasty. However, current techniques for corneal storage at low temperatures may not provide optimal conditions for maintaining tissue integrity. In particular, the ionic composition of the storage medium has received little attention since it has been assumed throughout that the normal complement of ions in tissue culture media will also be suitable for preservation at reduced temperatures. This study extends our previous investigations on the merits of using CPTES (corneal-potassium-TES), a potassium-rich balanced salt solution containing an impermeant anionic pH buffer (TES), as a storage solution specifically designed to prevent the loss of intracellular potassium and minimise endothelial cell swelling during the time that the normal regulatory processes are switched off. The effect of adding the natural polymer chondroitin sulphate (CS) as a colloid osmotic agent to the hyperkalaemic storage medium is now examined. Corneas stored in CPTES containing 2.5% chondroitin sulphate retained a very high level of structural and functional integrity after three, five, and seven days storage at 0 degrees C; furthermore, stromal swelling was restricted to only 21%. All corneas stored in CPTES + 2.5% CS showed active endothelial function by thinning efficiently at rates that were greater than those previously reported for rabbit corneas stored for similar lengths of time in either M-K medium or K-sol. The zwitterionic buffers TES and HEPES were interchangeable in the hyperkalaemic solution and were non-toxic to corneal endothelium at a concentration of 100 mM. These compounds offer excellent pH buffering in bicarbonate-free medium.
Taylor
M J
MJ
MRC Medical Cryobiology Group, Cambridge.
Hunt
C J
CJ
Madden
P W
PW
D000818:Animals; D002021:Buffers; D002807:Chondroitin; D002809:Chondroitin Sulfates; D003315:Cornea / Q000033:anatomy & histology; D004728:Endothelium, Corneal / Q000166:cytology; D005615:Freezing; D006982:Hypertonic Solutions; D011188:Potassium; D011817:Rabbits; D013997:Time Factors; D014021:Tissue Preservation / Q000379:methods*; D014325:Tromethamine / Q000031:analogs & derivatives
D002021:Buffers; D006982:Hypertonic Solutions; D014325:Tromethamine; C004551:TES; D002807:Chondroitin; D002809:Chondroitin Sulfates; D011188:Potassium