Skip to main content
PMC home page
The British Journal of Ophthalmology logoLink to The British Journal of Ophthalmology
. 1989 Oct;73(10):781–791. doi: 10.1136/bjo.73.10.781

Hypothermic preservation of corneas in a hyperkalaemic solution (CPTES): I. Short-term storage in the absence of colloid osmotic agents.

M J Taylor 1, C J Hunt 1
PMCID: PMC1041887  PMID: 2818988

Abstract

Preservation solutions for short-term storage of isolated donor corneas for use in penetrating keratoplasty have all been based on tissue culture medium, on the assumption that media designed to maintain the viability of cells at physiological temperatures will also provide suitable conditions for preservation at reduced temperatures. But for hypothermic preservation of some other tissues and organs, when ionic pumps are inhibited, it is unnecessary to support metabolism, and beneficial control of ion and water distribution between intra- and extracellular compartments is achieved by storage in appropriately formulated 'intracellular-type' solutions. We have therefore designed a solution that will restrict ionic imbalances and minimise endothelial cell swelling in corneas during exposure at reduced temperatures. This potassium-rich solution contains the biological pH buffer TES as an impermeant anion and is designated CPTES (corneal-potassium-TES). The structural and functional integrity of rabbit corneas stored at 0 degrees C in CPTES, without the addition of colloid osmotic agents, is compared with that of corneas stored in glutathione bicarbonate Ringers' solution (GBR), an 'extracellular-type' medium formulated for the maintenance of endothelial integrity during in-vitro perfusion at 34 degrees C. Corneas swelled significantly less during storage in CPTES than in GBR and could be stored for five days before reaching the same degree of hydration as corneas stored for only three days in GBR. Gross structural integrity and endothelial ultrastructure were maintained during storage for three and five days in CPTES. The rate of thinning of corneas stored in CPTES was significantly greater than in comparable groups of corneas stored in GBR. However, the efficient dehydration of corneas stored in CPTES was always preceded during perfusion by a brief period of additional swelling which was shown to be an osmotic response during the elution of the buffer compound TES that had permeated the stroma during storage. The omission of calcium or the addition of adenosine and glutathione to the CPTES preservation medium had no detectable effect on the integrity of the endothelium, but the omission of bicarbonate was beneficial, producing significantly higher rates of stromal thinning during normothermic perfusion. Additional benefits for extending storage by including colloid osmotic agents are described in a companion paper.

Full text

PDF
782

Images in this article

785Image
on p.785
785Image
on p.785

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bourne W. M. Endothelial cell survival on transplanted human corneas preserved at 4 C in 2.5% chondroitin sulfate for one to 13 days. Am J Ophthalmol. 1986 Sep 15;102(3):382–386. doi: 10.1016/0002-9394(86)90015-2. [DOI] [PubMed] [Google Scholar]
  2. Dikstein S., Maurice D. M. The metabolic basis to the fluid pump in the cornea. J Physiol. 1972 Feb;221(1):29–41. doi: 10.1113/jphysiol.1972.sp009736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Edelhauser H. F., Van Horn D. L., Hyndiuk R. A., Schultz R. O. Intraocular irrigating solutions. Their effect on the corneal endothelium. Arch Ophthalmol. 1975 Aug;93(8):648–657. doi: 10.1001/archopht.1975.01010020614011. [DOI] [PubMed] [Google Scholar]
  4. Garlick P. B., Radda G. K., Seeley P. J. Studies of acidosis in the ischaemic heart by phosphorus nuclear magnetic resonance. Biochem J. 1979 Dec 15;184(3):547–554. doi: 10.1042/bj1840547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Good N. E., Winget G. D., Winter W., Connolly T. N., Izawa S., Singh R. M. Hydrogen ion buffers for biological research. Biochemistry. 1966 Feb;5(2):467–477. doi: 10.1021/bi00866a011. [DOI] [PubMed] [Google Scholar]
  6. Hodson S. The endothelial pump of the cornea. Invest Ophthalmol Vis Sci. 1977 Jul;16(7):589–591. [PubMed] [Google Scholar]
  7. Hull D. S., Green K., Bowman K., Csukas S., Riley M. V. Intracellular pH and glutathione levels in rabbit corneal endothelium following storage in moist chamber and MK medium. Invest Ophthalmol Vis Sci. 1983 Feb;24(2):214–217. [PubMed] [Google Scholar]
  8. KINSEY V. E. The chemical composition and the osmotic pressure of the aqueous humor and plasma of the rabbit. J Gen Physiol. 1951 Jan;34(3):389–402. doi: 10.1085/jgp.34.3.389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaufman H. E., Varnell E. D., Kaufman S. Chondroitin sulfate in a new cornea preservation medium. Am J Ophthalmol. 1984 Jul 15;98(1):112–114. doi: 10.1016/0002-9394(84)90198-3. [DOI] [PubMed] [Google Scholar]
  10. Mayes K. R., Graham M. V., Hodson S. The deleterious effects of exogenous bicarbonate on the rabbit cornea undergoing prolonged refrigerated storage. Exp Eye Res. 1978 May;26(5):555–560. doi: 10.1016/0014-4835(78)90065-9. [DOI] [PubMed] [Google Scholar]
  11. McCarey B. E., Kaufman H. E. Improved corneal storage. Invest Ophthalmol. 1974 Mar;13(3):165–173. [PubMed] [Google Scholar]
  12. Schimmelpfennig B. Corneal preservation at -4 degrees C in a modified tissue culture medium. Albrecht Von Graefes Arch Klin Exp Ophthalmol. 1978 Apr 7;206(1):57–66. doi: 10.1007/BF00411338. [DOI] [PubMed] [Google Scholar]
  13. Sherrard E. S. An application of the temperature reversal effect: a possible method to evaluate donor corneae for penetrating keratoplasty. Exp Eye Res. 1973 May 10;15(5):667–675. doi: 10.1016/0014-4835(73)90075-4. [DOI] [PubMed] [Google Scholar]
  14. Sherrard E. S. Method of evaluating donor corneae for transplantation. Br J Ophthalmol. 1973 Apr;57(4):244–252. doi: 10.1136/bjo.57.4.244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Taylor M. J. Clinical cryobiology of tissues: preservation of corneas. Cryobiology. 1986 Aug;23(4):323–353. doi: 10.1016/0011-2240(86)90038-6. [DOI] [PubMed] [Google Scholar]
  16. Taylor M. J., Hunt C. J. A new preservation solution for storage of corneas at low temperatures. Curr Eye Res. 1985 Sep;4(9):963–973. doi: 10.3109/02713689509000003. [DOI] [PubMed] [Google Scholar]
  17. Taylor M. J., Hunt C. J. Dual staining of corneal endothelium with trypan blue and alizarin red S: importance of pH for the dye-lake reaction. Br J Ophthalmol. 1981 Dec;65(12):815–819. doi: 10.1136/bjo.65.12.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Taylor M. J., Hunt C. J., Madden P. W. Hypothermic preservation of corneas in a hyperkalaemic solution (CPTES): II. Extended storage in the presence of chondroitin sulphate. Br J Ophthalmol. 1989 Oct;73(10):792–802. doi: 10.1136/bjo.73.10.792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Taylor M. J., Hunt C. J. Tolerance of corneas to multimolar dimethyl sulfoxide at 0 degrees C. Implications for cryopreservation. Invest Ophthalmol Vis Sci. 1989 Mar;30(3):400–412. [PubMed] [Google Scholar]
  20. Taylor M. J., Pignat Y. Practical acid dissociation constants, temperature coefficients, and buffer capacities for some biological buffers in solutions containing dimethyl sulfoxide between 25 and -12 degrees C. Cryobiology. 1982 Feb;19(1):99–109. doi: 10.1016/0011-2240(82)90129-8. [DOI] [PubMed] [Google Scholar]
  21. Taylor M. J. The role of pH and buffer capacity in the recovery of function of smooth muscle cooled to -13 degrees C in unfrozen media. Cryobiology. 1982 Dec;19(6):585–601. doi: 10.1016/0011-2240(82)90188-2. [DOI] [PubMed] [Google Scholar]

Articles from The British Journal of Ophthalmology are provided here courtesy of BMJ Publishing Group

RESOURCES