Electron microscopic study of the endothelium of stored and cryopreserved monkey corneas

  • Wolfgang Waller
  • Diane L. Van Horn
Article

Summary

Monkey corneas were either cryopreserved at a controlled rate over liquid nitrogen in a solution containing dimethylsulfoxide, sucrose and serum albumin, or were stored in moist chambers at 4°C for 48 hours. The endothelium was examined by scanning and transmission electron microscopy and the results compared. The entire endothelial layer of cells was always intact and there was little evidence of freeze-thaw-induced structural damage in the cryopreserved corneas. Mitochondrial swelling and clumping of nuclear chromatin was observed in the stored corneas.

Keywords

Nitrogen Microscopy Electron Microscopy Sucrose Transmission Electron Microscopy 

Zusammenfassung

Hornhäute von Rhesusaffen (Macaca arctoides) wurden entweder nach der Methode von Kaufman und Capella gefrierkonserviert oder in feuchten Kammern bei +4°C für 48 Std gelagert. Anschließend wurde das Endothel elektronenmikroskopisch untersucht. Die vergleichende Auswertung ergab, daß die gesamte endotheliale Zellschicht immer intakt war. Die gefrierkonservierten Hornhäute waren in ihrer Struktur durch den Gefrierprozeß nur geringfügig geschädigt. In den bei + 4° C gelagerten Hornhäuten waren Schwellung der Mitochondrien und Verklumpung des Kernchromatins zu beobachten.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Capella, J. A., Kaufman, H. E., Robbins, J. E.: Preservation of viable corneal tissue. Arch. Ophthal. 74, 669 (1965).Google Scholar
  2. Cleveland, P. H., Schneider, C. W.: A simple method of preserving tissue for scanning electron microscopy. Vision Res. 9 1401 (1969).Google Scholar
  3. Eastcott, H. H. G., Cross, A. G., Leigh, A. G., North, D. P.: Preservation of corneal grafts by freezing. Lancet 1954I, 237.Google Scholar
  4. Horn, D. L. Van, Edelhauser, H. F., Gallun, A. B., Schultz, R. O.: Reversibility of freeze-thaw-induced injury. Arch. Ophthal. 87, 422 (1972).Google Scholar
  5. Horn, D. L. Van, Hanna, C., Schultz, R. O.: Corneal cryopreservation. II. Ultrastructural and viability changes. Arch. Ophthal. 84, 655 (1970).Google Scholar
  6. Horn, D. L. Van, Schultz, R. O.: (a) Ultrastructural changes in the endothelium of human corneas stored under eye-bank conditions, p. 29. (b) Ultrastructural changes in cryopreserved and experimentally rehydrated corneal tissue, p. 237. In: Corneal Preservation, ed. by Capella, J. A., Edelhauser, H. F., Horn, D. L. Van. Springfield: C. C. Thomas 1972.Google Scholar
  7. Iliff, C. E., Wood, R. M., Hollander, D. H.: Successful transplantation of a frozen human cornea. Amer. J. Ophthal. 41, 505 (1956).Google Scholar
  8. Katzin, H. M.: The preservation of corneal tissue by freezing and dehydration. Amer. J. Ophthal. 30, 1128 (1947).Google Scholar
  9. Kaufman, H. E., Capella, J. A.: Preserved corneal tissue for transplantation. J. Crysosurg. 1, 125 (1968).Google Scholar
  10. Kaufman, H. E., Escapini, H., Capella, J. A., Robbins, J. E., Kaplan, M.: Living preserved corneal tissue for penetrating keratoplasty. Arch. Ophthal. 76, 471 (1966).Google Scholar
  11. Leopold, I. H., Adler, F. H.: Use of frozen-dried cornea as transplant material. Arch. Ophthal. 37, 268 (1947).Google Scholar
  12. Lovelock, J. E., Bishop, M. W. H.: Prevention of freezing damage to living cells by dimethyl sulphoxide. Nature (Lond.) 183, 1394 (1959).Google Scholar
  13. Mathieu, M.: Results obtained in perforating keratoplasty with preserved corneas, using liquid nitrogen. Trans. Amer. Acad. Ophthal. Otolaryng. 74, 399 (1970).Google Scholar
  14. Mueller, F. O., Casey, T. A., Trevor-Roper, P. D.: Use of deep-frozen human cornea in full thickness grafts. Brit. med. J. 1964II, 473.Google Scholar
  15. Mueller, F. O., Smith, A. U.: Some experiments on grafting frozen corneal tissue in rabbits. Exp. Eye Res. 2, 237 (1963).Google Scholar
  16. Polge, C., Smith, A. U., Parks, A. S.: Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature (Lond.) 164, 666 (1949).Google Scholar
  17. Rycroft, B. W.: Three unusual corneal grafts. Brit. J. Ophthal. 41, 759 (1957).Google Scholar
  18. Schultz, R. O.: Laboratory evaluation of cryopreserved corneal tissue. Trans. Amer. ophthal. Soc. 69, 563 (1971).Google Scholar
  19. Sherman, J. K.: Freeze-thaw-induced structural changes in cells. II. Examples of cryoinjury of cellular structure. J. Cryosurg. 2, 156 (1969).Google Scholar
  20. Smelser, G. K., Ozanics, V.: Effect of quick freezing at very low temperatures of donor tissue in corneal transplants. Proc. Soc. exp. Biol. (N.Y.) 62, 274 (1946).Google Scholar
  21. Smith, A. U., Ashwood-Smith, M. J., Young, M. R.: Some in vitro studies on rabbit corneal tissue. Exp. Eye Res. 2, 71 (1963).Google Scholar
  22. Svedbergh, B., Bill, A.: Scanning electron microscopic studies of the corneal endothelium in man and monkeys. Acta ophthal. (Kbh.) 50, 321 (1972).Google Scholar

Copyright information

© Springer-Verlag 1973

Authors and Affiliations

  • Wolfgang Waller
    • 1
  • Diane L. Van Horn
    • 2
  1. 1.Augenklinik der Universität WürzburgDeutschland
  2. 2.Departments of Ophthalmology and Physiology, Medical College of Wisconsin and Research ServiceVeterans Administration CenterWood (Milwaukee)

Personalised recommendations