Abstract
Purpose
To evaluate temporary exposure to hypothermia for its effects on endothelial cell density of porcine corneas in dextran containing organ-culture medium, with regard to possible negative influences of low temperatures during the transport of corneal grafts.
Methods
Two groups of central discs from pig corneas (diameter 8 mm) were first organ-cultured (MEM with 6% dextran 500) for 24 hours at 32°C. Twelve corneas were exposed to 4°C in group 1 for 12 hours and to 21°C in group 2 for 48 hours each. The paired corneal discs were not treated, and served as controls. After further organ culture of all corneas for 48 hours at 32°C to allow regenerative processes, corneal endothelium was stained with alizarin red S and examined by light microscopy. The endothelial cell densities were determined manually on three central images.
Results
Exposure for 12 hours to 4°C as well as for 48 hours to 21°C induced an endothelial cell loss of 0.3% and 1.8% respectively. Statistical analysis showed no significant difference (p = 0.680) of the endothelial cell density between corneas exposed to 4°C and the control corneas (4166 ± 389 cells/mm2 and 4177 ± 407 cells/mm2 respectively). Despite the minor cell loss, the difference of the endothelial cell density between corneas exposed to 21°C and the control corneas (4085 ± 260 cells/mm2 and 4159 ± 312 cells/mm2 respectively) was statistically significant (p = 0.025).
Conclusions
Exposure of organ-cultured porcine corneas in dextran containing medium to 4°C for 12 hours and 21°C for 48 hours does not compromise the endothelial cell density of donor corneas in a clinically relevant manner. A storage of corneal grafts at temperatures down to 4°C for 12 hours, as might be the case during transport from the cornea bank to the ocular surgeon, does not seem to damage the endothelial cell layer.
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References
Commission Directive 2006/17/EC. In: Official Journal of the European Union; 2006. p 40–52
Armitage WJ, Easty DL (1997) Factors influencing the suitability of organ-cultured corneas for transplantation. Invest Ophthalmol Vis Sci 38:16–24
Bahn CF, Glassman RM, MacCallum DK, Lillie JH, Meyer RF, Robinson BJ et al (1986) Postnatal development of corneal endothelium. Invest Ophthalmol Vis Sci 27:44–51
Camposampiero D, Tiso R, Zanetti E, Ruzza A, Bruni A, Ponzin D (2003) Improvement of human corneal endothelium in culture after prolonged hypothermic storage. Eur J Ophthalmol 13:745–751
Hagenah M, Bohnke M (1993) Latent endothelial cell damage after experimental corneal cryopreservation. Graefes Arch Clin Exp Ophthalmol 231:529–532
Hu FR, Tsai AC, Wang IJ, Chang SW (1999) Outcomes of penetrating keratoplasty with imported donor corneas. Cornea 18:182–187
Jacobsen U, Michels G, Liedtke G, Muller MC, Reim M (1985) Organ-culture of pig cornea - methods, clinical morphology and histology. Ophthalmic Res 17:201
Komuro A, Hodge DO, Gores GJ, Bourne WM (1999) Cell death during corneal storage at 4 degrees C. Invest Ophthalmol Vis Sci 40:2827–2832
Lindstrom RL, Doughman DJ, Vanhorn DL, Schmitt MK, Byrnes PJ (1977) Organ-culture corneal storage at ambient room-temperature. Arch Ophthalmol 95:869–878
Lindstrom RL, Kaufman HE, Skelnik DL, Laing RA, Lass JH, Musch DC et al (1992) Optisol corneal storage medium. Am J Ophthalmol 114:345–356
Madden PW, Maguire SH (1996) Methods of refrigerated transport of corneas for transplantation. Aust N Z J Ophthalmol 24:17–20
Meltendorf C, Ohrloff C, Rieck P, Schroeter J (2007) Endothelial cell density in porcine corneas after exposure to hypotonic solutions. Graefes Arch Clin Exp Ophthalmol 245:143–147
Net M, Trias E, Navarro A, Ruiz A, Diaz P, Fontenla JR et al (2003) Cold chain monitoring during cold transportation of human corneas for transplantation. Transplant Proc 35:2036–2038
Piquot X, Delbosc B, Herve P, Montard M, Royer J (1990) Preservation of human corneas in organ culture: results of a feasibility clinical protocol. Bull Soc Ophtalmol Fr 90:429–432
Rauen U, Kerkweg U, Wusteman MC, de Groot H (2006) Cold-induced injury to porcine corneal endothelial cells and its mediation by chelatable iron: implications for corneal preservation. Cornea 25:68–77
Reim M, Althoff C, von Mulert B (1988) Effect of low temperatures on the metabolism of corneal cultures. Graefes Arch Clin Exp Ophthalmol 226:353–356
Rosenbaum K, Reinhard T, Sundmacher R (2004) Factors influencing corneal endothelium in organ cultures during transport. Ophthalmologe 101:1209–1213
Singh G, Bohnke M, Draeger J (1984) Endothelial wound healing in organ-cultured pig corneae after mechanical trauma. Fortschr Ophthalmol 81:293–295
Singh G, Bohnke M, von-Domarus D, Draeger J, Lindstrom RL, Doughman DJ (1985) Vital staining of corneal endothelium. Cornea 4:80–91
Sperling S (1978) Early morphological changes in organ cultured human corneal endothelium. Acta Ophthalmol (Copenh) 56:785–792
Sperling S, Olsen T, Ehlers N (1981) Fresh and cultured corneal grafts compared by post-operative thickness and endothelial cell density. Acta Ophthalmol (Copenh) 59:566–575
Varssano D, Russ V, Linhart Y, Lazar M (2005) Air transportation of corneal tissue: experience with local compared to transatlantic donor corneas. Cornea 24:674–677
Acknowledgements
The authors thank Dr. Estifanos Ghebremedhin for helpful statistical comments and Mrs. Christine Jaeckel for her professional technical assistance. Supported by Johann Wolfgang Goethe University (Junior Research Group Fellowship to C.M.).
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Schroeter, J., Meltendorf, C., Ohrloff, C. et al. Influence of temporary hypothermia on corneal endothelial cell density during organ culture preservation. Graefes Arch Clin Exp Ophthalmol 246, 369–372 (2008). https://doi.org/10.1007/s00417-007-0711-5
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DOI: https://doi.org/10.1007/s00417-007-0711-5