Influence of temporary hypothermia on corneal endothelial cell density during organ culture preservation

Schroeter, Jan; Meltendorf, Christian; Ohrloff, C.; Rieck, Peter

doi:10.1007/s00417-007-0711-5

Cited by 15 publications

(7 citation statements)

References 16 publications

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“…It was interesting to find a slight reduction of cell density despite an evident endothelial cell mortality and low pleomorphism rate of endothelial mosaic, suggesting a delayed re-arrangement of the endothelium layer secondary to the reduced rate of cell physiology because of the low storage temperature 18. Corneal thickness of posterior lenticules increased by approximately 20% after 24 h of cold storage in both groups examined.…”

Section: Discussionmentioning

confidence: 95%

Pre-cut donor tissue for Descemet stripping automated keratoplasty: anterior hinged lamella on versus off

Amato¹,

Oddone

Nubile

et al. 2009

British Journal of Ophthalmology

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Section: Discussionmentioning

confidence: 95%

Pre-cut donor tissue for Descemet stripping automated keratoplasty: anterior hinged lamella on versus off

Amato¹,

Oddone

Nubile

et al. 2009

British Journal of Ophthalmology

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“…In order to prevent corneal damage procedural guidelines for cornea banks recommend that transportation of organ cultured corneoscleral buttons should be performed at temperatures between 10 and 40°C [7]. This recommendation is consistent with our previous findings that storage of corneal grafts at temperatures down to 4°C for 12 h does not seem to damage the endothelial cell layer [8].…”

Section: Introductionmentioning

confidence: 50%

Impact of temporary hyperthermia on corneal endothelial cell survival during organ culture preservation

Schroeter¹,

Ruggeri

Thieme

et al. 2015

Graefes Arch Clin Exp Ophthalmol

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To evaluate temporary exposure to hyperthermia for its impact on endothelial cell density of porcine corneas in organ culture medium containing dextran with regards to possible negative influences of high temperatures during the storage and transport of corneal grafts. Four groups of central discs (diameter 8 mm) from the corneas of both eyes in 40 pigs were first organ-cultured (MEM with 6% dextran 500) for 24 h at 32°C. Ten corneas were then exposed to 40°C in group 1, to 42°C in group 2, to 44°C in group 3, and to 50°C in group 4 for 12 h each. The paired corneal discs for all groups were not treated, stored at 32°C and served as controls. After further organ culture of all corneas for 48 h 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 on three central images using a system for the automatic estimation of morphometric parameters of corneal endothelium. Exposure for 12 h to 40°C as well as to 42°C induced no endothelial cell loss. Statistical analysis showed no significant difference of the endothelial cell density between corneas exposed to 40°C and 42°C and the control corneas (40°C treatment: 4736 ± 426 cells/mm(2) and control: 4762 ± 344 cells/mm(2), p = 0.74; 42°C treatment: 4240 ± 363 cells/mm(2) and control: 4176 ± 448 cells/mm(2), p = 0.40). Exposure to 44°C and 50°C lead to total necrosis of the endothelial cell layer. Exposure of organ cultured porcine corneas in dextran containing medium up to 42°C for 12 h does not compromise the endothelial cell density in a clinically relevant manner. Temperatures above 42°C, as it might be the case during transports from the cornea bank to the ophthalmic surgeon, must be strictly avoided as they damage the endothelial cell layer.

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“…There has been considerable research examining the whole cornea and isolated endothelial cells in relation to their biology, preservation and transplantation, including investigations into the role that molecular alterations have in different corneal models[3,5,11,13,27,39-41,44,46,49,54,56]. Some studies have focused on the role that storage temperature has on endothelial survival, as storage at normothermic temperatures (organ culture) can have some benefits over hypothermic storage [38,41-43,47,49].…”

Section: Introductionmentioning

confidence: 99%

The unfolded protein response in human corneal endothelial cells following hypothermic storage: Implications of a novel stress pathway

2011

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Human corneal endothelial cells (HCEC) have become increasingly important for a range of eye disease treatment therapies. Accordingly, a more detailed understanding of the processing and preservation associated stresses experienced by corneal cells might contribute to improved therapeutic outcomes. To this end, the unfolded protein response (UPR) pathway was investigated as a potential mediator of corneal cell death in response to hypothermic storage. Once preservation-induced failure had begun in HCECs stored at 4°C, it was noted that necrosis accounted for the majority of cell death but with significant apoptotic involvement, peaking at several hours post-storage (4-8 hours). Western blot analysis demonstrated changes associated with apoptotic activation (caspase 9, caspase 3, and PARP cleavage). Further, the activation of the UPR pathway was observed through increased and sustained levels of ER folding and chaperone proteins (Bip, PDI, and ERO1-Lα) in samples experiencing significant cell death. Modulation of the UPR pathway using the specific inhibitor, salubrinal, resulted in a 2-fold increase in cell survival in samples experiencing profound cold-induced failure. Furthermore, this increased cell survival was associated with increased membrane integrity, cell attachment, and decreased necrotic cell death populations. Conversely, addition of the UPR inducer, tunicamycin, during cold exposure resulted in a significant decrease in HCEC survival during the recovery period. These data implicate for the first time that this novel cell stress pathway may be activated in HCEC as a result of the complex stresses associated with hypothermic exposure. The data suggest that the targeted control of the UPR pathway during both processing and preservation protocols may improve cell survival and function of HCEC thus improving the clinical utility of these cells as well as whole human corneas.

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Influence of temporary hypothermia on corneal endothelial cell density during organ culture preservation

Cited by 15 publications

References 16 publications

Pre-cut donor tissue for Descemet stripping automated keratoplasty: anterior hinged lamella on versus off

Pre-cut donor tissue for Descemet stripping automated keratoplasty: anterior hinged lamella on versus off

Impact of temporary hyperthermia on corneal endothelial cell survival during organ culture preservation

The unfolded protein response in human corneal endothelial cells following hypothermic storage: Implications of a novel stress pathway

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