Endothelial cell density in porcine corneas after exposure to hypotonic solutions

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

doi:10.1007/s00417-006-0334-2

Cited by 6 publications

(4 citation statements)

References 16 publications

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“…3A). The variability between tested tissue was minimal and these results prove similar to reported porcine endothelial densities, ranging from 2500-4000 cells/mm 2 and corneal thickness, ranging from 0.8 to 1.2 mm [19][20][21][22][23] . For assessing tissue storage, we assessed eyes 1 or 7 days after storage at 4 °C (submerged in HBSS), −20 °C (wrapped in HBSS soaked gauze), and −80 °C (wrapped in HBSS soaked gauze) ( Table 1 for experimental group layout).…”

Section: Resultssupporting

confidence: 83%

Development and Characterization of a Benchtop Corneal Puncture Injury Model

Snider

Cornell

Acevedo

et al. 2020

Sci Rep

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During recent military operations, eye-related injuries have risen in frequency due to increased use of explosive weaponry which often result in corneal puncture injuries. these have one of the poorest visual outcomes for wounded soldiers, often resulting in blindness due to the large variations in injury shape, size, and severity. As a result, improved therapeutics are needed which can stabilize the injury site and promote wound healing. Unfortunately, current corneal puncture injury models are not capable of producing irregularly shaped, large, high-speed injuries as seen on the battlefield, making relevant therapeutic development challenging. Here, we present a benchtop corneal puncture injury model for use with enucleated eyes that utilizes a high-speed solenoid device suitable for creating militaryrelevant injuries. We first established system baselines and ocular performance metrics, standardizing the different aspects of the benchtop model to ensure consistent results and properly account for tissue variability. The benchtop model was evaluated with corneal puncture injury objects up to 4.2 mm in diameter which generated intraocular pressure levels exceeding 1500 mmHg. Overall, the created benchtop model provides an initial platform for better characterizing corneal puncture injuries as seen in a military relevant clinical setting and a realistic approach for assessing potential therapeutics. Eye-related injuries have risen in recent combat operations due to increased use of explosive weaponry. From World War I to Operations Iraqi and Enduring Freedom, eye-related injuries increased from 2% to 13% of combat injuries, respectively 1. The most prevalent ocular injuries are corneal abrasions, lacerations, and full thickness punctures, resulting in vision impairment without timely intervention 2. While there are treatments readily available for superficial corneal damage, more severe injuries such as full-thickness corneal punctures (CP) are challenging to treat. CP wounds predominantly enter through the cornea and may damage the iris, lens, and even retina, depending on the speed, material, and trajectory of how the eye was punctured 3. These injuries are not mitigated until injured warfighters are evacuated back to an ophthalmic specialist for surgical repair. For CP injuries where intraocular foreign bodies are present, the average time between point of injury and surgical repair was 21 days in recent combat operations 4. An open, untreated CP injury is highly susceptible to infection, metallic poisoning, and reduced intraocular pressure (IOP). Further, low IOP results in poor nutrient delivery throughout the eye, impacting viability and function of ocular tissues 3,5,6. As a result, CP injuries result in poor visual outcomes even after medical intervention 7. To date, when CP injuries occur on the battlefield the chain of treatment first involves placing a rigid eye shield on the injury site until evaluation by an ophthalmic specialist. If no intraocular foreign body (IOFB) is present, sutures are the gold...

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Section: Resultssupporting

confidence: 83%

Development and Characterization of a Benchtop Corneal Puncture Injury Model

Snider

Cornell

Acevedo

et al. 2020

Sci Rep

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“…To demonstrate the loss of irreversibly damaged endothelial cells, corneal discs were therefore not assessed directly after exposure but after further culturing for 48 h as previous published [16][17][18]. Removal of peripheral areas of the cornea, which show higher cell density than the cornea's center, was done to prevent repair of endothelial damage by means of spreading of peripheral endothelial cells.…”

Section: Discussionmentioning

confidence: 99%

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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“…To demonstrate the loss of irreversibly damaged endothelial cells, corneal discs were therefore not assessed directly after exposure but after culturing the corneas for 48 hours as previously published [2,5,12]. Removal of peripheral areas of the cornea, which show higher cell density than the cornea's center, was done to prevent repair of endothelial damage by means of spreading of peripheral endothelial cells.…”

Section: Discussionmentioning

confidence: 99%

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

Schroeter

Meltendorf

Ohrloff

et al. 2007

Graefes Arch Clin Exp Ophthalmol

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Exposure of organ-cultured porcine corneas in dextran containing medium to 4 degrees C for 12 hours and 21 degrees 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 degrees 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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Endothelial cell density in porcine corneas after exposure to hypotonic solutions

Abstract: Exposure to 1.8% sucrose for 4 min induces a significant endothelial cell loss of 10% on average, whereas exposure to hypotonic BSS did not significantly influence the endothelial cell density.

Cited by 6 publications

References 16 publications

Development and Characterization of a Benchtop Corneal Puncture Injury Model

Development and Characterization of a Benchtop Corneal Puncture Injury Model

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

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

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