A superfusion apparatus for ex vivo human eye irritation investigations

Elbadawy, Hossein M.; Salvalaio, Gianni; Parekh, Mohit; Ruzza, Alessandro; Baruzzo, Mattia; Cagini, Carlo; Ponzin, Diego; Ferrari, Stefano

doi:10.1016/j.tiv.2015.06.022

Cited by 7 publications

(5 citation statements)

References 19 publications

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“…The organ culture system we used is intended to reduce animal use in research (following the 3R principles). The advantages include the use of a human cornea, which is anatomically different from animal corneas, the sterile environment to prevent any interference with results, low‐cost transferable technology and research‐attractive features (Elbadawy et al ., ). This model is based on the well‐established eye irritation tests and organ culture systems using living corneas obtained from animal eyes.…”

Section: Discussionmentioning

confidence: 97%

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Effect of connexin 43 inhibition by the mimetic peptide Gap27 on corneal wound healing, inflammation and neovascularization

Elbadawy

Mirabelli

Xeroudaki

et al. 2016

British J Pharmacology

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Background and PurposeThe connexin 43 (Cx43) mimetic peptide Gap27 was designed to transiently block the function of this gap junction. This study was undertaken to investigate the effect of Gap27 on corneal healing, inflammation and neovascularization.Experimental ApproachThe effect of Gap27 on wound healing, inflammation and vascularization was assessed in primary human corneal epithelial cells (HCEC) in vitro and whole human corneas ex vivo, and in an in vivo rat wound healing model.Key ResultsGap27 enhanced the wound closure of HCEC in vitro and accelerated wound closure and stratification of epithelium in human corneas ex vivo, but did not suppress the corneal release of inflammatory mediators IL‐6 or TNF‐α in vivo. In human corneas ex vivo, F4/80 positive macrophages were observed around the wound site. In vivo, topical Gap27 treatment enhanced the speed and density of early granulocyte infiltration into rat corneas. After 7 days, the expressions of TNF‐α and TGFβ1 were elevated and correlated with inflammatory cell accumulation in the tissue. Additionally, Gap27 did not suppress VEGF release in organotypic culture, nor did it suppress early or late VEGFA expression or neovascularization in vivo.Conclusions and ImplicationsGap27 can be effective in promoting the healing of superficial epithelial wounds, but in deep stromal wounds it has the potential to promote inflammatory cell migration and accumulation in the tissue and does not suppress the subsequent neovascularization response. These results support the proposal that Gap27 acts as a healing agent in the transient, early stages of corneal epithelial wounding.

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Section: Discussionmentioning

confidence: 97%

“…The human corneal superfusion apparatus ( sa ) is an artificial human corneal environment designed and developed in our lab to mimic the human cornea in its natural environment (Elbadawy et al ., ). Organ culture corneas used for these experiments were maintained at 31°C in storage media.…”

Section: Methodsmentioning

confidence: 97%

Effect of connexin 43 inhibition by the mimetic peptide Gap27 on corneal wound healing, inflammation and neovascularization

Elbadawy

Mirabelli

Xeroudaki

et al. 2016

British J Pharmacology

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“…10,11,38 In our series, with this system, the basement membrane was irregular and also absent in some areas, whereas it is continuous and uninterrupted in other models with viable endothelial cells. Some irritancy assays use corneas mounted on an artificial anterior chamber 24,37,42 that preserves the endothelium, but these systems do not allow extended storage of corneas and are thus unsuitable for studying pathologic wound-healing processes. Tappeiner et al 43 induced an epithelial wound in porcine corneas while preserving the whole eyeball, but only for 26 hours.…”

Section: Discussionmentioning

confidence: 99%

Storage of Porcine Cornea in an Innovative Bioreactor

Guindolet

Crouzet

Hé

et al. 2017

Invest. Ophthalmol. Vis. Sci.

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“…Since the corneal epithelial cells are nourished through the ocular tear film, one of the main challenges of any ex vivo corneal model is mimicking the constant blinking that allows for uniform distribution of the precorneal tear film. Previous studies have tried to resolve this issue by either developing a mechanical means of mimicking blinking through the use of a superfusion apparatus . With the use of this machine, the investigator is able to maintain a desired temperature, the corneas exposure to media, and artificial tear flow all while in a sterile environment.…”

Section: Discussionmentioning

confidence: 99%

Development of a novel ex vivo equine corneal model

Marlo

Giuliano

Sharma

et al. 2016

Veterinary Ophthalmology

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Objective To develop an ex vivo equine corneal organ culture model. Specifically, to assess the equine cornea's extracellular matrix and cellularity after 7 days using two different culture techniques: either (a) immersion system or (b) air/liquid interface system, to determine the best ex vivo equine corneal model. Methods Fourteen healthy equine corneas with 2 mm of perilimbal sclera were freshly harvested from 7 horses undergoing humane euthanasia. One corneal-scleral ring (CSR) from each horse was randomly placed in the (a) immersion condition organ culture system (IC), with the contralateral SCR being placed in the (b) air/liquid interface organ culture system (ALC) for 7 days. All corneas were evaluated using serial daily gross photography, histology, qPCR and TUNEL assay. Results CSRs placed in the IC (a) had complete loss of corneal transparency on gross photography by 7 days, showed a significant level of corneal stromal disorganization, significantly increased αSMA levels on qPCR, and apoptosis on TUNEL assay compared to controls. The ALC (b), had weak stromal disorganization on histopathologic examination and was not significantly different from normal equine corneal controls on all other evaluated parameters. Conclusions The air-liquid interface organ culture system maintains the equine cornea's extracellular matrix and preserves corneal transparency, while the immersion condition results in near complete degradation of normal equine corneal architecture after 7 days in culture. The air-liquid organ culture is a viable option to maintain a healthy equine cornea in an ex-vivo setting for wound healing studies.

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A superfusion apparatus for ex vivo human eye irritation investigations

Cited by 7 publications

References 19 publications

Effect of connexin 43 inhibition by the mimetic peptide Gap27 on corneal wound healing, inflammation and neovascularization

Effect of connexin 43 inhibition by the mimetic peptide Gap27 on corneal wound healing, inflammation and neovascularization

Storage of Porcine Cornea in an Innovative Bioreactor

Development of a novel ex vivo equine corneal model

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