September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Influence of different lutein-based dyes on corneal endothelial cell viability
Author Affiliations & Notes
  • Anja Katharina Gruenert
    Department of Ophthalmology, University Erlangen Nurnberg, Erlangen, Germany
  • Sara Sousa
    Kemin Pharma, Barcarena, Portugal
  • Marta Czugala
    Department of Ophthalmology, University Erlangen Nurnberg, Erlangen, Germany
  • Friedrich E Kruse
    Department of Ophthalmology, University Erlangen Nurnberg, Erlangen, Germany
  • Gerd Geerling
    Departmentment of Ophthalmology, University Dusseldorf, Dusseldorf, Germany
  • Thomas Armin Fuchsluger
    Department of Ophthalmology, University Erlangen Nurnberg, Erlangen, Germany
  • Footnotes
    Commercial Relationships   Anja Gruenert, Kemin Pharma (F); Sara Sousa, Kemin Pharma (E); Marta Czugala, None; Friedrich Kruse, None; Gerd Geerling, None; Thomas Fuchsluger, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 5265. doi:
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      Anja Katharina Gruenert, Sara Sousa, Marta Czugala, Friedrich E Kruse, Gerd Geerling, Thomas Armin Fuchsluger; Influence of different lutein-based dyes on corneal endothelial cell viability. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5265.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : In intraocular surgery different dyes can be used for a better visualization of membranes. Applications in the anterior segment of the eye include capsule staining during cataract surgery and staining of the endothelium-Descemet membrane layer during Descemet membrane endothelial keratoplasty (DMEK). Although cytotoxic effects on corneal endothelial cells (CEC) have been described, trypan blue is the most commonly used dye. The aim of this study is to compare alternative lutein-based dyes to dyes containing trypan blue without lutein in terms of their effect on CEC viability.

Methods : CEC and human donor corneas were incubated with different commercially available dyes (VisionBlue®, PhacodyneTM and RetidyneTM Cap) for different time periods (1, 5, 15, 30, 60, 120 and 240 minutes). Viability of CEC was assessed by flow cytometry as well as by confocal microscopy. Furthermore, metabolic activity was analyzed using Cell Counting Kit-8 (CCK-8) assay.

Results : After incubation of CEC with VisionBlue®, PhacodyneTM and RetidyneTM Cap for up to 4 hours mean percentage of apoptotic and of dead cells did not differ significantly from negative control (one-way ANOVA test, p=0.185). The percentage of apoptotic and of dead cells increased with increasing incubation time. The amount of apoptotic cells ranged from 0.8% to 5.0% with VisionBlue®, from 0.7% to 5.6% with PhacodyneTM, and from 1.0% to 5.8% with RetidyneTM Cap. Mean percentage of dead cells was below 2.5% with all tested dyes and all incubation periods. However, after incubation of CEC with VisionBlue® for longer than 60 minutes significant cell loss due to detachment of cultured cells was observed. In human donor corneas incubation with each of the dyes for a period of 120 and 240 minutes resulted in significant loss of endothelial cells compared to negative control. Metabolic activity of CEC decreased with increasing incubation time. Cells incubated with VisionBlue® showed a significantly lower metabolic activity compared to those incubated with PhacodyneTM and RetidyneTM Cap (e.g. after 60 minutes of incubation: 0.63 versus 0.85 and 0.83 relative to negative control).

Conclusions : The lutein-based dyes PhacodyneTM and RetidyneTM Cap seem to have less cytotoxic effects on corneal endothelium compared to VisionBlue®. These data suggest that the use of lutein-based intraocular dyes is more advantageous than trypan blue-based dyes without lutein.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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