Purchase this article with an account.
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.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
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.
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.
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).
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.
This PDF is available to Subscribers Only