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Felix Bock, Gabór Tóth, Nora Szentmary, Franziska Bucher, Claus Cursiefen; Corneal Crosslinking with Ultraviolet-A and Riboflavin in a murine model of corneal neovascularisation. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4517.
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© ARVO (1962-2015); The Authors (2016-present)
To analyse the impact of corneal crosslinking, using riboflavin and UVA-light illumination on existing blood and lymphatic vessels in the cornea.
We used the established murine model of suture-induced inflammatory corneal neovascularisation assay. 14 days after suture placement we removed the sutures.<br /> The mice underwent corneal abrasion and received cross-linking treatment using 0.1% riboflavin-5-phosphat solution (Mediocross H) (1x riboflavin-5-phosphat drop in 15 minutes) and subsequent UVA-light-illumination (370 nm; irradiance, 3 mW/cm2; dose, 5.4 J/cm2, 1x riboflavin-5-phosphat drop) for 15 minutes.<br /> After four days, corneal blood and lymphatic vessels were stained ex vivo using LYVE-1 and CD31 antibodies. Depth of cross linking was assessed by measuring the nucleus free area between Bowmann's layer and corneal endothelium using Dapi staining and an ApoTome2 microscope.
The cross linked corneas showed an enhanced rigidity as a sign of successful cross linking and were not perforated. The corneal stroma was cross linked to a mean depth of 23.96% (SD 18.5%) of total corneal stromal thickness. Furthermore we found a significant negative correlation between the depth of cross linking and the area covered by blood vessels (p=0.018; r2= 0.881). Epithelial healing was delayed in the crosslinked animals.
We demonstrate a novel murine model of corneal crosslinkink. Crosslinking in murine corneas is a new way to analyse the effect of this clinically often used method on corneal cells. This pilot study shows that crosslinking can regress pathological corneal blood vessel in a depth dependent manner. Therefore this is a promising model to develop a new strategy to regress preexisting corneal vessels prior to keratoplasty.
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