Abstract
Purpose:
Chorioretinal neovasculatization (CNV) formation in animal models is essential for the investigation of the pathophysiology and treatment modalities of human neovascular chorioretinal diseases. Models are designed in order to be efficient, reproducible and repetitive over-time. The current and most popular animal model for CNV is based on disruption of the Bruch's membrane of mice retina by immobile direct-laser. This setup is inconvenient for laboratory use, difficult to conduct and expensive. Our aim was to establish a novel laser-induced model for CNV creation in pigmented mice, using a mobile indirect-diode laser and to establish the parameters of the lens, laser beam and working modality that will ensure consistent and repeatable model.
Methods:
Indirect diode laser ophthalmoscope (Iris Medical Oculight SLX System) was used. Large scale of laser power (200-800 mW) and duration (100-200 msec) was applied around the optic disc of pigmented male mice (CD57BL/6J strand), in order to achieve white bubble formation. CNV presence was searched with Hematoxylin & Eosin (H&E) and CD31 immunohistochemical staining of thin sections, and by fluorescein angiography (FA). CNV total area was determined by using CD31 staining of a flattened choroid performed at 5 and 14 days after laser. Correlations between lesion number and area of CNV were performed.
Results:
The 90D treating lens was chosen, because of the smallest depth of focus and highest accuracy in its focal point. Formation of acute vapor white bubbles was demonstrated in the laser range of 200-400 mW and 100-200 msec. CNV existence was proved at 5 and 14 days after laser by H&E and CD31 staining, and by FA. Quantitative measurements of CNV area revealed a linear correlation between CNV area and number of laser lesions performed.
Conclusions:
We described for the first time a repeatable laboratory model of CNV induction by indirect diode-laser in pigmented mice. Since indirect diode-laser is an easy-to-use, inexpensive, mobile and convenient form we suggest that it can serve as an alternative for the current immobile laser systems used for creation of CNV.
Keywords: 453 choroid: neovascularization •
578 laser •
688 retina