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1From the Department of Ophthalmology, University of Kentucky, Lexington, Kentucky; the 2Department of Ophthalmology, Medical College of Georgia, Augusta, Georgia; and the 3Department of Cell Biology and Immunology, Vrije Universiteit, Amsterdam, The Netherlands.
OBJECTIVE. To investigate the role of macrophages in the development of laser-induced choroidal neovascularization (CNV) by selective depletion with liposomal clodronate (Cl2MDP-LIP).
METHODS. Laser photocoagulation was used to induce CNV in wild-type C57BL/6J mice. Animals were treated with intravenous (IV) and/or subconjunctival (SC) Cl2MDP-LIP or PBS-LIP at the following time points: 2 days before, immediately after, 2 days before and immediately after, or 2 days after laser injury. CNV responses were compared on the basis of en masse volumetric measurements and fluorescein angiography after laser photocoagulation. Macrophages were identified by immunostaining for F4/80, and vascular endothelial growth factor (VEGF) expression was quantified by ELISA.
RESULTS. Macrophages invaded the site of laser injury within 1 day of photocoagulation and peaked at 3 days. IV Cl2MDP-LIP significantly decreased the volume of CNV and angiographic leakage when administered 2 days before and/or immediately after laser injury, but not when administered 2 days after injury. SC Cl2MDP-LIP significantly decreased lesion volume when coadministered with IV PBS-LIP but not IV Cl2MDP-LIP. IV Cl2MDP-LIP was significantly more beneficial when administered 2 days before laser injury than immediately after, but combining SC Cl2MDP-LIP with IV treatment eliminated this difference. Reduction in CNV volume correlated with VEGF protein levels and number of infiltrating macrophages.
CONCLUSIONS. Generalized macrophage depletion reduced the size and leakage of laser-induced CNV and was associated with decreased macrophage infiltration and VEGF protein. These findings define the role of the macrophage as a critical component in initiating the laser-induced CNV response.
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