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G. Petrovski, E. Berenyi, M. C. Moe, A. Vajas, L. Fesus, A. Facsko, A. Berta; Effects of Different Intravitreal Treatments on the Phagocytosis of Retinal Dying Cells and Inflammation. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3698.
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To study the effect of different intravitreal treatments on the phagocytosis of dying retinal pigment epithelial cells and the inflammatory response associated with it.
Two different death patterns were induced in vitro in ARPE-19 cells: death through detachment from the extracellular matrix on polyHEMA coated surfaces known as anoikis and UV-induced apoptosis. Two-colored phagocytic assays were carried out where the phagocytes (human monocyte-derived macrophages) engulfed the dying cells under different treatment modalities (triamcinolone (1uM), bevacizumab (312.5ug/mL), pegaptanib (75ug/mL) and ranibizumab (125ug/mL)). Flow cytometric analysis (FACS Calibur) was used to quantify the phagocytic process as well as measure the released amounts of IL-1β, IL-6, IL-8, IL-10 and TNFalpha using a Cytokine Bead Array.
Macrophages engulfed the dying anoikic and apoptotic ARPE-19 cells at a similar and increasing rate over 8 hours of co-incubation (11.2+/-1.7% and 10.5+/-1.2% at 8 hours, respectively). Their phagocytic capacity increased by 2.1+/-0.3 times during engulfment of both types of dying cells under triamcinolone treatment, but remained unchanged for all other treatments. In the case of UV-induced dying ARPE-19 cells, probably due to presence of secondary necrosis, increased IL-6 and IL-1beta levels were detected which could be suppressed by triamcinolone, but not the other three treatments.
The macrophage mediated clearance of different dying ARPE-19 cells can serve as a good in vitro model for studying wet AMD and for testing different pharmacological and inflammatory aspects of this process.
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