April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Effects of Different Intravitreal Treatments on the Phagocytosis of Retinal Dying Cells and Inflammation
Author Affiliations & Notes
  • G. Petrovski
    Department of Ophthalmology,
    University of Debrecen, Debrecen, Hungary
  • E. Berenyi
    Department of Biochemistry and Molecular Biology,
    University of Debrecen, Debrecen, Hungary
  • M. C. Moe
    Department of Ophthalmology, Centre for Eye Research, University of Oslo, Oslo, Norway
  • A. Vajas
    Department of Ophthalmology,
    University of Debrecen, Debrecen, Hungary
  • L. Fesus
    Department of Biochemistry and Molecular Biology,
    University of Debrecen, Debrecen, Hungary
  • A. Facsko
    Department of Ophthalmology,
    University of Debrecen, Debrecen, Hungary
  • A. Berta
    Department of Ophthalmology,
    University of Debrecen, Debrecen, Hungary
  • Footnotes
    Commercial Relationships  G. Petrovski, None; E. Berenyi, None; M.C. Moe, None; A. Vajas, None; L. Fesus, None; A. Facsko, None; A. Berta, None.
  • Footnotes
    Support  Mecenatura Grant, University of Debrecen, Hungary
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 3698. doi:
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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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Abstract

Purpose: : To study the effect of different intravitreal treatments on the phagocytosis of dying retinal pigment epithelial cells and the inflammatory response associated with it.

Methods: : 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.

Results: : 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.

Conclusions: : 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.

Keywords: retina • phagocytosis and killing • inflammation 
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