Abstract
Purpose:
To demonstrate that minocycline protects ARPE-19 cells from the effects of hypoxia in cell culture
Methods:
ARPE-19 cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum, penicillin and streptomycin and seeded onto 6 well plates at a concentration of 5 X 104 cells per well. Minocycline concentrations between 2μM and 10μM were added to the culture media. Cell growth curves with and without minocycline were constructed for cells cultured in an hypoxic chamber (2% oxygen) and results compared to culture in normoxic conditions over a period of 8 days. (n=8 for each growth curve) A Scepter® handheld cell counter was used to measure cell counts which were conducted in triplicate.
Results:
A Mann Whitney U test showed that there was a significant difference in ARPE-19 growth between normoxic and hypoxic conditions when the culture media was supplemented with Minocycline at concentrations between 2μM and 10μM at all points of the cell growth curve. (p < 0.04)
Conclusions:
Cell and molecular biological analyses suggest that chronic oxidative stress and inflammation are involved in the pathogenesis of AMD. Hypoxia may result from increased metabolic demand during inflammation or poor perfusion in the central macula due to vessel stenosis. This study demonstrates that minocycline protects retinal pigment epithelial cells in culture from the effects of hypoxia. This suggests minocycline may have a therapeutic role in the treatment of age related macular degeneration.
Keywords: 412 age-related macular degeneration •
426 apoptosis/cell death •
694 retinal culture