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L.D. Waterbury; Amfenac, Diclofenac and Ketorolac Lack Anti–Angiogenic Activity in Cultured HUVEC Cells . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1753.
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To evaluate the anti–angiogenic activity of ophthalmic NSAIDs (amfenac, diclofenac and ketorolac) in cultured HUVEC cells.
HUVEC cells obtained from American Type Culture Collection–CRL–1730 were cultured in 96 well plates at 37º C and incubated for 18 hours in the presence and absence of test agent. Amfenac, diclofenac, and ketorolac were tested at 0.1 nM, 1.0 nM, 10 nM, 0.1 µM, and 1.0 µM. Suramin (10 µM) was used as a reference standard. At the end of the incubation, the morphology of the endothelial cell tubes in the individual wells was evaluated by photomicroscopy. Cells were photographed and total tube length was determined. The failure to form continuous networks between cell bodies and tube disruption was used as an index of anti–angiogenic activity.
None of the NSAIDs inhibited tube formation for more than 30% even when tested at the highest dose (1.0 µM). Amfenac, diclofenac and ketorolac were inactive even at the highest dose tested of 1.0 µM. The reference standard suramin resulted in a complete inhibition of angiogenesis at 30 µM. The maximum concentration tested of 1.0 µM exceeds that reported for any of the aforementioned drugs in the back of the eye. Using HUVEC cells, amfenac present at a 10–fold higher concentration of 1.0 µM lacked anti–angiogenic activity.
Ophthalmic NSAIDs are not anti–angiogensis agents as shown by this comparison using HUVEC. Only suramin (the reference standard) displayed significant anti–angiogenic activity.
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