Inhibition of diclofenac formulated in hyaluronan on angiogenesis in vitro and its intraocular tolerance in the rabbit eye.
OBJECTIVE
To investigate the effect of diclofenac, a potent nonsteroidal anti-inflammatory drug, formulated in hyaluronan (diclofenac/HA) on angiogenesis in vitro and its intraocular toxicity in vivo.


METHODS
The effect of diclofenac/HA on angiogenesis was determined by choriocapillary endothelial cells on Matrigel stimulated by vascular endothelial growth factor (VEGF). The tube areas were quantified by image digital analysis. For toxicity study, diclofenac/HA was injected intravitreally with a dose range from 100 to 1080 microg in 26 rabbits following gas compression vitrectomy. Potential toxicity was assessed by indirect ophthalmoscopy and by histological studies (light and electron microscopy). Retinal function was monitored by electroretinography (ERG) in six rabbits that received 400 microg of diclofenac/HA.


RESULTS
Diclofenac/HA, 180, 90 microg/ml, inhibited tube formation to 24% and 55% of the standard group (Media Ham's F12 plus 5% fetal calf serum and 50 ng/ml VEGF) respectively (P<0.01). Intravitreal injection of 540 microg or higher doses of diclofenac/HA resulted in ocular toxicity in the rabbit, demonstrated as cataract, vitreous haze and retinal damage observed by indirect ophthalmoscopy and light- and electron-microscopic examinations. No toxicity was observed in the eyes that received 400 microg or less diclofenac/HA, which was further supported by the normal ERG examined at 4 and 25 days post injection.


CONCLUSIONS
Diclofenac/HA inhibits tube formation in vitro and is non-toxic to the rabbit retina at concentrations that are inhibitory to tube formation. Our results suggest diclofenac/HA may be an effective candidate to inhibit ocular neovascularisation related to granulomatous reaction in the eye.
Shen
W Y
WY
Department of Molecular Ophthalmology, Lions Eye Institute, Perth, Western Australia, Australia. shen@cyllene.uwa.edu.au
Constable
I J
IJ
Chelva
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Rakoczy
P E
PE
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology* / Q000633:toxicity; D002196:Capillaries / Q000187:drug effects; D002478:Cells, Cultured; D002829:Choroid / Q000098:blood supply / Q000187:drug effects* / Q000473:pathology; D020256:Choroidal Neovascularization / Q000473:pathology / Q000517:prevention & control*; D004008:Diclofenac / Q000494:pharmacology* / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004359:Drug Therapy, Combination; D004596:Electroretinography; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005169:Factor VIII / Q000378:metabolism; D006820:Hyaluronic Acid / Q000494:pharmacology* / Q000633:toxicity; D008222:Lymphokines / Q000494:pharmacology; D011817:Rabbits; D012160:Retina / Q000187:drug effects* / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000894:Anti-Inflammatory Agents, Non-Steroidal; D016228:Endothelial Growth Factors; D008222:Lymphokines; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D004008:Diclofenac; D005169:Factor VIII; D006820:Hyaluronic Acid