1. EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION Selim Doganay, MD¹; Penpe Gul Firat, MD¹; Cem Cankaya, MD¹; Hale Kirimlioglu, MD 2 Inonu University School of Medicine, Malatya-Turkey ¹ Department of Ophthalmology, ² Department of Pathology, Authors have no financial or proprietary interest in any instrument or products used in this study

2. Purpose  To evaluate effects of resveratrol and bevacizumab on experimental corneal neovascularization. EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

3. Introduction  It has been shown that vascular endothelial growth factor (VEGF) stimulates corneal neovascularization in animal models. 1-4  Bevacizumab is a recombinant humanized murine monoclonal antibody that binds, and inhibits the biological activity of all human VEGF-A isoforms. 5-7  Resveratrol is an agent which directly inhibits the tumor growing in an unknown mechanism. It has been shown that resveratrol has anti-angiogenic properties. 8,9

4. Materials and methods  Corneal alkali burn was performed by touching central cornea with a standard 7 mm diameter 1 N NaOH-soaked Whatman 3 filter paper for 2 minutes in 62 eyes of 31 male white Vienna rabbits.  We waited seven days for the effect of chemical cauterization to be appear. The rabbits were then divided randomly into four groups. EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

5. (n: Number of eyes, DMS: Dimetil sulfoksid, NaCl: Sodium clor solution n drug Group 116 Resveratrol ( 10 mg/ml- 0.05 ml, solved in DMS solution) Group 216 DMS (0.05 ml) Group 316 Bevacizumab ( 10 mg/ ml 0.05 ml, diluated in % 0.9 NaCl) Group 414 NaCl (0.05 ml % 0.9) Drugs were administered to the both eyes of rabits by subconjunctival injection for 7 days EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

6.  Corneal photos were taken by Nikon FS-3 biomicroscopy 15 days after alkali injury.  Inflamatuar index (ciliary injection, central corneal edema, peripheral corneal edema) was analyzed as previously described. 10  Corneal neovascularization areas were calculated as unit square on photograps with Auto CAD- 2005 program in all groups. EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

7. Results  According to inflamatuar index scores, there was no significant difference among the group 1-2 and group 2-4 (p<0.008- Bonferoni Mann- Whitney U test)  There was a significant difference according to the inflamatuar index scores among the group 2-3 and group 3-4. EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

8.  A significant difference was obtained between the group 1 and group 3 for the central and peripheral corneal edema scores.  The calculation of corneal neovascularization area in group 3 was significantly smaller than the other groups. EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

9. Bevacizumab DMSO NACL Resveratrol EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

10. 709.43 713.61 50.28 711.83 0 100 200 300 400 500 600 700 800 1234 Groups New Vascularization area (Unit square) EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

11. Conclusions  Subconjunctival administration of bevacizumab inhibits corneal neovascularization effectively in the rabbit corneal alkali burn model.  No effect of resveratrol to the corneal neovascularization on experimental model of corneal alkali burn was seen at the doses of usage. EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION

12. References 1.Gan L, Fagerholm P, Palmblad J. Vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 in the regulation of corneal neovascularization and wound healing. Acta Ophthalmol Scand 2004;82:557-563. 2.Chang CH, Gabison EE, Kato D, Azar DT. Corneal neovascularization. Curr Opin Ophthalmol 2001;12:242-249. 3.Epstein RJ, Stulting RD, Hendricks RL, Harris DM. Corneal neovascularization. Pathogenesis and inhibition. Cornea 1987;6:250-257. 4.Gan L, Fagerholm P, Palmblad J. Vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 in the regulation of corneal neovascularization and wound healing. Acta Ophthalmol Scand 2004;82:557-563. 5.Spaide RF, Laud K, Fine HF, et al. Intravitreal bevacizumab treatment of chroidal neovascularization secondary to age-related macular degeneration. Retina 2006;26:383-90. 6.Lynch SS, Cheng CM. Bevacizumab for neovascular ocular diseases. Ann Pharmacother 2007;41:614-25. 7.Kahook MY, Schuman JS, Noecker RJ. Intravitreal bevacizumab in apatient with neovascular glaucoma. Ophthalmic Surg Lasers Imaging 2006;37:144-6. 8.Dulak J. Nutracetical as anti-angiogenic agents:hopes and reality. J Phiysiol Pharmacol 2005;56:51-69. 9.Doganay S, Borazan M, Iraz M, Cigremis Y. The effect of resveratrol in experimental cataract model formed by sodium selenite. Curr Eye Res 2006;31:147-53. 10.Wu PC, Liu CC, Chen CH, et al. Inhibition of experimental angiogenesis of cornea by somatostatin. Graefes Arch Clin Exp Ophthalmol 2003 ;241:63-9. EFFECT OF RESVERATROL AND BEVACIZUMAB ON EXPERIMENTAL CORNEAL NEOVASCULARIZATION