Inhibition of corneal angiogenesis by local application of vasostatin.
OBJECTIVE
This study was designed to investigate the effects of the locally supplied endogenous angiogenesis inhibitor vasostatin (VS) on corneal angiogenesis.


METHODS
Recombinant VS was expressed and purified. The effects of VS on the proliferation of endothelial cells were investigated using the methyl thiazolyl tetrazolium (MTT) assay in the absence or presence of angiogenic factors such as basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF). Corneal neovascularization was induced by implantation of hydron pellets containing bFGF in rat corneal micropockets. The potency of VS to inhibit corneal angiogenesis was investigated by incorporation of VS with bFGF in hydron pellets or topical application of VS containing eye drops to rat eyes implanted with bFGF pellets. The extent of corneal neovascularization was evaluated by microscopic and histological analyses.


RESULTS
VS potently inhibited the growth of endothelial cells in the absence or presence of angiogenic factors such as bFGF or VEGF. In the rat corneal micropocket assay, concurrent incorporation of VS abolished the bFGF induced neovascularization. When formulated in a methylcellulose eye drop, VS remained intact and functional in a 4 degrees C solution for more than 7 days. Topical application of VS eye drops potently inhibited bFGF induced neovascularization in rat corneas.


CONCLUSIONS
The present study effectively demonstrated the potential feasibility of local application of VS for treatment of corneal angiogenesis.
Wu
Pei-Chang
PC
Department of Ophthalmology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan.
Yang
Lin Cheng
LC
Kuo
Hsi-Kung
HK
Huang
Chao-Cheng
CC
Tsai
Chia-Ling
CL
Lin
Pey-Ru
PR
Wu
Ping-Ching
PC
Shin
Shyi-Jang
SJ
Tai
Ming-Hong
MH
D000287:Administration, Topical; D020533:Angiogenesis Inhibitors / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D000818:Animals; D001011:Aorta / Q000166:cytology; D037282:Calreticulin / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003001:Cloning, Molecular; D016510:Corneal Neovascularization / Q000139:chemically induced / Q000473:pathology / Q000517:prevention & control*; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000633:toxicity; D015870:Gene Expression; D008297:Male; D009883:Ophthalmic Solutions; D010446:Peptide Fragments / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000187:drug effects; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000627:therapeutic use; D020133:Reverse Transcriptase Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000633:toxicity
D020533:Angiogenesis Inhibitors; D037282:Calreticulin; D009883:Ophthalmic Solutions; D010446:Peptide Fragments; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; C121986:vasostatin; D016222:Fibroblast Growth Factor 2