Dehydrated form of plasmid expressing basic fibroblast growth factor-polyethylenimine complex is a novel and accurate method for gene transfer to the cornea.
OBJECTIVE
We describe a novel vector system of nonviral gene transfer into the cornea using a dehydrated form of a plasmid expressing basic fibroblast growth factor-polyethylenimine (p-bFGF-PEI) complex to induce angiogenesis.


METHODS
Corneal neovascularization was evaluated in 48 eyes of Sprague-Dawley rats after implantation of a dehydrated form of PEI containing 1 microg green fluorescent protein (p-GFP-PEI; control group), or 10 microg, 1 microg, or 0.1 microg of p-bFGF-PEI introduced by spin vacuum at ambient temperature. Neovascularization was observed and quantified from day 1 to day 45. Eighteen kDa bFGF protein expression was analyzed by Western blot and immunohistochemistry.


RESULTS
Limbal vessels began to sprout on day 3 in the p-bFGF-PEI groups. The dehydrated form of the p-bFGF-PEI complex induced dose-dependent corneal neovascularization, which reached a maximum on days 24-30 in the 10 microg bFGF group, days 18-24 in the 1 microg bFGF group, and days 15-21 in 0.1 microg bFGF group, and then regressed progressively. No neovascularization was observed in the GFP group.


CONCLUSIONS
The dehydrated form of the p-bFGF-PEI complex is a novel and precise method for controlling the dose, localizing the reagents, and avoiding loss of liquid form during transfection into corneal tissue.
Kuo
Chien-Neng
CN
Department of Ophthalmology, Chang Gung Memorial Hospital, Chiayi, Taiwan, Republic of China.
Yang
Lin-Cheng
LC
Wu
Pei-Chang
PC
Kuo
Hsi-Kung
HK
Kuo
Chian-Jue
CJ
Tai
Ming-Hong
MH
D000818:Animals; D015153:Blotting, Western; D002196:Capillaries / Q000033:anatomy & histology; D003315:Cornea / Q000098:blood supply / Q000378:metabolism*; D003890:Desiccation; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D016222:Fibroblast Growth Factor 2 / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D008297:Male; D018919:Neovascularization, Physiologic / Q000502:physiology; D010957:Plasmids; D011094:Polyethyleneimine; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors
D004337:Drug Carriers; D011993:Recombinant Fusion Proteins; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine