Cytotoxic effects of FGF2-saporin on bovine epithelial lens cells in vitro.
OBJECTIVE
To test the ability of two preparations of FGF2-saporin, either FGF2 chemically conjugated to saporin (FGF2-SAP) or genetically engineered FGF2-saporin (rFGF2-SAP) to inhibit the growth of bovine epithelial lens (BEL) cells in vitro when in solution and when immobilized on heparin surface-modified (HSM) polymethylmethacrylate (PMMA) intraocular lenses (IOLs).


METHODS
Bovine epithelial lens cells were incubated with various concentrations FGF2-saporin for as long as 4 days. The number of surviving cells was determined by counting the number of nuclei. Because FGF2 binds to heparin, FGF2-saporin was incubated with HSM PMMA IOLs; excess toxin was washed off, and the BEL cells were grown on the FGF2-saporin-treated IOLs (HSM and non-HSM) for 4 days. Cell density was determined by image analysis.


RESULTS
Both FGF2-SAP and rFGF2-SAP were highly cytotoxic (nM range), with rFGF2-SAP 10 times less active than FGF2-SAP. FGF2-saporin bound to the surface of HSM IOLs and eluted by 2M NaCl retained its activity. Toxin bound to HSM IOLs killed more than 90% of the BEL cells placed on the IOL surface within 4 days. The ability of FGF2-saporin to prevent the growth of cells on the IOL surface was strictly dependent on the presence of heparin on the IOL.


CONCLUSIONS
FGF2-saporin is bound to HSM PMMA IOLs and prevents the growth of epithelial cells on the surface of the lens.
Behar-Cohen
F F
FF
Unité de Recherches Gérontologiques, Inserm U 118, Paris, France.
David
T
T
Buechler
Y
Y
Nova
M P
MP
Houston
L L
LL
Pouliquen
Y M
YM
Courtois
Y
Y
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000633:toxicity*; D002417:Cattle; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival; D002478:Cells, Cultured; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000633:toxicity*; D006493:Heparin; D006820:Hyaluronic Acid / Q000494:pharmacology; D000922:Immunotoxins; D007903:Lens Capsule, Crystalline / Q000187:drug effects; D007908:Lens, Crystalline / Q000166:cytology / Q000187:drug effects*; D007910:Lenses, Intraocular; D008768:Methylmethacrylates; D009699:N-Glycosyl Hydrolases; D010940:Plant Proteins / Q000633:toxicity*; D011994:Recombinant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins
D000972:Antineoplastic Agents, Phytogenic; D000922:Immunotoxins; D008768:Methylmethacrylates; D010940:Plant Proteins; D011994:Recombinant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D016222:Fibroblast Growth Factor 2; D006820:Hyaluronic Acid; D006493:Heparin; D009699:N-Glycosyl Hydrolases; D000076985:Saporins