Basic fibroblast growth factor stimulates corneal endothelial cell growth and endothelial wound healing of human corneas.
OBJECTIVE
To determine the dose response of human recombinant basic fibroblast growth factor (bFGF) on mitogenic activity, and the supplementary role of serum in cultured bovine and human corneal endothelial cells (BCECs, HCECs). To investigate the effect of bFGF on endothelial wound healing of human corneas in vitro.


METHODS
In cell culture, DNA synthesis was assessed by 3H-thymidine incorporation. Wound healing was studied using paired human corneas after mechanical damaging of the endothelium. One cornea was treated with bFGF, and the mate served as control. Wound closure was determined after staining with trypan blue. Endothelial cell density (ECD) was assessed in the closed wound area after alizarin red staining. DNA synthesis was assessed using 3H-thymidine autoradiography.


RESULTS
In cell culture, bFGF induced a dose-dependent mitogenic response on BCECs and HCECs. Addition of serum to the culture medium shifted the dose-response curve to considerably lower bFGF concentrations. In organ culture, the time of complete wound closure shortened only marginally (0.5 day) after bFGF treatment (P < 0.01). In the closed wound center, ECD was significantly higher in 1 ng/ml bFGF-treated corneas (686 +/- 134 cells/mm2) than in controls (554 +/- 117 cells/mm2), an increase of +25%. Doses of 0.1 and 10 ng/ml also were effective, but less so than with 1 ng/ml (+11% and +15%, respectively), whereas a dose of 100 ng/ml even had a negative effect (-11%). DNA synthesis was marginally enhanced in bFGF-treated (1 ng/ml) corneas.


CONCLUSIONS
The maximal effective dose of bFGF producing a BCEC mitogenic response is dependent on serum. In human senior donor corneas, bFGF promotes endothelial wound healing predominantly by stimulation of cell migration.
Hoppenreijs
V P
VP
Department of Morphology, The Netherlands Ophthalmic Research Institute, Amsterdam.
Pels
E
E
Vrensen
G F
GF
Treffers
W F
WF
D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002417:Cattle; D002455:Cell Division; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D004247:DNA / Q000096:biosynthesis; D004261:DNA Replication; D004305:Dose-Response Relationship, Drug; D004728:Endothelium, Corneal / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D008875:Middle Aged; D008938:Mitosis; D009924:Organ Culture Techniques; D011994:Recombinant Proteins / Q000494:pharmacology; D014945:Wound Healing
D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D004247:DNA