Platelet-derived growth factor: receptor expression in corneas and effects on corneal cells.

PURPOSE

Platelet-derived growth factor (PDGF), a major mitogen and chemoattractant, is a dimeric molecule of disulfide-bonded A and/or B polypeptide chains (PDGF-AA/AB/BB). Two PDGF receptors (PDGFR) exist, alpha and beta, which dimerize after ligand exposure. The alpha-receptor binds both A- and B-chains, whereas the beta-receptor preferentially binds the B-chain. Whether PDGFR are present on, and whether PDGF is mitogenic for, corneal cells was investigated.

METHODS

For receptor determination, a two-step immunoperoxidase technique with monoclonal antibodies against both alpha- and beta-receptors was applied on frozen sections of human and bovine corneas. To test the mitogenic activity of PDGF-BB, two proliferation assays, the DNA synthesis assay (3H-thymidine incorporation) and the colorimetric MTT assay, were used for cultured bovine corneal endothelial cells (BCEC) and human corneal fibroblast (HCF).

RESULTS

Both receptors were present on epithelial cells, stromal fibroblasts, and endothelial cells, the beta-receptor being most abundant. In BCEC, minimal and maximal effects on DNA synthesis occurred at 10 ng/ml and 50-100 ng/ml PDGF, respectively. For HCF, the minimal and maximal effective doses were 1 ng/ml and 25-100 ng/ml of PDGF, respectively. The MTT assay, carried out in BCEC only, showed a minimal and maximal cell activity at 1 ng/ml and 10-100 ng/ml of PDGF, respectively.

CONCLUSIONS

The presence of PDGFR in human corneal epithelium, fibroblasts, and endothelium and the mitogenic effects of PDGF on corneal cells indicate that PDGF may play a role in corneal wound healing.