Abstract
Purpose: :
Bevacizumab (Avastin®, Genentech, California, USA), is an anti–VEGF recombinant humanized monoclonal IgG1 antibody used to treat colorectal cancers. Bevacizumab may have a role in treating ocular disorders involving fibrovascular proliferation. This study explores morphologic effects of bevacizumab on corneal stromal fibroblasts in vitro.
Methods: :
Human corneal fibroblasts were propagated from explanted human donor cornea tissue. Once monolayers had reached confluence, the fibroblasts were passaged and re–cultured. The fibroblasts were seeded at a concentration of 10,000 fibroblasts per culture well and incubated overnight with growth media. Monolayers were then washed and covered with two bevacizumab concentrations: 1.25mg/0.05ml and 0.6 mg/0.05 ml. This process was repeated with two control solutions. The first control solution contained the same bevacizumab buffer solution with IgG replacing the anti–VEGF antibody. The second control contained buffer solution only. The monolayers were examined and photographed at 10 and 30 minutes post treatment and were then washed, reconstituted in growth media, incubated overnight and evaluated for recovery. Fibroblast morphology was assessed using phase contrast microscopy per x10 and x20 field.
Results: :
Treated corneal stromal fibroblasts exhibited significant morphological changes compared with control fibroblasts. Changes noted included lost cell–to–cell interaction and adhesion properties as compared to controls. The morphological changes were dose–dependent with the higher bevacizumab concentration showing more pronounced alteration. Bevacizumab treated fibroblasts did not recover as well in growth media as indicated by the lack of cell proliferation 24 hours later.
Conclusions: :
Bevacizumab appears to decrease corneal stromal fibroblasts survival in–vitro. This may represent a new therapeutic modality for treating corneal fibrovascular diseases. More studies are needed to evaluate the effect of this medication prior to clinical use.
Keywords: cornea: stroma and keratocytes • cell adhesions/cell junctions • neovascularization