Abstract
Purpose: :
To evaluate the cytotoxic and antiproliferative properties of Bevacizumab (Avastin®) – a humanized monoclonal antibody against vascular endothelial growth factor (VEGF) – on a human retinal pigment epithelium cells line (ARPE19), a rat ganglion cell line (RGC–5), human corneal keratocytes as well as pig choroidal endothelial cells.
Methods: :
For cytotoxicity assays cells were grown to confluence and then cultured in a serum–deficient medium to ensure a static milieu. MTT assay was performed after 1 day of incubation with 0.008 mg/ml, 0.025 mg/ml, 0.08 mg/ml, 0.25 mg/ml, 0.8 mg/ml, and 2.5 mg/ml, respectively, of Bevacizumab. Nonconfluent cultures of the above mentioned cells were used to carry out proliferation assays. Bevacizumab (diluted in culture medium, concentrations as above) was added and cellular proliferative activity was monitored by BrdU–incorporation into cellular DNA after 1, 3 and 6 days.
Immune histochemistry was done by using the VEGF (C–1): sc–7269 mouse–monoclonal antibody.
Results: :
Bevacizumab was neither cytotoxic nor antiproliferative on all cell types even at the highest concentration tested. Immune histology revealed that all cultured cell types stained positive for VEGF with ARPE–19 cells being most intensively stained.
Conclusions: :
No toxic or anti–proliferative effect of bevacizumab on various ocular cell types could be observed. These results show that although the cells express VEGF, the humanised monoclonal anti–VEGF antibody has no damaging effect.
Keywords: cell survival • drug toxicity/drug effects • immunohistochemistry