Abstract
Purpose :
Anti-VEGF therapy is currently an effective standard treatment for AMD; exploring the other possible pathways may provide an alternative strategy in the treatment of AMD. This includes combination of various anti-VEGF molecules such as Pazopanib, Palomid 529 and Bevacizumab (avastin) for targeting vascular endothelial growth factor (VEGF). However, the dose, time and combination effects of these molecules on VEGF-enriched choroidal vascular endothelial cells (CVECs) has not been well established in invitro. We evaluated the cytotoxicity of combination doses of Pazopanib, Palomid 529 and bevacizumab on CVECs.
Methods :
Choroidal vascular endothelial cells (CVECs) were treated with VEGF (50ng/ml) and were treated with Bevacizumab, Palomid529, Pazopanib, Bevacizumab +Palomid 529, Bevacizumab +Pazopanib, Pazopanib+Palomid 529. Cell proliferation changes analyzed with water-soluble tetrazolium salts (WST-1) assay. Cytotoxicity was evaluated by trypan blue exclusion assay at different time intervals i.e 48h, 72h, 1 week. Simultaneously reactive oxygen species levels were also measured using dihydrorhodamine 123 at similar intervals. Apoptotic marker such as caspase 3 was also measured at 72h interval.
Results :
VEGF-enriched CVECs treated with Bevacizumab, Palomid 529, Pazopanib, Bevacizumab+Palomid 529,Bevacizumab+Pazopanib, Pazopanib+Palomid 529 inhibited the proliferation of CVECs. CVECs showed a significant decrease in the cell proliferation rates when treated with combination of anti-VEGF molecules in comparison to single anti-VEGF molecules and to controls (p<0.0001). Similarly, cell viability numbers also significantly reduced in combination of anti-VEGF molecules in comparison to single anti-VEGF molecule and to controls (p<0.0001). Further, ROS levels were found to be significant in combination of anti-VEGF groups compared to drugs alone and controls (p<0.01). Similarly, apoptotic marker caspase 3 levels were found to be increased by 10 folds in (Palomid 529+ Pazopanib) group compared to any other groups and control.
Conclusions :
Although anti-VEGF molecules are effective in controlling the proliferation and cell viability rates of VEGF-enriched CVECs, a combination of these anti-VEGF molecules are more effective in inhibiting the cell proliferation rates than any one anti-VEGF molecule alone in comparison to control.
This is a 2021 ARVO Annual Meeting abstract.