June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Evaluation of Cytotoxicity of Anti-VEGF Molecules on Vascular Endothelial Growth Factor Enriched Choroidal Vascular Endothelial Cells
Author Affiliations & Notes
  • Bharani Krishna Mynampati Arunadithya
    Ophthalmology, University of Florida Health, Jacksonville, Florida, United States
  • Sandeep grover
    Ophthalmology, University of Florida Health, Jacksonville, Florida, United States
  • Footnotes
    Commercial Relationships   Bharani Krishna Mynampati Arunadithya, None; Sandeep grover, None
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 231. doi:
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      Bharani Krishna Mynampati Arunadithya, Sandeep grover; Evaluation of Cytotoxicity of Anti-VEGF Molecules on Vascular Endothelial Growth Factor Enriched Choroidal Vascular Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2021;62(8):231.

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      © ARVO (1962-2015); The Authors (2016-present)

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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.

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