May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
In-vitro Comparison of Safety Profile of AvastinTM and LucentisTM in Retinal Pigment Epithelial (ARPE-19) and Neurosensory Retinal Cells (R28)
Author Affiliations & Notes
  • A. Neekhra
    Ophthalmology, University of California-Irvine, Orange, California
  • A. L. Gramajo
    Ophthalmology, University of California-Irvine, Orange, California
  • S. Luthra
    Ophthalmology, University of California-Irvine, Orange, California
  • G. M. Seigel
    Ophthalmology, The Ross Eye Institute, University at Buffalo, SUNY, New York
  • M. C. Kenney
    Ophthalmology, University of California-Irvine, Orange, California
  • B. D. Kuppermann
    Ophthalmology, University of California-Irvine, Orange, California
  • Footnotes
    Commercial Relationships A. Neekhra, None; A.L. Gramajo, None; S. Luthra, None; G.M. Seigel, None; M.C. Kenney, None; B.D. Kuppermann, None.
  • Footnotes
    Support Discovery Eye Foundation, Iris and B. Gerald Cantor Foundation, Research to Prevent Blindness Foundation, PAAO Foundation (David & Julianna Pyott Pan-American - Retinal Research Fellowship)
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 3423. doi:
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    • Get Citation

      A. Neekhra, A. L. Gramajo, S. Luthra, G. M. Seigel, M. C. Kenney, B. D. Kuppermann; In-vitro Comparison of Safety Profile of AvastinTM and LucentisTM in Retinal Pigment Epithelial (ARPE-19) and Neurosensory Retinal Cells (R28). Invest. Ophthalmol. Vis. Sci. 2007;48(13):3423.

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

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Abstract

Purpose:: To compare the safety profile of bevacizumab (Avastin) and ranibizumab (Lucentis) on retinal cells (R28 /ARPE-19) in culture.

Methods:: ARPE-19 and R28 cells were grown to confluency and then exposed for 24 hrs to one of 3 doses of AvastinTM or LucentisTM (Genentech, San Francisco, CA). The concentrations used were 1, 2, and 5x the intravitreal clinical doses (1.25mg, 2.5mg, 6.25mg for Avastin and 0.5mg, 1.0mg, 2.5mg for Lucentis). Cellular apoptosis was quantified by measuring the mitochondrial membrane potential (ΔΨm) using the JC-1 assay. In order to determine the downstream apoptotic pathway, caspase-3 activity was measured by FLICA Apoptosis Detection kit. Cell viability studies were performed by trypan blue dye exclusion assay.

Results:: In ARPE-19 cells, ΔΨm was decreased significantly in the 5x Avastin-treated cultures compared to both Lucentis and untreated cultures (p<0.001). At 2 x, Avastin- treated cultures were lower compared to untreated cultures (p<0.05). In R28 cells, both the 5x and 2x Avastin- treated cultures had decreased ΔΨm compared to Lucentis and untreated cultures (p<0.001 and p<0.05, respectively). There was no significant difference between Avastin and Lucentis at 1x clinical dose. The caspase-3 activity was increased only in the 5x Avastin-treated cultures compared to untreated cultures (p<0.001). Cell viability was not reduced for any dose of Lucentis or Avastin compared to controls in either cell line.

Conclusions:: Both Avastin and Lucentis were non-toxic to retinal cells in culture at the clinical intravitreal doses. At 5x and 2x dose, Avastin induced early apoptotic changes as reflected by decreased mitochondrial membrane potential in both retinal cell lines. In addition, Avastin caused an increase in caspase-3 activity at 5x the clinical dose. In contrast, Lucentis was non-toxic in both retinal cell lines at all doses studied using all three assays.

Keywords: retinal neovascularization • growth factors/growth factor receptors • age-related macular degeneration 
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