April 2011
Volume 52, Issue 14
ARVO Annual Meeting Abstract  |   April 2011
Effect of Solubilized and Crystalline Loteprednol Etabonate in ARPE-19 and R28 Cells In Vitro
Author Affiliations & Notes
  • Vishal R. Sharma
    Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • A L. Gramajo
    Ophthalmology, Centro de Ojos Romagosa-Fundacion VER, Cordoba, Argentina
  • G M. Seigel
    Center for Hearing and Deafness, SUNY at Buffalo, Buffalo, New York
  • M C. Kenney
    Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • B D. Kuppermann
    Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • Footnotes
    Commercial Relationships  Vishal R. Sharma, None; A. L. Gramajo, None; G. M. Seigel, None; M. C. Kenney, None; B. D. Kuppermann, None
  • Footnotes
    Support  Discovery Eye Foundation, Guenther Foundation, Lincy Foundation, Iris and B. Gerald Cantor Foundation, Polly and Michael Smith Foundation, Research to Prevent Blindness.
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 5636. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Vishal R. Sharma, A L. Gramajo, G M. Seigel, M C. Kenney, B D. Kuppermann; Effect of Solubilized and Crystalline Loteprednol Etabonate in ARPE-19 and R28 Cells In Vitro. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5636.

      Download citation file:

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

  • Supplements

Purpose: : To study the effects of Loteprednol Etabonate 0.5% suspension (LE, Lotemax®, Bausch&Lomb, Tampa, FL), its solubilized form, and vehicle on human retinal pigment epithelial cells (ARPE-19) and rat neurosensory retinal cells (R28) in vitro.

Methods: : ARPE-19 and R28 cells were treated for 2, 6, or 24 hours with LE or DMSO solubilized LE (S-LE) at 250 µg/ml (2X clinical dose), 125 µg/ml (1X ), 62.5 µg/ml (X/2), 31.25 µg/ml (X/4), and 15.62 µg/ml (X/8). Cell viability (CV) was measured using the trypan blue dye-exclusion assay or Calcein Assay. Equivalent concentrations of DMSO were used as controls. The vehicle was obtained as the supernatant after centrifugation of LE. Lactose dehydrogenase (LDH), a measure of necrosis, was quantified using the LDH-Cytotoxicity Assay.

Results: : In both cell lines LE at 24 hours caused a significant (p<0.01) decrease in CV at all concentrations except at the lowest dose (X/8 clinical dose) in ARPE-19 cells. ARPE-19 cell CV at 24 hrs at LE doses of X, X/2, X/4 and X/8 were 8.2±3.8%, 80.4±0.9%, 80.5±1.5% and 94.4±2.3%. R28 cell 24 hour CV were 22.3±3.1%, 58.8±4.6%, 69.7±8.6% & and 76.1±6.5%. Control CV was normalized to 100 %. S-LE at 24 hours was found to be toxic to ARPE-19 and R 28 cells only at 2X dose (59.6±7.8% for ARPE-19 versus 88.1±4.4% for controls (p=0.01) and 52.3±2.9% for R28 versus 64.9±2.5 % for controls [p=0.03]). LDH was significantly elevated for ARPE-19 cells and R 28 cells treated with 2X S-LE (0.55±0.02 for ARPE-19 versus 0.37±0.01 for control, (p<0.001); 0.14 ± 0.001 for R28 versus 0.13 ± 0.003 for control [p<0.001]). Vehicle decreased CV at 2X and X in ARPE-19 cells (12.6 ± 2.1 and 89.4±3.5 for 2X and X versus 98.2±0.5 for controls (p<0.0001 and p= 0.05 respectively). Vehicle reduced CV of R28 cells only at 2X (53.2±3.8 versus 98.2±0.5 for controls; p<0.001).

Conclusions: : Loteprednol Etabonate 0.5% suspension was cytotoxic at all doses to both cell lines, except X/8 clinical dose for ARPE-19. Solubilized LE and vehicle were toxic at 2X clinical dose. Based on LDH results, cytotoxicity from LE maybe due to necrosis.

Keywords: corticosteroids • retinal pigment epithelium • inflammation 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.