June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
ASCORBIC ACID ATTENUATES HYDROGEN PEROXIDE INDUCED OXIDATIVE STRESS AND OSTEOBLASTS DEMONSTRATE ANTIOXIDANT RECYCLING POTENTIAL
Author Affiliations & Notes
  • Richard Tran
    University of Colorado Denver School of Medicine, Aurora, Colorado, United States
  • Joshua Morgenstern
    University of Colorado Denver School of Medicine, Aurora, Colorado, United States
  • Steven Droho
    University of Colorado Denver School of Medicine, Aurora, Colorado, United States
  • Jeffrey L Olson
    University of Colorado Denver School of Medicine, Aurora, Colorado, United States
  • Footnotes
    Commercial Relationships   Richard Tran None; Joshua Morgenstern None; Steven Droho None; Jeffrey Olson None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 375 – F0206. doi:
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      Richard Tran, Joshua Morgenstern, Steven Droho, Jeffrey L Olson; ASCORBIC ACID ATTENUATES HYDROGEN PEROXIDE INDUCED OXIDATIVE STRESS AND OSTEOBLASTS DEMONSTRATE ANTIOXIDANT RECYCLING POTENTIAL. Invest. Ophthalmol. Vis. Sci. 2022;63(7):375 – F0206.

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

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Abstract

Purpose : Oxidative stress is strongly implicated in disease progression of age-related macular degeneration. Oral supplements, including ascorbic acid (AA), target this oxidative etiology, yet efficacy is limited due to insufficient ocular distribution. One possible avenue is restoring the antioxidant potential of the vitreous by improving recycling of the inactive oxidized form of AA, dehydroascorbic acid (DHA), back to its active reduced form. Here, we demonstrate the antioxidant potential of AA to improve common retinal pigment epithelium (ARPE-19) cell viability in the setting of H2O2 induced oxidative stress and evaluate osteoblasts as a potential source of antioxidant recycling.

Methods : In vitro evaluation was performed by incubating ARPE-19 in culture media containing .2mM H2O2 with and without 100uM AA. MTT assay was performed to assess for cell viability. Osteoblast antioxidant recycling potential was tested by exposing MG-63 osteosarcoma cells to culture media containing 100uM DHA. At each time point from 0 to 80 minutes, media was collected and concentrations of DHA and AA were assessed using HPLC. Statistical comparisons were performed using a student’s t-test.

Results : AA successfully attenuated the toxic effects of H2O2, with 88% of ARPE-19 cells remaining viable after exposure to both H2O2and AA, compared to 61% viable after incubation with H2O2 alone (P < .001). Osteoblast antioxidant recycling of DHA was observed with an increase of AA concentration and a concomitant decrease in DHA levels over time. At 80 minutes, the concentration of AA had a 2-fold increase with a paired 2-fold decrease in DHA levels.

Conclusions : These experiments demonstrate the antioxidant potential of AA to attenuate the effects of oxidative stress and its physiologic importance in managing cellular exposure to reactive oxygen species. Osteoblasts exhibited the potential for antioxidant regeneration of AA outside their biological niche. While preliminary, these results demonstrate the promise of an implantable device that continuously recycles antioxidant, eliminating the need for constant injections.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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