June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Melatonin protects mitochondria in Non-AMD cybrids and AMD cybrids
Author Affiliations & Notes
  • Farid Jose Thomaz Neto
    Ophthalmology, Gavin Herbert Eye Institute, Irvine, California, United States
    Ophthalmology, University of California Irvine, Irvine, California, United States
  • Nasim Salimiagdam
    Ophthalmology, Gavin Herbert Eye Institute, Irvine, California, United States
    Ophthalmology, University of California Irvine, Irvine, California, United States
  • Baruch D Kuppermann
    Ophthalmology, Gavin Herbert Eye Institute, Irvine, California, United States
    Ophthalmology, University of California Irvine, Irvine, California, United States
  • M.Cristina Kenney
    Ophthalmology, Gavin Herbert Eye Institute, Irvine, California, United States
    Ophthalmology, University of California Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Farid Thomaz Neto None; Nasim Salimiagdam None; Baruch Kuppermann Allegro Ophthalmics, LLC; Allergan; Aprea Therapeutics, Inc; Cell Care Therapeutics; DOSE Medical Corporation; Eyedaptic; Galimedix; Genentech, Inc; Glaukos Corporation; Interface Biologics; IVERIC Bio/Ophthotech Corporation; jCyte; Novartis Pharmaceuticals Corporation; Ocunexus Therapeutics; Regeneron Pharmaceuticals, Inc; ReVana Therapeutics; Ripple Therapeutics; Theravance Biopharma, Code C (Consultant/Contractor); M.Cristina Kenney None
  • Footnotes
    Support  Discovery Eye Foundation, Polly and Michael Smith, Iris and the B. Gerald Cantor Foundation, Beckman Initiative for Macular Research, National Institute on Aging [AG006168 to SMJ] and National Eye Institute R01 EY027363 (MCK). We acknowledge the support of the Institute for Clinical and Translational Science (ICTS) at University of California, Irvine. Supported in part by an Unrestricted Departmental Grant from Research to Prevent Blindness.
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 787 – F0346. doi:
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    • Get Citation

      Farid Jose Thomaz Neto, Nasim Salimiagdam, Baruch D Kuppermann, M.Cristina Kenney; Melatonin protects mitochondria in Non-AMD cybrids and AMD cybrids. Invest. Ophthalmol. Vis. Sci. 2022;63(7):787 – F0346.

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

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Abstract

Purpose : Among elderly people in developed countries age-related macular degeneration (AMD) is the leading cause of progressive vision loss. Hallmark features of AMD are an accumulation of drusen in the Bruch’s membrane and dysfunction of retinal pigment epithelial (RPE) cells. Oxidative stress is an important factor contributing to the damaged RPE cells. Multiple studies provide evidence that melanin is an important antioxidant factor which leads to reduction of the reactive oxygen species (ROS). Melatonin also has anti-inflammatory, antitumor, and antiangiogenic properties.
Evaluate melatonin protection in Non-AMD cybrids and in AMD cybrids.

Methods : AMD patients and age-matched non-AMD individuals provided platelets that were combined with mitochondria-deficient (Rho0) ARPE-19 cells. The resultant Cybrid cell lines have identical nuclei but mitochondria from different individuals. Cybrids were cultured in 96-well plates (10,000 cells/well) and treated with Melatonin 200 µM for 24 hours. Ethanol was used as vehicle-control. ROS levels and cellular metabolism were measured by the H2DCFDA and MTT assays, respectively. After adding 100 µl H2DCFDA solution/well, plates were read via the fluorescent plate reader with the excitation (EX, 492nm) and emission (EM, 520nm) wavelengths. For cell metabolism, 10 µl of MTT assay reagent was added per well and plates were incubated in 37°C for 3 hours. After adding 100 μl/well of DMSO, plates were analyzed (signal at 570 nm and reference at 630 nm) with the absorbance plate reader. – Statistics were calculated using Student T-test.

Results : ROS levels of non-AMD and AMD cybrids with melatonin declined to 90.50 ± 3.79 (P< 0.0001, n=4) and 84.50 ± 1.55 (P< 0.0001, n=4) versus Ethanol groups (100.8 ± 0.478, n=4) and (99.75 ± 1.75, n=4), respectively.
MTT Assay levels of non-AMD and AMD cybrids with Melatonin improved to 129 ± 2.97 (P< 0.0001, n=4) and 132.3 ± 2.78 (P< 0.0001, n=4) versus Ethanol groups (100.8 ±0.478, n=4) and (99.75 ±1.75, n=4), respectively.

Conclusions : Non-AMD cybrids and AMD cybrids treated with melatonin had significantly increased cellular metabolism and lower levels of ROS, demonstrating that melatonin could protect cells against oxidative stress.

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

 

 

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