June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Cellular and molecular changes in human corneal fibroblasts following hydrogen sulfide exposure in vitro
Author Affiliations & Notes
  • Praveen Kumar Balne
    Harry S. Truman Memorial Veterans’ Hospital, Columbia, Missouri, United States
    One-Health Vision Research Program, Departments of Veterinary Medicine & Surgery and Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, United States
  • Nishant Rajiv Sinha
    Harry S. Truman Memorial Veterans’ Hospital, Columbia, Missouri, United States
    One-Health Vision Research Program, Departments of Veterinary Medicine & Surgery and Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, United States
  • Alexandria Caroline Hofmann
    One-Health Vision Research Program, Departments of Veterinary Medicine & Surgery and Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, United States
  • Ratnakar Tripathi
    Harry S. Truman Memorial Veterans’ Hospital, Columbia, Missouri, United States
    One-Health Vision Research Program, Departments of Veterinary Medicine & Surgery and Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, United States
  • Suneel Gupta
    Harry S. Truman Memorial Veterans’ Hospital, Columbia, Missouri, United States
    One-Health Vision Research Program, Departments of Veterinary Medicine & Surgery and Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, United States
  • Prashant R. Sinha
    Harry S. Truman Memorial Veterans’ Hospital, Columbia, Missouri, United States
    One-Health Vision Research Program, Departments of Veterinary Medicine & Surgery and Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, United States
  • Rajiv R Mohan
    Harry S. Truman Memorial Veterans’ Hospital, Columbia, Missouri, United States
    One-Health Vision Research Program, Departments of Veterinary Medicine & Surgery and Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, United States
  • Footnotes
    Commercial Relationships   Praveen Balne, None; Nishant Sinha, None; Alexandria Hofmann, None; Ratnakar Tripathi, None; Suneel Gupta, None; Prashant Sinha, None; Rajiv Mohan, None
  • Footnotes
    Support  Phi Zeta pilot research grant (PKB), and the Ruth M. Kraeuchi Missouri Endowed Chair Ophthalmology University of Missouri Fund (RRM). A partial support was from the NIH/NEI 1R21EY030233 and 1R21EY030234 grants, and the Veterans Health Affairs Merit 1I01BX00357 and RCS grants (RRM)
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 944. doi:
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    • Get Citation

      Praveen Kumar Balne, Nishant Rajiv Sinha, Alexandria Caroline Hofmann, Ratnakar Tripathi, Suneel Gupta, Prashant R. Sinha, Rajiv R Mohan; Cellular and molecular changes in human corneal fibroblasts following hydrogen sulfide exposure in vitro. Invest. Ophthalmol. Vis. Sci. 2021;62(8):944.

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

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Abstract

Purpose : Hydrogen sulfide (H2S) is a common environmental pollutant and a chemical warfare agent. H2S exposure to the eye leads to ocular irritation, pain, and vision loss, however, underlying molecular mechanisms driving its toxicity to the cornea is still unclear. We sought to study changes in cellular and molecular parameters in primary human corneal stromal fibroblasts following H2S exposure in vitro.

Methods : A total of 30 healthy human donor corneas obtained from the eye bank were used in this study. Primary human corneal stromal fibroblasts (hCSFs) were generated from healthy donor corneas following our standard protocol and passages two to five were used for in vitro experiments. Sodium hydrosulfide (NaSH) was used as a source of H2S and hCSF cells were exposed to H2S for up to 72h and the changes in mitochondrial function were assessed using Complex IV (cytochrome c oxidase) Human Specific Activity Microplate Assay Kit (Abcam, Cambridge, MA, USA) according to the manufacturer’s guidelines. The mRNA expression of cell death genes - Receptor-interacting serine/threonine-protein kinase 1 (RIP1), Tumor necrosis factor receptor type 1-associated DEATH domain protein (TRADD), Caspase 8, and Mixed lineage kinase domain-like protein (MLKL) was studied using QuantStudio 6 Flex Real-Time PCR System (Applied Biosystems, CA, USA).

Results : H2S exposure showed dose and duration dependent cytotoxicity to hCSFs in cytotoxicity assays and IC50 of H2S was determined to be 5.35 mM. H2S IC50 exposed hCSFs showed significantly decreased levels of cytochrome c oxidase enzyme activity (p<0.01) and quantity (p<0.0001) compared to healthy cells. The mRNA expression of RIP1, TRADD, Caspase 8, and MLKL genes were significantly altered in H2S IC50 exposed hCSFs compared to control (p≤0.05).

Conclusions : H2S exposure to human corneal fibroblasts activates cell death pathways and damages mitochondrial function in vitro.

This is a 2021 ARVO Annual Meeting abstract.

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