June 2020
Volume 61, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2020
1α,25-dihydroxy vitamin D3 has a cytoprotective effect against inflammatory stress-induced corneal endothelial cell death
Author Affiliations & Notes
  • Hamid Alemi
    Schepens Eye Research Institute, Somerville, Massachusetts, United States
  • Tomas Blanco
    Schepens Eye Research Institute, Somerville, Massachusetts, United States
  • Rohan Bir Singh
    Schepens Eye Research Institute, Somerville, Massachusetts, United States
  • Hayate Nakagawa
    Schepens Eye Research Institute, Somerville, Massachusetts, United States
  • Zala Luznik
    Schepens Eye Research Institute, Somerville, Massachusetts, United States
  • Jia Yin
    Schepens Eye Research Institute, Somerville, Massachusetts, United States
  • Sunil Chauhan
    Schepens Eye Research Institute, Somerville, Massachusetts, United States
  • Reza Dana
    Schepens Eye Research Institute, Somerville, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Hamid Alemi, None; Tomas Blanco, None; Rohan Singh, None; Hayate Nakagawa, None; Zala Luznik, None; Jia Yin, None; Sunil Chauhan, None; Reza Dana, None
  • Footnotes
    Support  T32-EY007145 (HA) R01-EY012963 (RD) P30-EY003790 (Core)
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 1462. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Hamid Alemi, Tomas Blanco, Rohan Bir Singh, Hayate Nakagawa, Zala Luznik, Jia Yin, Sunil Chauhan, Reza Dana; 1α,25-dihydroxy vitamin D3 has a cytoprotective effect against inflammatory stress-induced corneal endothelial cell death. Invest. Ophthalmol. Vis. Sci. 2020;61(7):1462.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Loss of corneal endothelial cell (CEnC) viability is the predominant cause of graft failure following corneal transplantations. The role of 1α,25-dihydroxy vitamin D3 (VitD) present in the aqueous humor on CEnC integrity is yet to be elucidated. In this study, we investigated the cytoprotective function of VitD on CEnC against inflammatory stress-induced cell death using an ex-vivo murine corneal model.

Methods : Murine cornea cups (n=5/group) were harvested from C57BL/6 mice and incubated in RPMI+10% BSA media in presence of IFN-γ (100 ng/mL) and TNF-α (100 ng/mL) to induce inflammatory stress in CEnC, with or without VitD (10-11 M, 10-9 M or 10-7 M) for 12 hours. The tight junction protein-1 was stained with FITC-conjugated anti ZO-1 antibody. The CEnC apoptosis was evaluated using the TUNEL assay. The images were acquired by using laser scanner confocal microscopy and subsequently analyzed using ImageJ. The frequency of CEnC apoptosis (TUNEL+ vs. ZO-1+) and density of ZO-1+ cells were compared in groups incubated with different concentrations of VitD.

Results : The corneal cups incubated with IFN-γ and TNF-α had a significantly higher frequency of CEnC apoptosis compared to corneal cups that were not exposed to inflammatory stress (6.85±1.39% vs. 1.81±1.39%, p<0.001). CEnC apoptosis was significantly reduced following induction of inflammatory stress upon supplementation the media with 10-11 M VitD (3.75±0.96%, p=0.003), 10-9 M (2.69±0.95%, p<0.001) and 10-7 M (1.86±0.83%, p<0.001) of VitD, in comparison to the non-treated controls (6.85±1.39%).

Conclusions : Our results demonstrate that CEnC death induced by inflammatory stress is significantly reduced by VitD in a dose-dependent fashion. Notably, VitD treatment even at the lowest dose, which approximates its natural concentration in the aqueous humor, resulted in a significant reduction in CEnC death. These findings indicate a possible cytoprotective effect of VitD on corneal endothelial cells.

This is a 2020 ARVO Annual Meeting abstract.

×
×

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.

×