2D- and 3D-cultures of human trabecular meshwork cells: A preliminary assessment of an in vitro model for glaucoma study

PLoS One. 2019 Sep 6;14(9):e0221942. doi: 10.1371/journal.pone.0221942. eCollection 2019.

Abstract

A physiologically relevant in vitro human-based model could be the 'gold standard' to clarify the pathological steps involved in glaucoma onset. In this regard, human 3D cultures may represent an excellent starting point to achieve this goal. Indeed, the 3D matrix allows to re-create the in vivo-like tissue architecture, maintaining its functionality and cellular behaviour, compared to the 2D model. Thus, we propose a comparison between the 2D and 3D in vitro models of human trabecular meshwork cells in terms of cellular responses after chronic stress exposure. Our results showed that 3D-cells are more sensitive to intracellular reactive oxidative specie production induced by hydrogen peroxide treatment, compared to 2D cultures. Additionally, in 3D cultures a more accurate regulation of the apoptosis trigger and cell adaptation mechanisms was detected than in 2D models. In line with these findings, the 3D-HTMC model shows the ability to better mimic the in vivo cell behaviour in adaptive responses to chronic oxidative stress than 2D.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects
  • Cell Culture Techniques
  • Cell Respiration / drug effects
  • Cytokines / metabolism
  • Gene Expression Regulation / drug effects
  • Glaucoma / pathology*
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • NF-kappa B / genetics
  • Oxidative Stress / drug effects
  • Reactive Oxygen Species / metabolism
  • Trabecular Meshwork / cytology*
  • Trabecular Meshwork / drug effects
  • Trabecular Meshwork / metabolism
  • Transcriptional Activation / drug effects

Substances

  • Cytokines
  • NF-kappa B
  • Reactive Oxygen Species
  • Hydrogen Peroxide

Grants and funding

Dr. Francesco Oddone and Dr. Stefania Vernazza were supported by Italian Ministry of Health and Fondazione Roma, Rome, Italy. This work was supported by Omikron Italia srl, Rome, Italy to AMB. SCS and AMB received the award from the Italian Association for Glaucoma Study (A.I.S.G.) – Omikron Italia 2017 “Marco Centofanti Neuroprotection and Glaucoma”. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.