An advanced in vitro model to assess glaucoma onset

S.C. Sacc, S. Tirendi, S. Scarji, M. Fassalacqua, F. Oddone, C.K. Traverso, S. Vernazza, A.M. Bassi

Research output: Contribution to journalArticlepeer-review

Abstract

Glaucoma is the second leading cause of blindness worldwide. Currently, glaucoma treatments aim to lower intraocular pressure by decreasing aqueous humor production or increasing aqueous humor outflow through pharmacological approaches or trabeculectomy. The lack of an effective cure requires new therapeutic strategies. We compared the biological responses of a three-dimensional trabecular meshwork model with or without perfusion bioreactor technology to better understand the early molecular changes induced by prolonged oxidative stress conditions induced by repeated daily peroxide exposure. We used standard 3D cultures of trabecular meshwork cells in Matrigel cultured under either static and dynamic conditions for one week. We studied changes in F-actin expression and organization in the cells, cellular metabolic activity, proinflammatory gene expression, expression of pro- and anti-apoptotic proteins, PARP-1 cleavage, and NFKB activation in the model. We demonstrate that the dynamic conditions improve the adaptive behavior of 3D trabecular meshwork cultures to chronic oxidative stress via offsetting pathway activation. © 2020 ALTEX Edition. All rights reserved.
Original languageEnglish
Pages (from-to)265-274
Number of pages10
JournalAltex
Volume37
Issue number2
DOIs
Publication statusPublished - 2020

Keywords

  • caspase 3
  • collagen type 1
  • collagen type 1 alpha 1
  • complementary DNA
  • cytochrome c
  • gelatinase B
  • immunoglobulin enhancer binding protein
  • interleukin 1alpha
  • interleukin 1beta
  • interleukin 6
  • interstitial collagenase
  • matrigel
  • nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase 1
  • peroxide
  • protein Bax
  • protein bcl 2
  • protein p53
  • second mitochondrial activator of caspase
  • stromelysin
  • synaptotagmin I
  • transforming growth factor beta2
  • tumor necrosis factor
  • ubiquitin C
  • unclassified drug
  • ubiquitin
  • antiapoptotic activity
  • apoptosis assay
  • Article
  • chronic stress
  • gene expression
  • glaucoma
  • human
  • human cell
  • immunoreactivity
  • in vitro study
  • metabolic activity assay
  • oxidative stress
  • polyacrylamide gel electrophoresis
  • protein expression
  • real time polymerase chain reaction
  • resazurin assay
  • RNA extraction
  • three dimensional cell culture
  • trabecular meshwork
  • Western blotting
  • adaptive behavior
  • cell culture technique
  • cell growth
  • confocal microscopy
  • controlled study
  • cytokine production
  • down regulation
  • flow cytometry
  • genetic transfection
  • signal transduction
  • upregulation

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