Biomechanical strain as a trigger for pore formation in Schlemm's canal endothelial cells

Sietse T. Braakman, Ryan M. Pedrigi, A. Thomas Read, James A.E. Smith, W. Daniel Stamer, C. Ross Ethier, Darryl R. Overby

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

The bulk of aqueous humor passing through the conventional outflow pathway must cross the inner wall endothelium of Schlemm's canal (SC), likely through micron-sized transendothelial pores. SC pore density is reduced in glaucoma, possibly contributing to obstructed aqueous humor outflow and elevated intraocular pressure (IOP). Little is known about the mechanisms of pore formation; however, pores are often observed near dome-like cellular outpouchings known as giant vacuoles (GVs) where significant biomechanical strain acts on SC cells. We hypothesize that biomechanical strain triggers pore formation in SC cells. To test this hypothesis, primary human SC cells were isolated from three non-glaucomatous donors (aged 34, 44 and 68), and seeded on collagen-coated elastic membranes held within a membrane stretching device. Membranes were then exposed to 0%, 10% or 20% equibiaxial strain, and the cells were aldehyde-fixed 5min after the onset of strain. Each membrane contained 3-4 separate monolayers of SC cells as replicates (N=34 total monolayers), and pores were assessed by scanning electron microscopy in 12 randomly selected regions (~65,000μm2 per monolayer). Pores were identified and counted by four independent masked observers. Pore density increased with strain in all three cell lines (p<0.010), increasing from 87±36 pores/mm2 at 0% strain to 342±71 at 10% strain; two of the three cell lines showed no additional increase in pore density beyond 10% strain. Transcellular "I-pores" and paracellular "B-pores" both increased with strain (p<0.038), however B-pores represented the majority (76%) of pores. Pore diameter, in contrast, appeared unaffected by strain (p=0.25), having a mean diameter of 0.40μm for I-pores (N=79 pores) and 0.67μm for B-pores (N=350 pores). Pore formation appears to be a mechanosensitive process that is triggered by biomechanical strain, suggesting that SC cells have the ability to modulate local pore density and filtration characteristics of the inner wall endothelium based on local biomechanical cues. The molecular mechanisms of pore formation and how they become altered in glaucoma may be studied invitro using stretched SC cells.

Original languageEnglish (US)
Pages (from-to)224-235
Number of pages12
JournalExperimental Eye Research
Volume127
DOIs
StatePublished - Oct 2014

Keywords

  • Biomechanics
  • Endothelium
  • Pore
  • Schlemm's canal endothelium
  • Strain
  • Stretch

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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    Braakman, S. T., Pedrigi, R. M., Read, A. T., Smith, J. A. E., Stamer, W. D., Ethier, C. R., & Overby, D. R. (2014). Biomechanical strain as a trigger for pore formation in Schlemm's canal endothelial cells. Experimental Eye Research, 127, 224-235. https://doi.org/10.1016/j.exer.2014.08.003