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A Rapid, Scalable Method for the Isolation, Functional Study, and Analysis of Cell-derived Extracellular Matrix
Published on: January 4, 2017
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Characterization of extracellular matrix deposited by segmental trabecular meshwork cells
VijayKrishna Raghunathan1, Andrews Nartey1, Kamesh Dhamodaran1
1Department of Basic Sciences, College of Optometry, University of Houston, Houston, TX, USA.
Experimental Eye Research
|July 28, 2023
Summary
Trabecular meshwork cells from low-flow regions create stiffer extracellular matrices with more crosslinks, especially when exposed to steroids. This suggests distinct cellular identities influence matrix properties in glaucoma research.
Area of Science:
- Biochemistry
- Biophysics
- Cell Biology
Background:
- Extracellular matrix (ECM) properties significantly impact cell behavior in health and disease.
- Ocular hypertension and glaucoma involve stiffening of the trabecular meshwork (TM) and changes in its ECM (matrisome) in a segmental pattern.
- Previous research indicates ECM alterations in the TM are dynamic, responding to age and pressure, but the reasons for segmental differences remain unclear due to limited in vitro models.
Purpose of the Study:
- To investigate the biomechanical and compositional differences in cell-derived matrices (CDMs) from distinct segmental flow regions of the trabecular meshwork.
- To determine the influence of steroid treatment (dexamethasone) on these matrices.
- To explore potential correlations between matrix stiffness and crosslinking in TM pathobiology.
Main Methods:
- Primary TM cells were isolated from high and low flow regions.
- Cells were cultured for 4 weeks with or without dexamethasone to generate CDMs.
- Atomic force microscopy and mass spectrometry were used to analyze CDM biomechanics, matrisome composition, and crosslink incidence.
Main Results:
- Matrices from low-flow regions were stiffer and contained more immature and mature crosslinks compared to high-flow regions.
- Steroid treatment exacerbated these differences and induced differential responses in high- and low-flow cells.
- No direct correlation was found between elastic moduli and the presence of mature or immature crosslinks in the CDMs.
Conclusions:
- Distinct differences exist in the composition and mechanics of matrices deposited by TM cells from different flow regions, suggesting unique cellular identities and potential for mechanical memory.
- While the direct mechanistic basis for matrix stiffness was not determined, CDMs serve as a valuable tool for studying cell-matrix interactions in TM pathobiology.
- These findings contribute to understanding the complex mechanisms underlying glaucoma and trabecular meshwork dysfunction.
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