Human corneal epithelial cell response to substrate stiffness

Molladavoodi, Sara; Kwon, Hyock-Ju; Medley, John B.; Gorbet, Maud

doi:10.1016/j.actbio.2014.10.005

Cited by 26 publications

(25 citation statements)

References 67 publications

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“…This has been shown previously for corneal epithelial cells [20,45,46]. Using live cell imaging, we have previously demonstrated that HCECs without stretched and visible actin filaments were not capable of migrating effectively [20]. As opposed to the increased healing observed following exposure to low shear stress, exposing cells to high shear stress prior to wounding did not lead to significant changes in scratch width reduction, which could be explained by the non-uniform cytoskeletal rearrangement observed following high shear stress exposure.…”

Section: Resultssupporting

confidence: 82%

“…The organization of the cytoskeleton components such as actin filaments is a key player in proliferation and migration [29,44]. This has been shown previously for corneal epithelial cells [20,45,46]. Using live cell imaging, we have previously demonstrated that HCECs without stretched and visible actin filaments were not capable of migrating effectively [20].…”

Section: Resultsmentioning

confidence: 70%

“…The cornea is exposed to various mechanical stimuli; matrix stiffness [17] and topography [18,19] have been shown to affect keratocytes and corneal endothelial cells. Furthermore, we previously observed that increased substrate stiffness led to changes in cytoskeletal structure and increased migration speed in corneal epithelial cells [20]. Leonard et al .…”

Section: Introductionmentioning

confidence: 99%

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Corneal epithelial cells exposed to shear stress show altered cytoskeleton and migratory behaviour

et al. 2017

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Cells that form the corneal epithelium, the outermost layer of the cornea, are exposed to shear stress through blinking during waking hours. In this in vitro study, the effect of fluid shear stress on human corneal epithelial cells (HCECs) was investigated. Following exposure to shear stresses of 4 and 8 dyn/cm2, HCECs showed cytoskeletal rearrangement with more prominent, organized and elongated filamentous actin. Cytoskeletal changes were time-dependent, and were most significant after 24 hours of shear stress. Higher rates of migration and proliferation, as evaluated by a scratch assay, were also observed following 24 hours of low shear stress exposure (4 dyn/cm2). This result contrasted the poor migration observed in samples scratched before shear exposure, indicating that shear-induced cytoskeletal changes played a key role in improved wound healing and must therefore precede any damage to the cell layer. HCEC cytoskeletal changes were accompanied by an upregulation in integrin β1 and downregulation of ICAM-1. These results demonstrate that HCECs respond favourably to flow-induced shear stress, impacting their proliferation and migration properties as well as phenotype.

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Section: Resultssupporting

confidence: 82%

Section: Resultsmentioning

confidence: 70%

Section: Introductionmentioning

confidence: 99%

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Corneal epithelial cells exposed to shear stress show altered cytoskeleton and migratory behaviour

et al. 2017

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“…The same group showed that central cornea is stiffer than peripheral cornea, resulting in migration and differentiation of LSCs [29] . Molladavoodi et al supported this finding when demonstrating that human corneal epithelial cells had lower rates of migration in compliant tissues [30] . It has therefore been hypothesised that the mechanical properties (stiffness) of the corneal anterior surface represents a major factor regulating corneal epithelium homeostasis [29] .…”

Section: Introductionmentioning

confidence: 90%

Gamma-irradiated human amniotic membrane decellularised with sodium dodecyl sulfate is a more efficient substrate for the ex vivo expansion of limbal stem cells

et al. 2017

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“…Epithelial and endothelial cells are mechanically regulated, with ECM stiffness known to modulate cytoskeletal dynamics 15,16 , proliferation 17 , transcriptional regulation 18 , epithelial-mesenchymal transition (EMT) [19][20][21] and metastasis 22,23 . The principal function of these cell types is to separate luminal spaces from underlying tissues by forming cohesive barriers, with barrier function largely imparted by cell-cell adhesion mediated by cellular junctions [24][25][26] .…”

Section: Introductionmentioning

confidence: 99%

Development of anin vitroExperimental Model for Investigating the Effect of Matrix Stiffness on Epithelial Barrier Permeability

Roy

Turner-Brannen

West

2019

Preprint

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Epithelial cells are well-known to be modulated by extracellular mechanical factors including substrate stiffness. However, the effect of substrate stiffness on an epithelial cell's principal function -creating an effective barrier to protect the underlying tissue -cannot be directly measured using existing experimental techniques. We developed a strategy involving ethylenediamine aminolysis and glutaraldehyde crosslinking to chemically graft polyacrylamide hydrogels with tunable stiffness to PET Transwell membranes. Grafting success was evaluated using spectroscopic methods, scrape tests, and extended incubation in culture. By assessing apical to basolateral transfer of fluorescent tracers, we demonstrated that our model is permeable to biologically relevant molecules and usable for direct measurement of barrier function by calculating paracellular permeability.We found that BEAS-2B epithelial cells form a more effective barrier on stiff substrates, likely attributable to increased cell spreading. We also observed barrier impairment after treatment with transforming growth factor beta, indicating loss of cell-cell junctions, validating our model's ability to detect biologically relevant stimuli. Thus, we have created an experimental model that allows explicit measurement of epithelial barrier function for cells grown on different substrate stiffnesses. This model will be a valuable tool to study mechanical regulation of epithelial and endothelial barrier function in health and disease.

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Human corneal epithelial cell response to substrate stiffness

Cited by 26 publications

References 67 publications

Corneal epithelial cells exposed to shear stress show altered cytoskeleton and migratory behaviour

Corneal epithelial cells exposed to shear stress show altered cytoskeleton and migratory behaviour

Gamma-irradiated human amniotic membrane decellularised with sodium dodecyl sulfate is a more efficient substrate for the ex vivo expansion of limbal stem cells

Development of anin vitroExperimental Model for Investigating the Effect of Matrix Stiffness on Epithelial Barrier Permeability

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