Cellular and Molecular Bioengineering

, Volume 6, Issue 3, pp 253–265 | Cite as

Stiffness Increases Mononuclear Cell Transendothelial Migration

Article

Abstract

Cardiovascular diseases, including hypertension and the evolution of an atherosclerotic plaque alter the mechanics of the sub-endothelium. The extent to which either arterial softening or stiffening exacerbates or reduces mononuclear leukocyte infiltration from the bloodstream into the wall remains unclear. Mononuclear cell (MNC) transmigration was observed using an in vitro model of the inflamed human vascular endothelium on variable substrate stiffness. Briefly, human aortic endothelial cells were allowed to grow to confluency on gels of 1, 3, 5, 280 kPa, and on glass (~70 GPa), and then activated with TNF-α. Isolated human MNCs were allowed to transmigrate across the inflamed endothelium for 1 h. Fewer MNC transmigrated on soft compared to stiff substrates, yet the relative expression of ICAM-1 and VCAM-1, and the fraction of MNCs that become activated (or changed shape) did not change. Following MNC transendothelial migration the distribution of VCAM-1 is translocated from the apical to basal plasma membrane as revealed through immunofluorescence; since less MNCs transmigrate on soft subendothelial substrates, translocation of VCAM-1 was not observed. These results herein highlight that stiffer subendothelial substrates actually increase MNC transmigration, yet transmigration on stiff substrates can be abrogated by blocking ICAM-1.

Keywords

Human Subendothelial stiffness Cell activation ICAM-1 VCAM-1 Monocytes HAEC In vitro Diapedese Actin 

Abbreviations

MNC

Mononuclear cell

HAEC

Human aortic endothelial cell

TEM

Transendothelial migration

MFI

Mean fluorescent intensity

a.u.

Arbitrary units

PMN

Polymorphonuclear cell

Supplementary material

12195_2013_284_MOESM1_ESM.docx (35 kb)
Supplementary material 1 (DOCX 36 kb)
12195_2013_284_MOESM2_ESM.mov (2.2 mb)
Supplementary material 2 (MOV 2276 kb)
12195_2013_284_MOESM3_ESM.mov (2.2 mb)
Supplementary material 3 (MOV 2268 kb)

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Copyright information

© Biomedical Engineering Society 2013

Authors and Affiliations

  1. 1.Fischell Department of BioengineeringUniversity of MarylandCollege ParkUSA

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