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Influence of Inherent Mechanophenotype on Competitive Cellular Adherence

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Abstract

Understanding the role of mechanophenotype in competitive adherence of cells to other cells versus underlying substrates can inform such processes as tissue development, cancer progression, and wound healing. This study investigated how mechanophenotype, defined by whole-cell, elastic/viscoelastic properties for the perinuclear region, and cellular assembly are intertwined through the mechanosensing process. Atomic force microscopy was used to characterize the temporal elastic/viscoelastic properties of individual and assembled fibroblasts grown on substrates with elastic moduli above, below, or similar to whole-cell mechanophenotypes measured for three, genetically modified cell lines. All cells were at their most compliant immediately after plating but transitioned to distinct, stiffer mechanophenotypes by Day 1 after acclimation. This mechanical state, and cellular assembly/morphology, did not change significantly over the following three days of testing, regardless of substrate compliance or cellular organization (multi-cell nodules/plaques or single cells). Interestingly, cells formed 3D nodules when attached to substrates with elastic moduli less than their own but spread readily on substrates with moduli equal to or greater than their own, suggesting a preference to adhere to the stiffest surface sensed (substrate or cell). This suggests that inherent mechanophenotype plays a role as a competing surface during microenvironment mechanosensing and subsequent cell–cell-substrate organization.

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Abbreviations

AFM:

Atomic force microscopy

COL-I:

Collagen type-1

dnRhoA:

dnRhoA (T19 N) - GFP transfected WI-38 cells

ECM:

Extracellular matrix

E elastic :

Elastic modulus

E 0 :

Instantaneous modulus

E R :

Relaxed modulus

FN:

Fibronectin

LN:

Laminin

PAAm:

Polyacrylamide

WI-38:

WI-38 VA-13 subline 2RA

β-Actin:

β-Actin-GFP transfected WI-38 cells

µ :

Apparent viscosity

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Acknowledgments

The authors would like to thank Louise E. O. Darling for the GFP plasmids and Jessica S. Sadick and Vera Fonseca for help with western blots. This work was supported by awards from the National Institute of Health (R01 AR063642, P20 GM104937) and the National Science Foundation (CAREER CBET 1253189). The content of this article is solely the responsibility of the authors and does not necessarily represent the official views of the National Science Foundation or National Institutes of Health.

Conflict of Interest

Manisha K. Shah, Iris H. Garcia-Pak, and Eric M. Darling declare that they have no conflicts of interest.

Data Accessibility

AFM and nodule quantification data can be downloaded from 10.6084/m9.figshare.4341887.

Author information

Authors and Affiliations

  1. Center for Biomedical Engineering, Brown University, 175 Meeting Street, Box G-B397, Providence, RI, USA

    Manisha K. Shah, Iris H. Garcia-Pak & Eric M. Darling

  2. Department of Molecular Pharmacology, Physiology, and Biotechnology, Brown University, Providence, RI, USA

    Eric M. Darling

  3. Department of Orthopaedics, Brown University, Providence, RI, USA

    Eric M. Darling

  4. School of Engineering, Brown University, Providence, RI, USA

    Eric M. Darling

Authors
  1. Manisha K. Shah
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Contributions

Author contributions

M.K.S. and E.M.D. designed the study, analyzed data, and wrote the manuscript. M.K.S. performed all AFM experiments and all experiments with non-transfected and transfected WI-38 cells lines. I.H.G.-P performed MG-63 and SH-SY5Y cellular assembly experiments. All authors gave final approval for publication.

Corresponding author

Correspondence to Eric M. Darling.

Additional information

Associate Editor Michael S. Detamore oversaw the review of this article.

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Shah, M.K., Garcia-Pak, I.H. & Darling, E.M. Influence of Inherent Mechanophenotype on Competitive Cellular Adherence. Ann Biomed Eng 45, 2036–2047 (2017). https://doi.org/10.1007/s10439-017-1841-5

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  • DOI: https://doi.org/10.1007/s10439-017-1841-5

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