Cytoskeletal networks in an epithelial sheet: Mechanical characterization using composite diaphragm inflation

John C. Selby, Stephen A. Boppart, Mark A. Shannon

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

A new experimental methodology combining concepts of classical mechanical testing with basic microfabrication techniques has been developed to investigate the mechanical architecture of cytoskeletal networks and anchoring junctions within a monolayer of normal human epidermal keratinocytes (NHEKs) before, during, and after the in vitro formation of an epithelial sheet. Referred to as composite diaphragm inflation (CDI), the method resembles a microscale model of a classical axisvmmetric biaxial membrane inflation experiment. CDI utilizes a free-standing polydimethylsiloxane (PDMS) elastomer membrane as a substrate for the culture of NHEKs into an epithelial sheet via calcium induction of cytoskeletal network reorganization and anchoring junction assembly. Together, the NHEK monolayer and the PDMS membrane form a composite diaphragm suitable for mechanical measurements. If the geometry and elastic properties of the PDMS membrane are scaled appropriately, then the prestress and apparent stiffness of the NHEK layer can be characterized by measured deviations in the load-displacement behavior of the composite diaphragm with respect to the behavior of the PDMS membrane measured prior to cell culture.

Original languageEnglish (US)
Title of host publication2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology
Pages198-200
Number of pages3
DOIs
StatePublished - Dec 1 2005
Event2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology - Oahu, HI, United States
Duration: May 12 2005May 15 2005

Publication series

Name2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology
Volume2005

Other

Other2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology
CountryUnited States
CityOahu, HI
Period5/12/055/15/05

Fingerprint

Diaphragms
Polydimethylsiloxane
Membranes
Composite materials
Monolayers
Mechanical testing
Microfabrication
Cell culture
Elastomers
Calcium
Stiffness
Geometry
Keratinocytes
Substrates
Experiments

Keywords

  • Anchoring junctions
  • Cell mechanics
  • Cytoskeleton

ASJC Scopus subject areas

  • Engineering(all)

Cite this

Selby, J. C., Boppart, S. A., & Shannon, M. A. (2005). Cytoskeletal networks in an epithelial sheet: Mechanical characterization using composite diaphragm inflation. In 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology (pp. 198-200). [1548425] (2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology; Vol. 2005). https://doi.org/10.1109/MMB.2005.1548425

Cytoskeletal networks in an epithelial sheet : Mechanical characterization using composite diaphragm inflation. / Selby, John C.; Boppart, Stephen A.; Shannon, Mark A.

2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology. 2005. p. 198-200 1548425 (2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology; Vol. 2005).

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Selby, JC, Boppart, SA & Shannon, MA 2005, Cytoskeletal networks in an epithelial sheet: Mechanical characterization using composite diaphragm inflation. in 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology., 1548425, 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology, vol. 2005, pp. 198-200, 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology, Oahu, HI, United States, 5/12/05. https://doi.org/10.1109/MMB.2005.1548425
Selby JC, Boppart SA, Shannon MA. Cytoskeletal networks in an epithelial sheet: Mechanical characterization using composite diaphragm inflation. In 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology. 2005. p. 198-200. 1548425. (2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology). https://doi.org/10.1109/MMB.2005.1548425
Selby, John C. ; Boppart, Stephen A. ; Shannon, Mark A. / Cytoskeletal networks in an epithelial sheet : Mechanical characterization using composite diaphragm inflation. 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology. 2005. pp. 198-200 (2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology).
@inproceedings{780621ebc9cd464c881e6bdd22a5d331,
title = "Cytoskeletal networks in an epithelial sheet: Mechanical characterization using composite diaphragm inflation",
abstract = "A new experimental methodology combining concepts of classical mechanical testing with basic microfabrication techniques has been developed to investigate the mechanical architecture of cytoskeletal networks and anchoring junctions within a monolayer of normal human epidermal keratinocytes (NHEKs) before, during, and after the in vitro formation of an epithelial sheet. Referred to as composite diaphragm inflation (CDI), the method resembles a microscale model of a classical axisvmmetric biaxial membrane inflation experiment. CDI utilizes a free-standing polydimethylsiloxane (PDMS) elastomer membrane as a substrate for the culture of NHEKs into an epithelial sheet via calcium induction of cytoskeletal network reorganization and anchoring junction assembly. Together, the NHEK monolayer and the PDMS membrane form a composite diaphragm suitable for mechanical measurements. If the geometry and elastic properties of the PDMS membrane are scaled appropriately, then the prestress and apparent stiffness of the NHEK layer can be characterized by measured deviations in the load-displacement behavior of the composite diaphragm with respect to the behavior of the PDMS membrane measured prior to cell culture.",
keywords = "Anchoring junctions, Cell mechanics, Cytoskeleton",
author = "Selby, {John C.} and Boppart, {Stephen A.} and Shannon, {Mark A.}",
year = "2005",
month = "12",
day = "1",
doi = "10.1109/MMB.2005.1548425",
language = "English (US)",
isbn = "0780387112",
series = "2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology",
pages = "198--200",
booktitle = "2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology",

}

TY - GEN

T1 - Cytoskeletal networks in an epithelial sheet

T2 - Mechanical characterization using composite diaphragm inflation

AU - Selby, John C.

AU - Boppart, Stephen A.

AU - Shannon, Mark A.

PY - 2005/12/1

Y1 - 2005/12/1

N2 - A new experimental methodology combining concepts of classical mechanical testing with basic microfabrication techniques has been developed to investigate the mechanical architecture of cytoskeletal networks and anchoring junctions within a monolayer of normal human epidermal keratinocytes (NHEKs) before, during, and after the in vitro formation of an epithelial sheet. Referred to as composite diaphragm inflation (CDI), the method resembles a microscale model of a classical axisvmmetric biaxial membrane inflation experiment. CDI utilizes a free-standing polydimethylsiloxane (PDMS) elastomer membrane as a substrate for the culture of NHEKs into an epithelial sheet via calcium induction of cytoskeletal network reorganization and anchoring junction assembly. Together, the NHEK monolayer and the PDMS membrane form a composite diaphragm suitable for mechanical measurements. If the geometry and elastic properties of the PDMS membrane are scaled appropriately, then the prestress and apparent stiffness of the NHEK layer can be characterized by measured deviations in the load-displacement behavior of the composite diaphragm with respect to the behavior of the PDMS membrane measured prior to cell culture.

AB - A new experimental methodology combining concepts of classical mechanical testing with basic microfabrication techniques has been developed to investigate the mechanical architecture of cytoskeletal networks and anchoring junctions within a monolayer of normal human epidermal keratinocytes (NHEKs) before, during, and after the in vitro formation of an epithelial sheet. Referred to as composite diaphragm inflation (CDI), the method resembles a microscale model of a classical axisvmmetric biaxial membrane inflation experiment. CDI utilizes a free-standing polydimethylsiloxane (PDMS) elastomer membrane as a substrate for the culture of NHEKs into an epithelial sheet via calcium induction of cytoskeletal network reorganization and anchoring junction assembly. Together, the NHEK monolayer and the PDMS membrane form a composite diaphragm suitable for mechanical measurements. If the geometry and elastic properties of the PDMS membrane are scaled appropriately, then the prestress and apparent stiffness of the NHEK layer can be characterized by measured deviations in the load-displacement behavior of the composite diaphragm with respect to the behavior of the PDMS membrane measured prior to cell culture.

KW - Anchoring junctions

KW - Cell mechanics

KW - Cytoskeleton

UR - http://www.scopus.com/inward/record.url?scp=33845336542&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33845336542&partnerID=8YFLogxK

U2 - 10.1109/MMB.2005.1548425

DO - 10.1109/MMB.2005.1548425

M3 - Conference contribution

AN - SCOPUS:33845336542

SN - 0780387112

SN - 9780780387119

T3 - 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology

SP - 198

EP - 200

BT - 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology

ER -