Cell sheet detachment affects the extracellular matrix: A surface science study comparing thermal liftoff, enzymatic, and mechanical methods

Citation

Canavan, H. E.; Cheng, X. H.; Graham, D. J.; Ratner, B. D.; & Castner, D. G. (2005). Cell sheet detachment affects the extracellular matrix: A surface science study comparing thermal liftoff, enzymatic, and mechanical methods. Journal of Biomedical Materials Research Part A, 75A(1), 1-13.

Abstract

This work compares the removal of bovine aortic endothelial cell (BAEC) monolayers via 1) low-temperature liftoff from a "smart polymer," plasma polymerized poly(Nisopropyl acrylamide) (ppNIPAM), 2) enzymatic digestion, and 3) mechanical dissociation from ppNIPAM surfaces. We examine the surfaces after cell removal by using X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectrometry (ToF-SIMS), immunostaining, and cell adhesion assay. Immunoassay results indicate that low-temperature liftoff nonclestructively harvests the cell sheet and most of the underlying extracellular matrix (ECM), whereas enzymatic digestion and mechanical dissociation are damaging to both the cells and ECM. XPS results indicate that amide and alcohol groups attributed to proteins in the ECM are present on postliftoff surfaces. Principal component analysis (PCA) of ToF-SIMS data indicates that molecular ion fragments of amino acids are present on postliftoff surfaces. Finally, a cell adhesion assay seeding new cells on surfaces from which an initial layer of cells was removed via each of the three methods indicates that liftoff and mechanical dissociation leave behind surfaces that better promote cell adhesion. We conclude that the removal of BAEC cells via low-temperature liftoff from ppNIPAM-treated surfaces is less damaging to the ECM proteins remaining at the surface than the other methods. (c) 2005 Wiley Periodicals, Inc.

Keyword(s)

cell removal
ecm
endothelial-cells
in-vitro
ion mass-spectrometry
n-isopropyl acrylamide
n-isopropylacrylamide gels
principal component analysis
responsive culture surfaces
self-assembled monolayers
smooth-muscle-cells
thermoresponsive polymer
tof-sims
volume phase-transition
xps

Notes

963PP
Times Cited:78
Cited References Count:61

Reference Type

Journal Article

Secondary Title

Journal of Biomedical Materials Research Part A

Author(s)

Canavan, H. E.
Cheng, X. H.
Graham, D. J.
Ratner, B. D.
Castner, D. G.

Year Published

2005

Date Published

1759276800

Volume Number

75A

Issue Number

1

Pages

1-13

ISSN/ISBN

1549-3296

DOI

Doi 10.1002/Jbm.A.30297

Times Cited:78
Cited References Count:61 SN - 1549-3296 T1 - Cell sheet detachment affects the extracellular matrix: A surface science study comparing thermal liftoff, enzymatic, and mechanical methods VL - 75A PY - 2005 IS - 1 SP - 1 EP - 13 PY - 2025/10// ER - ">