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

This work compares the removal of bovine aortic endothelial cell (BAEC) monolayers via 1) low‐temperature liftoff from a “smart polymer,” plasma polymerized poly(N‐isopropyl acrylamide) (ppNIPAM), 2) enzymatic digestion, and 3) mechanical dissociation from ppNIPAM surfaces. We examine the surfaces a...

Full description

Saved in:
Bibliographic Details
Published in:Journal of biomedical materials research. Part A 2005-10, Vol.75A (1), p.1-13
Main Authors: Canavan, Heather E., Cheng, Xuanhong, Graham, Daniel J., Ratner, Buddy D., Castner, David G.
Format: Article
Language:English
Subjects:
Acrylamides - chemistry
Animals
Aorta - cytology
Biocompatible Materials - chemistry
Biophysics - methods
Cattle
Cell Adhesion
cell removal
Cells, Cultured
ECM
Endothelial Cells - cytology
Enzymes - chemistry
Extracellular Matrix - metabolism
Immunoassay
N-isopropyl acrylamide
Polymers - chemistry
Principal Component Analysis
Spectrometry, X-Ray Emission
Stress, Mechanical
Surface Properties
Temperature
Time Factors
ToF-SIMS
XPS
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:This work compares the removal of bovine aortic endothelial cell (BAEC) monolayers via 1) low‐temperature liftoff from a “smart polymer,” plasma polymerized poly(N‐isopropyl 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 nondestructively 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. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005
ISSN:1549-3296
1552-4965
DOI:10.1002/jbm.a.30297