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Effects of PEG-Based Thermoresponsive Polymer Brushes on Fibroblast Spreading and Gene Expression

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Abstract

The use of thermoresponsive polymer (TRP) substrates, such as PNIPAM, has become a valuable tool for the gentle detachment of cells from their culture surface while retaining cell–cell and cell–matrix contacts. P(MEO2MA-co-OEGMA) substrates (PMO) are an alternative to PNIPAM and offer a tunable LCST, but remain relatively uncharacterized as cell culture substrates. To evaluate the cell–material interaction on these TRPs, L-929 fibroblasts were cultured on PMO substrates with a LCST of 34 °C and differences in cell attachment, morphology, and gene expression were compared to culture on PNIPAM or tissue culture plastic over 48 h. Since thermal switching of TRP substrates is reversible, the cellular response was also examined as a function of repeated substrate use (i.e., “cycling”). There was an observed lag in cell attachment and spreading over time on PMO substrates that was not observed on PNIPAM surfaces or after repeated cycling. Preconditioning PMO surfaces by cycling in serum-containing media without cells resulted in enhanced initial attachment rates. Gene expression for markers FN1, IL-6, Bcl-2, and Dusp2 were up-regulated on native PMO substrates compared to PNIPAM, but were attenuated by cycling. There were no differences in expression for any gene between native and cycled PNIPAM substrates. Results demonstrate that biocompatibility of PMO surfaces is enhanced with successive use in culture.

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Abbreviations

MEO2MA:

2-(2-Methoxyethoxy)ethyl methacrylate

OEGMA:

Oligo(ethylene glycol) methacrylate

PMO:

P(MEO2MA-co-OEGMA)

LCST:

Lower critical solution temperature

TCP:

Tissue culture plastic

PNIPAM:

Poly(N-isopropylacrylamide)

PEG:

Poly(ethylene glycol)

PEI:

Poly(ethyleneimine)

PSS:

Poly(styrene sulfonate)

PDADMAC:

Poly(diallyl dimethyl ammonium chloride)

PCR:

Polymerase chain reaction

TRP:

Thermoresponsive polymer

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Acknowledgments

We thank Anne Sänger for preparation of the biofunctional coatings, Tyler Fruneaux for help with cellular imaging, and Selin Demirler and Allyson Hill for assistance with data analysis. L.S. acknowledges the National Science Foundation for support through grant #0923273.

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, Lafayette College, Easton, PA, 18042, USA

    Lauren S. Sefcik, Ashley Kaminski & Kevin Ling

  2. Fraunhofer Institute for Applied Polymer Research, Geiselbergstr. 69, 14476, Potsdam-Golm, Germany

    André Laschewsky, Jean-François Lutz & Erik Wischerhoff

  3. Institut Charles Sadron, UPR22 CNRS, 23 rue du Loess, BP 84047, 67034, Strasbourg Cedex 2, France

    Jean-François Lutz

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  1. Lauren S. Sefcik
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  2. Ashley Kaminski
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Correspondence to Lauren S. Sefcik.

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Associate Editor Aleksander S. Popel oversaw the review of this article.

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Sefcik, L.S., Kaminski, A., Ling, K. et al. Effects of PEG-Based Thermoresponsive Polymer Brushes on Fibroblast Spreading and Gene Expression. Cel. Mol. Bioeng. 6, 287–298 (2013). https://doi.org/10.1007/s12195-013-0286-7

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