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Effects of PEG-based thermoresponsive polymer brushes on fibroblast spreading and gene expression

: Sefcik, L.S.; Kaminski, A.; Ling, K.; Laschewsky, A.; Lutz, J.-F.; Wischerhoff, E.


Cellular and molecular bioengineering 6 (2013), Nr.3, S.287-298
ISSN: 1865-5025
ISSN: 1865-5033
Fraunhofer IAP ()

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 cycli ng 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.