Skip to main content
SpringerLink
Log in

Poly(2-substituted-2-oxazoline) surfaces for dermal fibroblasts adhesion and detachment

  • Published:
Journal of Materials Science: Materials in Medicine Aims and scope Submit manuscript

Abstract

The thermoresponsive surfaces of brush structure (linear polymer chains tethered on the surface) based on poly(2-isopropyl-2-oxazoline)s and copolymers of 2-ethyl-2-oxazoline and 2-nonyl-2-oxazoline were obtained using the grafting-to method. The living oxazoline (co)polymers have been synthesized by cationic ring-opening polymerization and subsequently terminated by the reactive amine groups present on the surface. The changes in the surface morphology, philicity and thickness occurring during surface modification were monitored via atomic force microscopy, contact angle and ellipsometry. The thickness of the (co)poly(2-substituted-2-oxazoline) layers ranged from 4 to 11 nm depending on the molar mass of immobilized polymer and reversibly varied with the temperature changes. This confirmed thermoresponsive properties of obtained surfaces. The obtained polymer surfaces were used as a support for dermal fibroblast culture and detachment. The fibroblasts’ adhesion and proliferation on the polymer surfaces were observed when the culture temperature was above the cloud point temperature of the immobilized polymer. Lowering the temperature resulted in the detachment of the dermal fibroblast sheets from the polymer layers, which makes these surfaces suitable for the treatment of wounds and in skin tissue engineering.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

References

  1. Yamada N, Okano T, Sakai H, Karikusa F, Sawasaki Y, Sakurai Y. Thermo-responsive polymeric surfaces; control of attachment and detachment of cultured cells. Macromol Rapid Commun. 1990;11:571–6.

    Article  Google Scholar 

  2. Elloumi-Hannachi I, Yamato M, Okano T. Cell sheet engineering: a unique nanotechnology for scaffold-free tissue reconstruction with clinical applications in regenerative medicine. J Intern Med. 2009;267:54–70.

    Article  Google Scholar 

  3. Kikuchi A, Okano T. Nanostructured designs of biomedical materials: applications of cell sheet engineering to functional regenerative tissues and organs. J Control Release. 2005;101:69–84.

    Article  Google Scholar 

  4. Nagase K, Kobayashi J, Okano T. Temperature-responsive intelligent interfaces for biomolecular separation and cell sheet engineering. J R Soc Interface. 2009;6:S293–309.

    Article  Google Scholar 

  5. Ebara M, Yamato M, Hirose M, Aoyagi T, Kikuchi A, Sakai K, Okano T. Copolymerization of 2-carboxyisopropylacrylamide with N-isopropylacrylamide accelerates cell detachment from grafted surfaces by reducing temperature. Biomacromolecules. 2003;4:344–9.

    Article  Google Scholar 

  6. Ebara M, Yamato M, Nagai S, Aoyagi T, Kikuchi A, Sakai K, Okano T. Incorporation of new carboxylate functionalized co-monomer to temperature-responsive polymer-grafted cell culture surfaces. Surf Sci. 2004;570:134–41.

    Article  Google Scholar 

  7. Tsuda Y, Kikuchi A, Yamato M, Nakao A, Sakurai Y, Umezu M, Okano T. The use of patterned dual thermoresponsive surfaces for the collective recovery as co-cultured cell sheets. Biomaterials. 2005;26:1885–93.

    Article  Google Scholar 

  8. Schmaljohann D, Oswald J, Jorgensen B, Nitschke M, Beyerlein D, Werner C. Thermo-responsive PNiPAAm-g-PEG films for controlled cell detachment. Biomacromolecules. 2003;4:1733–9.

    Article  Google Scholar 

  9. Nitschke N, Gramm S, Gotze T, Valtink M, Drichel J, Voit B, Engelmann K, Werner C. Thermo-responsive poly(NIPAAm-co-DEGMA) substrates for gentle harvest of human corneal endothelial cell sheets. J Biomed Mater Res A. 2007;80A:1003–10.

    Article  Google Scholar 

  10. Isenberg BC, Tsuda Y, Williams C, Shimizu T, Yamato M, Okano T, Wong JY. A thermoresponsive, microtextured substrate for cell sheet engineering with defined structural organization. Biomaterials. 2008;29:2565–72.

    Article  Google Scholar 

  11. Hatakeyama H, Kikuchi A, Yamato M, Okano T. Patterned bifunctional designs of thermoresponsive surfaces for spatiotemporally controlled cell adhesion, growth and thermally induced detachment. Biomaterials. 2007;28:3632–43.

    Article  Google Scholar 

  12. Mizutani A, Kikuchi A, Yamato M, Kanazawa H, Okano T. Preparation of thermoresponsive polymer brush surfaces and their interactions with cells. Biomaterials. 2008;29:2073–81.

    Article  Google Scholar 

  13. Takahashi H, Matsuzaka N, Nakayama M, Kikuchi A, Yamato M, Okano T. Terminally functionalized thermoresponsive polymer brushes for simultaneously promoting cell adhesion and cell sheet harvest. Biomacromolecules. 2012;13:253–60.

    Article  Google Scholar 

  14. Matsuda N, Shimizu T, Yamato M, Okano T. Tissue engineering based on cell sheet technology. Adv Mater. 2007;19:3089–99.

    Article  Google Scholar 

  15. Hatakeyama H, Kikuchi A, Yamato M, Okano T. Bio-functionalized thermoresponsive interfaces facilitating cell adhesion and proliferation. Biomaterials. 2006;27:5069–78.

    Article  Google Scholar 

  16. Wischerhoff E, Glatzel S, Uhlig K, Lankenau A, Lutz JF, Laschewsky A. Tuning the thickness of polymer brushes grafted from non-linearly growing multilayer assemblies. Langmuir. 2009;25:5949–56.

    Article  Google Scholar 

  17. Dworak A, Utrata-Wesołek A, Szweda D, Kowalczuk A, Trzebicka B, Anioł J, Sieroń AL, Klama-Baryła A, Kawecki M. Poly[tri(ethyleneglycol) ethyl ether methacrylate]-coated surfaces for controlled fibroblasts culturing. ACS Appl Mater Interfaces. 2013;5:2197–207.

    Article  Google Scholar 

  18. Wischerhoff E, Uhlig K, Lankenau A, Borner HG, Laschewsky A, Duschl C, Lutz JF. Controlled cell adhesion on PEG-based switchable surfaces. Angew Chem Int Edit. 2008;47:5666–8.

    Article  Google Scholar 

  19. de la Rosa V. Poly(2-oxazoline)s as materials for biomedical applications. J Mater Sci Mater Med. 2013;. doi:10.1007/s10856-013-5034-y.

    Google Scholar 

  20. Luxenhofer R, Sahay G, Schulz A, Alakhova D, Bronich TK, Jordan R, Kabanov AV. Structure–property relationship in cytotoxicity and cell uptake of poly(2-oxazoline) amphiphiles. J Control Release. 2011;153:73–82.

    Article  Google Scholar 

  21. Kronek J, Kronekova Z, Luston J, Paulovicova E, Paulovicova L, Mendrek B. In vitro and cytotoxicity studies of poly(2-oxazolines). J Mater Sci Mater Med. 2011;22:1725–34.

    Article  Google Scholar 

  22. Luxenhofer R, Han Y, Schultz A, Tong J, He Z, Kabanov AV, Jordan R. Poly(oxazoline)s as polymer therapeutics. Macromol Rapid Commun. 2012;33:1613–31.

    Article  Google Scholar 

  23. Woodle MC, Engbers CM, Zalipsky S. New amphipatic polymer–lipid conjugates forming long-circulating reticuloendothelial system-evading liposomes. Bioconjugate Chem. 1994;5:493–6.

    Article  Google Scholar 

  24. Lee SC, Kim C, Kwon IC, Chung H, Jeong SY. Polymeric micelles of poly(2-ethyl-2-oxazoline)-block-poly(caprolactone) copolymer as a carrier for paclitaxel. J Control Release. 2003;89:437–46.

    Article  Google Scholar 

  25. Zhang N, Pompe T, Amin I, Luxenhofer R, Werner C, Jordan R. Tailored poly(2-oxazoline) polymer brushes to control protein adsorption and cell adhesion. Macromol Biosci. 2012;12:926–36.

    Article  Google Scholar 

  26. Aoi K, Suzuki H, Okada M. Architectural control of sugar-containing polymers by living polymerization: ring-opening polymerization of 2-oxazolines initiated with carbohydrate derivatives. Macromolecules. 1992;25:7073–5.

    Article  Google Scholar 

  27. Guillerm B, Monge S, Lapinte V, Robin JJ. How to modulate the chemical structure of polyoxazolines by appropriate functionalization. Macromol Rapid Commun. 2012;33:1600–12.

    Article  Google Scholar 

  28. Schlaad H, Diehl C, Gress A, Meyer M, Demirel A, Nur Y, Bertin A. Poly(oxazoline)s as smart bioinspired polymers. Macromol Rapid Commun. 2010;31:511–25.

    Article  Google Scholar 

  29. Makino A, Kobayashi S. Chemistry of 2-oxazolines: a crossing of cationic ring-opening polymerization and enzymatic ring-opening addition. J Polym Sci Pol Chem. 2010;48:1251–70.

    Article  Google Scholar 

  30. Dworak A, Trzebicka B, Wałach W. 2-oxazolines, polymerization. In: Salamone JC, editor. Polymeric materials encyclopedia. Boca Raton: CRC Press; 1996. p. 4842–52.

    Google Scholar 

  31. Agrawal M, Rueda JC, Uhlmann P, Muller M, Simon F, Stamm M. Facile approach to grafting of poly(2-oxazoline) brushes on macroscopic surfaces and applications. ACS Appl Mater Interfaces. 2012;4:1357–64.

    Article  Google Scholar 

  32. Lehmann T, Ruhe J. Polyethyloxazoline monolayers for polymer supported biomembrane models. Macromol Symp. 1999;142:1–12.

    Article  Google Scholar 

  33. Jordan R, Ulman A. Surface initiated living cationic polymerization of 2-oxazolines. J Am Chem Soc. 1998;120:243–7.

    Article  Google Scholar 

  34. Jordan J, West N, Ulman A, Chou YM, Nuyken O. Nanocomposites by surface-initiated living cationic polymerization of 2-oxazolines on functionalized gold nanoparticles. Macromolecules. 2001;34:1606–11.

    Article  Google Scholar 

  35. Hutter NA, Reitinger A, Garrido JA, Jordan R. Biofunctionalization of patterned poly(2-oxazoline) bottle-brush brushes on diamond. Polym Preprints. 2012;53:303–4.

    Google Scholar 

  36. Hutter NA, Steenackers M, Reitinger A, Williams OA, Garrido JA, Jordan R. Nanostructured polymer brushes and protein density gradients on diamond by carbon templating. Soft Matter. 2011;7:4861–7.

    Article  Google Scholar 

  37. Zhang N, Steenackers M, Luxenhofer R, Jordan R. Bottle-brush brushes: cylindrical molecular brushes of poly(2-oxazoline) on glassy carbon. Macromolecules. 2009;42:5345–51.

    Article  Google Scholar 

  38. Rehfeldt F, Tanaka M, Pagnoni L, Jordan R. Static and dynamic swelling of grafted poly(2-alkyl-2-oxazoline)s. Langmuir. 2002;18:4908–14.

    Article  Google Scholar 

  39. Loontjens T, Rique-Lurbet L. Synthesis of alkyl trimethoxysilane polyoxazolines and their application as coatings on glass fibres. Des Monomers Polym. 1999;2:217–29.

    Article  Google Scholar 

  40. Wang H, Li L, Tong Q, Yan M. Evaluation of photochemically immobilized poly(2-ethyl-2-oxazoline) thin films as protein-resistant surfaces. ACS Appl Mater Interfaces. 2011;3:3463–71.

    Article  Google Scholar 

  41. Jordan R, Graf K, Riegel H, Unger K. Polymer-supported alkyl monolayers on silica: synthesis and self-assembly of terminal functionalized poly(N-propionylethylenimine)s. Chem Commun. 1996;9:1025–6.

    Article  Google Scholar 

  42. Hutter NA, Reitinger A, Zhang N, Steenackers M, Williams OA, Garrido JA, Jordan R. Microstructured poly(2-oxazoline) bottle-brush brushes on nanocrystalline diamond. Phys Chem Chem Phys. 2010;12:4360–6.

    Article  Google Scholar 

  43. Witte H, Seeliger W. Cyclische imidsaureester aus nitrilern und aminoalkoholen. Liebigs Ann Chem. 1974;6:996–1009.

    Article  Google Scholar 

  44. Utrata-Wesołek A, Oleszko N, Trzebicka B, Anioł J, Zagdańska M, Lesiak M, Sieroń AL, Dworak A. Modified polyglycidol based nanolayers of switchable philicity and their interactions with skin cells. Eur Polym J. 2013;49:106–17.

    Article  Google Scholar 

  45. Hoogenboom R, Fijten MWM, Wijnans S, van den Berg AMJ, Thijs HML, Schubert US. High-throughput synthesis and screening of a library of random and gradient copoly(2-oxazoline)s. J Comb Chem. 2006;8:145–8.

    Article  Google Scholar 

  46. Christova D, Velichkova R, Loos W, Goethals EJ, Du Prez F. New thermo-responsive polymer materials based on poly(2-ethyl-2-oxazoline) segments. Polymer. 2003;44:2255–61.

    Article  Google Scholar 

  47. Howarter JA, Youngblood JP. Optimization of silica silanization by 3-aminopropyltriethoxysilane. Langumir. 2006;22:11142–7.

    Article  Google Scholar 

  48. Meyer M, Antonietti M, Schlaad H. Unexpected thermal characteristics of aqueous solutions of poly(2-isopropyl-2-oxazoline). Soft Matter. 2007;3:430–1.

    Article  Google Scholar 

  49. Demirel AL, Meyer M, Schlaad H. Formation of polyamide nanofibers by directional crystallization in aqueous solution. Angew Chem Int Ed. 2007;46:8622–4.

    Article  Google Scholar 

  50. Morimoto N, Obeid R, Yamane S, Winnik FM, Akiyoshi K. Composite nanomaterials by self-assembly and controlled crystallization of poly(2-isopropyl-2-oxazoline)-grafted polysaccharides. Soft Matter. 2009;5:1597–600.

    Article  Google Scholar 

  51. Diehl C, Dambrowsky I, Hoogenboom R, Schlaad H. Self-assembly of poly(2-alkyl-2-oxazoline)s by crystallization in ethanol–water mixtures below the upper critical solution temperature. Macromol Rapid Commun. 2011;32:1753–8.

    Article  Google Scholar 

  52. Hoeppener S, Wiesbrock F, Hoogenboom R, Thijs HML, Schubert US. Morphologies of spin-coated films of a library of diblock copoly(2-oxazoline)s and their correlation to the corresponding surface energies. Macromol Rapid Commun. 2006;27:405–11.

    Article  Google Scholar 

  53. Hoogenboom R, Fijten MWM, Paulus RM, Thijs HML, Hoeppener S, Kickelbick G, Schubert US. Accelerated pressure synthesis and characterization of 2-oxazoline block copolymers. Polymer. 2006;47:75–84.

    Article  Google Scholar 

  54. Plunkett KN, Zhu X, Moore JS, Leckband DE. PNIPAM chain collapse depends on the molecular weight and grafting density. Langmuir. 2006;22:4259–66.

    Article  Google Scholar 

  55. Kitano H, Kondo T, Suzuki H, Ohno K. Temperature-responsive polymer-brush constructed on a glass substrate by atom transfer radical polymerization. J Colloid Interface Sci. 2010;345:325–31.

    Article  Google Scholar 

  56. Montagne F, Polesel-Maris J, Pugin R, Heinzelmann H. Poly(N-isopropylacrylamide) thin films densley grafted onto gold surface: preparation, characterization, and dynamic AFM study of temperature-induced chain conformational changes. Langmuir. 2009;25:983–91.

    Article  Google Scholar 

  57. Girotto G, Fabbro C, Braghetta P, Vitale P, Volpin D, Bressan GM. Analysis of transcription of the Col6a1 gene in a specific set of tissues suggests a new variant of enhancer region. J Biol Chem. 2000;275:17381–90.

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by the European Union, European Regional Development Fund, project “DERMOSTIM” UDA-POIG.01.03.01-00-088/08. Part of this work was supported by the National Science Centre, the decision DEC-2012/07/N/ST5/00261. N.O. gratefully acknowledges European Social Fund within the project “Scholarships for the development of Silesia” for financial support. The authors thank Dr. Liliana Szyk-Warszynska from the Institute of Catalysis and Surface Chemistry, Polish Academy of Sciences for performing the ellipsometry measurements.

Conflict of interest

The authors declare no competing financial interest.

Author information

Authors and Affiliations

  1. Center of Polymer and Carbon Materials, Polish Academy of Sciences, M. Curie-Sklodowskiej 34, 41-819, Zabrze, Poland

    Andrzej Dworak, Alicja Utrata-Wesołek, Natalia Oleszko, Wojciech Wałach & Barbara Trzebicka

  2. Department of General, Molecular Biology and Genetics, Medical University of Silesia, Medykow 18, 40-752, Katowice, Poland

    Jacek Anioł & Aleksander L. Sieroń

  3. Center for Burn Treatment, Jana Pawla II, 41-100, Siemianowice Slaskie, Poland

    Agnieszka Klama-Baryła & Marek Kawecki

Authors
  1. Andrzej Dworak
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alicja Utrata-Wesołek
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Natalia Oleszko
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wojciech Wałach
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Barbara Trzebicka
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jacek Anioł
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Aleksander L. Sieroń
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Agnieszka Klama-Baryła
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Marek Kawecki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andrzej Dworak.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 240 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dworak, A., Utrata-Wesołek, A., Oleszko, N. et al. Poly(2-substituted-2-oxazoline) surfaces for dermal fibroblasts adhesion and detachment. J Mater Sci: Mater Med 25, 1149–1163 (2014). https://doi.org/10.1007/s10856-013-5135-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10856-013-5135-7

Keywords

Search

Navigation