Molecular and Cellular Biochemistry

, Volume 301, Issue 1–2, pp 241–249 | Cite as

Morphology, cytoskeletal organization, and myosin dynamics of mouse embryonic fibroblasts cultured on nanofibrillar surfaces

  • Ijaz Ahmed
  • Abdul S. Ponery
  • Alam Nur-E-Kamal
  • Jabeen Kamal
  • Adam S. Meshel
  • Michael P. Sheetz
  • Melvin Schindler
  • Sally Meiners
Article

Abstract

Growth of cells in tissue culture is generally performed on two-dimensional (2D) surfaces composed of polystyrene or glass. Recent work, however, has shown that such 2D cultures are incomplete and do not adequately represent the physical characteristics of native extracellular matrix (ECM)/basement membrane (BM), namely dimensionality, compliance, fibrillarity, and porosity. In the current study, a three-dimensional (3D) nanofibrillar surface composed of electrospun polyamide nanofibers was utilized to mimic the topology and physical structure of ECM/BM. Additional chemical cues were incorporated into the nanofibrillar matrix by coating the surfaces with fibronectin, collagen I, or laminin-1. Results from the current study show an enhanced response of primary mouse embryonic fibroblasts (MEFs) to culture on nanofibrillar surfaces with more dramatic changes in cell spreading and reorganization of the cytoskeleton than previously observed for established cell lines. In addition, the cells cultured on nanofibrillar and 2D surfaces exhibited differential responses to the specific ECM/BM coatings. The localization and activity of myosin II-B for MEFs cultured on nanofibers was also compared. A dynamic redistribution of myosin II-B was observed within membrane protrusions. This was previously described for cells associated with nanofibers composed of collagen I but not for cells attached to 2D surfaces coated with monomeric collagen. These results provide further evidence that nanofibrillar surfaces offer a significantly different environment for cells than 2D substrates.

Keywords

Nanofibers Nanofibrillar surfaces Extracellular matrix Mouse embryonic fibroblasts Myosin II-B Fibronectin Laminin-1 Collagen I 

References

  1. 1.
    Kim BS, Nikolovski J, Bonadio J, Smiley E, Mooney DJ (1999) Engineered smooth muscle tissues: regulating cell phenotype with the scaffold. Exp Cell Res 251:318–328PubMedCrossRefGoogle Scholar
  2. 2.
    Sakiyama SE, Schense JC, Hubbell JA (1999) Incorporation of heparin-binding peptides into fibrin gels enhances neurite extension: an example of designer matrices in tissue engineering. FASEB J 13:2214–2224PubMedGoogle Scholar
  3. 3.
    Lutolf MP, Lauer-Fields JL, Schmoekel HG, Metters AT, Weber FE, Fields GB, Hubbell JA (2003) Synthetic matrix metalloproteinase-sensitive hydrogels for the conduction of tissue regeneration: engineering cell-invasion characteristics. Proc Natl Acad Sci USA 100:5413–5418PubMedCrossRefGoogle Scholar
  4. 4.
    Schmeichel KL, Bissell MJ (2003) Modeling tissue-specific signaling and organ function in three dimensions. J Cell Sci 116:2377–2388PubMedCrossRefGoogle Scholar
  5. 5.
    Meiners S, Mercado ML (2003) Functional peptide sequences derived from extracellular matrix glycoproteins and their receptors: strategies to improve neuronal regeneration. Mol Neurobiol 27:177–196PubMedCrossRefGoogle Scholar
  6. 6.
    Shin H, Jo S, Mikos AG (2003) Biomimetic materials for tissue engineering. Biomaterials 24:4353–4364PubMedCrossRefGoogle Scholar
  7. 7.
    Cukierman E, Pankov R, Stevens DR, Yamada KM (2001) Taking cell-matrix adhesions to the third dimension. Science 294:1708–1712PubMedCrossRefGoogle Scholar
  8. 8.
    Grinnell F, Ho CH, Tamariz E, Lee DJ, Skuta G (2003) Dendritic fibroblasts in three-dimensional collagen matrices. Mol Biol Cell 14:384–395PubMedCrossRefGoogle Scholar
  9. 9.
    Cukierman E, Pankov R, Yamada KM (2002) Cell interactions with three-dimensional matrices. Curr Opin Cell Biol 14:633–639PubMedCrossRefGoogle Scholar
  10. 10.
    Wozniak MA, Desai R, Solski PA, Der CJ, Keely PJ (2003) ROCK-generated contractility regulates breast epithelial cell differentiation in response to the physical properties of a three-dimensional collagen matrix. J Cell Biol 163:583–595PubMedCrossRefGoogle Scholar
  11. 11.
    Schindler M, Ahmed I, Nur-E-Kamal A, Kamal J, Grafe TH, Chung HY, Meiners S (2005) Synthetic nanofibrillar matrix promotes in vivo-like organization and morphogenesis for cells in culture. Biomaterials 26:5624–5631PubMedCrossRefGoogle Scholar
  12. 12.
    Chung HY, Hal JRB, Gogins MA, Crofoot DG, Weik TM (2004) Polymer, polymer microfiber, polymer nanofiber and applications including filter structures. US Patent No 6,743,273 B2Google Scholar
  13. 13.
    Li WJ, Danielson KG, Alexander PG, Tuan RS (2003) Biological response of chondrocytes cultured in three-dimensional nanofibrous poly(epsilon-caprolactone) scaffolds. J Biomed Mater Res 67A:1105–1114CrossRefGoogle Scholar
  14. 14.
    Yoshimoto H, Shin YM, Terai H, Vacanti JP (2003) A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering. Biomaterials 12:2077–2082CrossRefGoogle Scholar
  15. 15.
    Boland ED, Matthews JA, Pawlowski KJ, Simpson DG, Wnek GE, Bowlin GL. Electrospinning collagen, elastin (2004) preliminary vascular tissue engineering. Front Biosci 9:1422–1432PubMedCrossRefGoogle Scholar
  16. 16.
    Nur-E-Kamal A, Ahmed I, Kamal J, Schindler M, Meiners S (2005) Three dimensional nanofibrillar surfaces induce activation of Rac. Biochem Biophys Res Com 331:428–434PubMedCrossRefGoogle Scholar
  17. 17.
    Ashkenas J, Muschler J, Bissell MJ (1996) The extracellular matrix in epithelial biology: shared molecules and common themes in distant phyla. Dev Biol 180:433–444PubMedCrossRefGoogle Scholar
  18. 18.
    Kalluri R. (2003) Basement membranes: structure, assembly and role in tumour angiogenesis. Nat Rev Cancer 3:422–433PubMedCrossRefGoogle Scholar
  19. 19.
    Kleinman HK, Philp D, Hoffman MP (2003) Role of the extracellular matrix in morphogenesis. Curr Op Biotech 14:526–532CrossRefGoogle Scholar
  20. 20.
    Katz BZ, Zamir E, Bershadsky A, Kam Z, Yamada KM, Geiger B (2000) Physical state of the extracellular matrix regulates the structure and molecular composition of cell-matrix adhesions. Mol Biol Cell 11:1047–1060PubMedGoogle Scholar
  21. 21.
    Hynes RO (1999) The dynamic dialogue between cells and matrices: implications of fibronectin’s elasticity. Proc Natl Acad Sci USA 96:2588–2590PubMedCrossRefGoogle Scholar
  22. 22.
    Meshel AS, Wei Q, Adelstein R, Sheetz MP (2005) Basic mechanism of three-dimensional collagen fibre transport by fibroblasts. Nat Cell Biol 7:157–164PubMedCrossRefGoogle Scholar
  23. 23.
    Geiger B, Bershadsky A, Pankov R, Yamada KM (2001) Transmembrane cross-talk between the extracellular matrix-cytoskeleton. Nature Rev Mol Cell Biol 2:793–805CrossRefGoogle Scholar
  24. 24.
    Zamir E, Geiger B (2001) Molecular complexity and dynamics of cell-matrix adhesions. J Cell Sci 14:3583–3590Google Scholar
  25. 25.
    Craig SW, Chen H (2003) Lamellipodia protrusion: moving interations of vinculin and Arp2/3. Curr Biol 13:R236–238PubMedCrossRefGoogle Scholar
  26. 26.
    Djinović-Carugo K, Young P, Gautel M, Saraste M (1999) Structure of the α-actinin rod: molecular basis for cross-linking of actin filaments. Cell 98:537–546PubMedCrossRefGoogle Scholar
  27. 27.
    Watanabe N, Kato T, Fujita A, Ishizaki T, Narumiya S (1999) Cooperation between mDia1 and ROCK in rho-induced actin reorganization. Nature Cell Biol 1:136–143PubMedCrossRefGoogle Scholar
  28. 28.
    Elliott JT, Woodward JT, Langenbach KJ, Tona A, Jones PL, Plant AL (2005) Vascular smooth muscle cell response on thin films of collagen. Matrix Biol 7:489–502CrossRefGoogle Scholar
  29. 29.
    Beningo KA, Dembo M, Wang Y-L (2004) Responses of fibroblasts to anchorage of dorsal extracellular matrix receptors. Proc Natl Acad Sci USA 101:18024–18029PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Ijaz Ahmed
    • 1
  • Abdul S. Ponery
    • 1
  • Alam Nur-E-Kamal
    • 2
  • Jabeen Kamal
    • 1
  • Adam S. Meshel
    • 3
  • Michael P. Sheetz
    • 3
  • Melvin Schindler
    • 4
  • Sally Meiners
    • 1
  1. 1.Department of PharmacologyUMDNJ-Robert Wood Johnson Medical SchoolPiscatawayUSA
  2. 2.Department of BiologyMedgar-Evers College of the City University of New YorkBrooklynUSA
  3. 3.Department of Biological SciencesColumbia UniversityNew YorkUSA
  4. 4.NanoCulture, LLCPiscatawayUSA

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