Osteoprogenitor response to low-adhesion nanotopographies originally fabricated by electron beam lithography

It is considered that cells can use filopodia, or microspikes, to locate sites suitable for adhesion. This has been investigated using a number of mature cell types, but, to our knowledge, not progenitor cells. Chemical and topographical cues on the underlying substrate are a useful tool for producing defined features for cells to respond to. In this study, arrays of nanopits with different symmetries (square or hexagonal arrays with 120 nm diameters, 300 nm center–centre spacings) and osteoprogenitor cells were considered. The pits were fabricated by ultra-high precision electron-beam lithography and then reproduced in polycarbonate by injection moulding with a nickel stamp. Using scanning electron and fluorescence microscopies, the initial interactions of the cells via filopodia have been observed, as have subsequent adhesion and cytoskeletal formation. The results showed increased filopodia interaction with the surrounding nanoarchitecture leading to a decrease in cell spreading, focal adhesion formation and cytoskeletal organisation.

[1]  A S G Curtis,et al.  Investigating the limits of filopodial sensing: a brief report using SEM to image the interaction between 10 nm high nano‐topography and fibroblast filopodia , 2004, Cell biology international.

[2]  Helmut Schift,et al.  Nanoreplication in polymers using hot embossing and injection molding , 2000 .

[3]  A. Koleske.  Do Filopodia Enable the Growth Cone to Find Its Way? , 2003, Science's STKE.

[4]  A S G Curtis,et al.  Polymer-demixed nanotopography: control of fibroblast spreading and proliferation. , 2002, Tissue engineering.

[5]  S. Britland,et al.  Contact guidance of CNS neurites on grooved quartz: influence of groove dimensions, neuronal age and cell type. , 1997, Journal of cell science.

[6]  W. Korohoda,et al.  CONTACT GUIDANCE OF CHICK EMBRYO NEURONS ON SINGLE SCRATCHES IN GLASS AND ON UNDERLYING ALIGNED HUMAN SKIN FIBROBLASTS , 1999, Cell biology international.

[7]  W. Bonfield,et al.  Increasing hydroxyapatite incorporation into poly(methylmethacrylate) cement increases osteoblast adhesion and response. , 2002, Biomaterials.

[8]  W. Bonfield,et al.  In vitro mechanical and biological assessment of hydroxyapatite-reinforced polyethylene composite , 1997, Journal of materials science. Materials in medicine.

[9]  M. Riehle,et al.  Interaction of animal cells with ordered nanotopography. , 2002, IEEE transactions on nanobioscience.

[10]  A Curtis,et al.  Synergistic and hierarchical adhesive and topographic guidance of BHK cells. , 1996, Experimental cell research.

[11]  T. Yanagida,et al.  Single-Molecule Analysis of Chemotactic Signaling in Dictyostelium Cells , 2001, Science.

[12]  A. Curtis,et al.  CONTROL OF CELL BEHAVIOR: TOPOLOGICAL FACTORS. , 1964, Journal of the National Cancer Institute.

[13]  T. Gustafson,et al.  Studies on the cellular basis of morphogenesis in the sea urchin embryo. Directed movements of primary mesenchyme cells in normal and vegetalized larvae. , 1999, Experimental cell research.

[14]  N. Balaban,et al.  Adhesion-dependent cell mechanosensitivity. , 2003, Annual review of cell and developmental biology.

[15]  Paul Martin,et al.  Structures in focus--filopodia. , 2002, The international journal of biochemistry & cell biology.

[16]  K. Shakesheff,et al.  Human osteoprogenitor growth and differentiation on synthetic biodegradable structures after surface modification. , 2001, Bone.

[17]  Nikolaj Gadegaard,et al.  Investigating filopodia sensing using arrays of defined nano-pits down to 35 nm diameter in size. , 2004, The international journal of biochemistry & cell biology.

[18]  C. Wilkinson,et al.  Role of the cytoskeleton in the reaction of fibroblasts to multiple grooved substrata. , 1995, Cell motility and the cytoskeleton.

[19]  P. Devreotes,et al.  Tumor Suppressor PTEN Mediates Sensing of Chemoattractant Gradients , 2002, Cell.

[20]  B. Kasemo,et al.  Material-tissue interfaces: the role of surface properties and processes. , 1994, Environmental health perspectives.

[21]  C. Wilkinson,et al.  Topographical control of cell behaviour. I. Simple step cues. , 1987, Development.

[22]  L. Wolpert,et al.  Isolation of the cell membrane of Amoeba proteus. , 1961, Experimental cell research.

[23]  Christophe Vieu,et al.  Electron beam lithography: resolution limits and applications , 2000 .

[24]  R. Oreffo,et al.  Skeletal progenitor cells and ageing human populations. , 1998, Clinical science.

[25]  C. Wilkinson,et al.  Substratum nanotopography and the adhesion of biological cells. Are symmetry or regularity of nanotopography important? , 2001, Biophysical chemistry.

[26]  B. Kasemo,et al.  Biomaterial and implant surfaces: a surface science approach. , 1988, The International journal of oral & maxillofacial implants.

[27]  S. Affrossman,et al.  Cell response to nano-islands produced by polymer demixing: a brief review. , 2004, IEE proceedings. Nanobiotechnology.

[28]  C. Wilkinson,et al.  Arrays of nano-dots for cellular engineering , 2003 .

[29]  Hywel Morgan,et al.  Superhydrophobicity and superhydrophilicity of regular nanopatterns. , 2005, Nano letters.

[30]  C. McCaig,et al.  Guidance of CNS growth cones by substratum grooves and ridges: effects of inhibitors of the cytoskeleton, calcium channels and signal transduction pathways. , 1997, Journal of cell science.

[31]  A Curtis,et al.  Guidance and activation of murine macrophages by nanometric scale topography. , 1996, Experimental cell research.

[32]  Nikolaj Gadegaard,et al.  Biomimetic Polymer Nanostructures by Injection Molding , 2003 .

[33]  D. Cumming,et al.  Fabrication of 3 nm wires using 100 keV electron beam lithography and poly(methyl methacrylate) resist , 1996 .

[34]  Matthew John Dalby,et al.  Changes in fibroblast morphology in response to nano-columns produced by colloidal lithography. , 2004, Biomaterials.

[35]  K. Burridge,et al.  Focal adhesions, contractility, and signaling. , 1996, Annual review of cell and developmental biology.

[36]  Matthew J Dalby,et al.  Nucleus alignment and cell signaling in fibroblasts: response to a micro-grooved topography. , 2003, Experimental cell research.

[37]  C. Wilkinson,et al.  Topographical control of cell behaviour: II. Multiple grooved substrata. , 1990, Development.

[38]  C. Wilkinson,et al.  Cells react to nanoscale order and symmetry in their surroundings , 2004, IEEE Transactions on NanoBioscience.

[39]  I. Rehman,et al.  Evaluation of in vitro bioactivity and biocompatibility of Bioglass®-reinforced polyethylene composite , 1997, Journal of materials science. Materials in medicine.