Systematic study of osteoblast response to nanotopography by means of nanoparticle-density gradients.

Features over a wide range of length scales affect the biological response to a surface. While the influence of micro-features has been extensively studied, the effect of nano-features has only rarely been systematically investigated. We have developed a simple method to produce nano-featured gradients by kinetically controlled adsorption of negatively charged silica nanoparticles onto positively charged, poly(ethylene imine) (PEI)-coated silicon wafers. Subsequent sintering of the particles allowed a tuning of the particle morphology and resulted in a firm anchoring of the particles to the surface. Particle-density gradients were characterized by atomic force microscopy (AFM). Cell experiments with rat calvarial osteoblasts (RCO) on nano-featured gradients exhibited a significant decrease in proliferation at locations with higher particle coverage. Seven days post seeding, the number of osteoblasts was eight times higher at positions without particles compared to positions with maximum particle coverage. While cells spread well and developed a well-organized actin network in the absence of particles, spreading and formation of a strong actin network was considerably hindered at locations with maximum particle density.

[1]  C. Murphy,et al.  Effects of Substratum Topography on Cell Behavior , 2002 .

[2]  C. Wilkinson,et al.  Applications of nano-patterning to tissue engineering , 2006 .

[3]  S. Tosatti,et al.  Biomimetic modification of titanium dental implant model surfaces using the RGDSP-peptide sequence: a cell morphology study. , 2006, Biomaterials.

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

[5]  Erkki Ruoslahti,et al.  Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule , 1984, Nature.

[6]  A. Curtis,et al.  Tutorial on the biology of nanotopography , 2004, IEEE Transactions on NanoBioscience.

[7]  R. Kane,et al.  Nanometer-Scale Roughness Having Little Effect on the Amount or Structure of Adsorbed Protein , 2003 .

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

[9]  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.

[10]  Horst Kessler,et al.  RGD modified polymers: biomaterials for stimulated cell adhesion and beyond. , 2003, Biomaterials.

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

[12]  D. Brunette,et al.  Mechanical stretching increases the number of cultured bone cells synthesizing DNA and alters their pattern of protein synthesis , 1985, Calcified Tissue International.

[13]  K. Jandt,et al.  Does the nanometre scale topography of titanium influence protein adsorption and cell proliferation? , 2006, Colloids and surfaces. B, Biointerfaces.

[14]  R G Harrison,et al.  ON THE STEREOTROPISM OF EMBRYONIC CELLS. , 1911, Science.

[15]  A Curtis,et al.  Nantotechniques and approaches in biotechnology. , 2001, Trends in biotechnology.

[16]  P Connolly,et al.  Cell guidance by ultrafine topography in vitro. , 1991, Journal of cell science.

[17]  Michael M. Kozlov,et al.  How proteins produce cellular membrane curvature , 2006, Nature Reviews Molecular Cell Biology.

[18]  M. Textor,et al.  Functionalizable Nanomorphology Gradients via Colloidal Self-Assembly. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[19]  Thilo Stehle,et al.  Crystal Structure of the Extracellular Segment of Integrin αVβ3 , 2001, Science.

[20]  Nanofabrication in cellular engineering , 1998 .

[21]  S. Haskill,et al.  Signal transduction from the extracellular matrix , 1993, The Journal of cell biology.

[22]  C J Murphy,et al.  Effects of synthetic micro- and nano-structured surfaces on cell behavior. , 1999, Biomaterials.

[23]  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.

[24]  H. Boyen,et al.  Ordered Deposition of Inorganic Clusters from Micellar Block Copolymer Films , 2000 .

[25]  J. Engel,et al.  Integrin alphaIIb beta3 reconstituted into lipid bilayers is nonclustered in its activated state but clusters after fibrinogen binding. , 1997, Biochemistry.

[26]  T. Webster,et al.  Mechanisms of enhanced osteoblast adhesion on nanophase alumina involve vitronectin. , 2001, Tissue engineering.

[27]  Thilo Stehle,et al.  Crystal Structure of the Extracellular Segment of Integrin αVβ3 in Complex with an Arg-Gly-Asp Ligand , 2002, Science.

[28]  Bengt Herbert Kasemo,et al.  Biological surface science , 1998 .

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

[30]  D. Brunette,et al.  The effects of the surface topography of micromachined titanium substrata on cell behavior in vitro and in vivo. , 1999, Journal of biomechanical engineering.

[31]  U. Acharya,et al.  Reconstitution of vesiculated Golgi membranes into stacks of cisternae: requirement of NSF in stack formation , 1995, The Journal of cell biology.

[32]  Newell R Washburn,et al.  High-throughput investigation of osteoblast response to polymer crystallinity: influence of nanometer-scale roughness on proliferation. , 2004, Biomaterials.

[33]  Joachim P Spatz,et al.  Activation of integrin function by nanopatterned adhesive interfaces. , 2004, Chemphyschem : a European journal of chemical physics and physical chemistry.