We herein present a novel platform of well-controlled ordered and disordered nanopatterns positioned with a cyclic peptide of arginine-glycine-aspartic acid (RGD) on a bioinert poly(ethylene glycol) background, to study whether the nanoscopic order of spatial patterning of the integrinspecific ligands influences osteoblast adhesion. This is the first time that the nanoscale order of RGD ligand patterns was varied quantitatively, and tested for its impact on the adhesion of tissue cells. Our findings reveal that integrin clustering and such adhesion induced by RGD ligands is dependent on the local order of ligand arrangement on a substrate when the global average ligand spacing is larger than 70 nm; i.e., cell adhesion is "turned off" by RGD nanopattern order and "turned on" by the RGD nanopattern disorder if operating at this range of inter-ligand spacing.Integrin plays a central role in the formation of focal adhesions (FAs), which anchor cells to the extracellular matrix (ECM). 1 High-affinity binding of the integrin transmembrane proteins to ECM ligands has been extensively exploited for tailoring artificial synthetic ECM systems. 2 Thus far, it has been reported that cell responses to the synthetic ECM depend to a large extent on multiple substrate features, such as its chemical composition, 3-6 geometry and topographical features, 7 ligand organization, 8,9 and even substrate stiffness. 10,11 In particular, the spatial organization of the integrin-specific peptide sequence of arginineglycine-aspartic acid (RGD) on non-fouling substrates has attracted much attention. This sequence, contained in many ECM proteins, can be recognized by all five aV integrins (αVβ1, αVβ3, αVβ5, αVβ6, αVβ8), two β1 integrins (α5β1, α8β1) and αIIbβ3. 12 Once ligated, the integrin receptors link the ECM to the cytoskeleton and integrate intracellular and extracellular events. Furthermore, it is known that cellular behaviors such as adhesion, migration, proliferation and differentiation, are quite sensitive to the bioactivity, tether length, interspacing and density of surface RGD ligands in artificial ECM materials. 13-21Recent developments in nanotechnology have given access to the nanoscale organization of RGD ligands in both inorganic and polymeric substrates mimicking ECMs. Research concerning randomly dispersed RGD ligands grafted onto polymeric materials suggested that *Corresponding authors: E-mail: E-mail: Spatz@mf.mpg.de (J.P. Spatz); E-mail: jdding1@fudan.edu.cn (J. Ding). Supporting Information Available: A detailed description of the experimental protocols for sample preparation and characterization is available free of charge via the Internet at http://pubs.acs.org. Nevertheless, there has been no report to date of studies comparing cellular responses to nanostructured surfaces characterized by ordered or disordered organization of biomolecules such as RGD ligands. Herein, we chose to examine this critical issue in cell-nanomaterial interactions by exploring osteoblast adhesion regulated by the nanoscale organ...

We present herein an innovative technique for decorating soft polymer surfaces with metallic nanostructures fabricated by diblock copolymer micelle nanolithography. Thus far, such nanolithography has been limited to plasma-resistant inorganic substrates such as glass. Our new development is based on the transfer of nanopatterns from glass to soft substrates. Special emphasis is given to hydrogel surfaces with respect to their properties for tailoring cell adhesion. Besides planar surfaces, periodic gold nanopatterns on curved surfaces have been fabricated, as demonstrated on the interior surface of a tubelike hydrogel, which potentially mimic situations of vessels in vivo.

Mechanical stress is a decisive factor for the differentiation, proliferation, and general behavior of cells. However, the specific signaling of mechanotransduction is not fully understood. One basic problem is the clear distinction between the different extracellular matrix (ECM) constituents that participate in cellular adhesion and their corresponding signaling pathways. Here, a system is proposed that enables mechanical stimulation of human-skin-derived keratinocytes and human dermal fibroblasts that specifically interact with peptide sequences immobilized on a non-interacting but deformable substrate. The peptide sequences mimic fibronectin, laminin, and collagen type IV, three major components of the ECM. To achieve this, PDMS is activated using ammonia plasma and coated with star-shaped isocyanate-terminated poly(ethylene glycol)-based prepolymers, which results in a functional coating that prevents unspecific cell adhesion. Specific cell adhesion is achieved by functionalization of the layers with the peptide sequences in different combinations. Moreover, a method that enables the decoration of deformable substrates with cell-adhesion peptides in extremely defined nanostructures is presented. The distance and clustering of cell adhesion molecules below 100 nm has been demonstrated to be of utmost importance for cell adhesion. Thus we present a new toolbox that allows for the detailed analysis of the adhesion of human-skin-derived cells on structurally and biochemically decorated deformable substrates.

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Micro-nano patterns of gold on the surface of poly(ethylene glycol) (PEG) hydrogels were prepared. The approach combines the technique of conventional photolithography (a top-down method for micropatterns), block copolymer micelle nanolithography (a bottom-up method for gold nanopatterns), and a linker-assistant technique to transfer a pattern on a hard surface to a polymeric surface. Hybrid micro-nano patterns on hydrogels were characterized using scanning electron microscopy, atomic force microscopy and X-ray photoelectron spectroscopy. The patterned Au nanoparticles were further modified by a peptide containing arginine-glycine-aspatate (RGD). The cell-adhesion contrast of the patterned hydrogel surface was confirmed by preliminary cell experiments.