Assembly of multilayer arrays of viral nanoparticles via biospecific recognition: a quartz crystal microbalance with dissipation monitoring study.

The development of multilayered thin film assemblies containing (bio)molecules is driven by the need to miniaturize sensors, reactors, and biochips. Viral nanoparticles (VNPs) have become popular nanobuilding blocks for material fabrication, and our research has focused on the well-characterized plant virus Cowpea mosaic virus (CPMV). In a previous study, we have reported the construction of multilayer VNP assemblies.

Cell adhesion molecule DM-GRASP presented as nanopatterns to neurons regulates attachment and neurite growth.

Adhesion and neurite formation of neurons and neuroblastoma cells critically depends on the lateral spacing of the cell adhesion molecule DM-GRASP offered as nanostructured substrate.

Propagation of mechanical stress through the actin cytoskeleton toward focal adhesions: model and experiment.

We investigate both theoretically and experimentally how stress is propagated through the actin cytoskeleton of adherent cells and consequentially distributed at sites of focal adhesions (FAs). The actin cytoskeleton is modeled as a two-dimensional cable network with different lattice geometries. Both prestrain, resulting from actomyosin contractility, and central application of external force, lead to finite forces at the FAs that are largely independent of the lattice geometry, but strongly depend on the exact spatial distribution of the FAs.

Protein repellent properties of covalently attached PEG coatings on nanostructured SiO(2)-based interfaces.

In this study, we report the systematic comparison of different poly(ethylene glycol) (PEG) self-assembled monolayers on glass with respect to their protein adsorption and cell adhesion resistance. Combining PEGylation with micellar nanolithography allowed the formation of gold nanoparticle arrays on glass and selective coverage of the free glass area by PEG. The gold nanoparticles serve as anchor points for the attachment of individual proteins and peptides such as the cell-matrix adhesion promoting cyclic RGDfK motif or the kinesin motor protein Eg5.

Molecular engineering of cellular environments: cell adhesion to nano-digital surfaces.

Engineering of the cellular microenvironment has become a valuable means to guide cellular activities such as spreading, motility, differentiation, proliferation, or apoptosis. This chapter summarizes recent approaches to surface patterning such as topography and chemical patterning from the micrometer to the nanometer scale, and illustrates their application to cellular studies. Particular attention is devoted to nanolithography with self-assembled diblock copolymer micelles that are biofunctionalized with peptide ligands-a method that offers unsurpassed spatial resolution for the positioning of signaling molecules over extended surface areas.

Biomimetic models of the actin cytoskeleton.

The cytoskeleton is a complex polymer network that plays an essential role in the functionality of eukaryotic cells. It endows cells with mechanical stability, adaptability, and motility. To identify and understand the mechanisms underlying this large variety of capabilities and to possibly transfer them to engineered networks makes it necessary to have in vitro and in silico model systems of the cytoskeleton.

Syndecan-4-dependent Rac1 regulation determines directional migration in response to the extracellular matrix.

Cell migration in wound healing and disease is critically dependent on integration with the extracellular matrix, but the receptors that couple matrix topography to migratory behavior remain obscure. Using nano-engineered fibronectin surfaces and cell-derived matrices, we identify syndecan-4 as a key signaling receptor determining directional migration. In wild-type fibroblasts, syndecan-4 mediates the matrix-induced protein kinase Calpha (PKCalpha)-dependent activation of Rac1 and localizes Rac1 activity and membrane protrusion to the leading edge of the cell, resulting in persistent migration.

Influence of different ECM mimetic peptide sequences embedded in a nonfouling environment on the specific adhesion of human-skin keratinocytes and fibroblasts on deformable substrates.

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.

Mimicking cellular environments by nanostructured soft interfaces.

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.

Site-specific presentation of single recombinant proteins in defined nanoarrays.

The authors describe the deposition of single biomolecules on substrates at defined spacing by pure self-assembly. The substrate is equipped with an array of 8 nm large gold particles which form the template for biomolecule binding. The authors verified the successful binding of single biomolecules via specific antibody labeling and imaging by fluorescence microscopy.

Early keratinocyte differentiation on micropillar interfaces.

We employed topographical patterning to analyze early keratinocyte differentiation on top of microfabricated pillar arrays. Fibronectin immobilized on pillar "heads" yielded a nucleus-associated granular keratin 1 (K1) pattern in immortalized human gingival keratinocytes (IHGK) at pillar interspaces of 14 mum. Decreasing distances of 11and 8 mum revealed cytoplasmic extension of the early differentiation marker K1 on poly(dimethylsiloxane) (PDMS) pillars.

Cell-assisted assembly of colloidal crystallites.

Many cells ingest foreign particles through a process known as phagocytosis. It now turns out that some cell types organize phagocytosed microparticles into crystalline arrays. Much like the classic crystallization of colloidal particles in a thermal bath, crystallization within the cell is driven by the spatial confinement of mutually repelling particles, in this case by the cell membrane.

Cellular chemomechanics at interfaces: sensing, integration and response.

Living cells are complex entities whose remarkable, emergent capacity to sense, integrate, and respond to environmental cues relies on an intricate series of interactions among the cell's macromolecular components. Defects in mechanosensing, transduction,or responses underlie many diseases such as cancers, immune disorders, cardiac hypertrophy, genetic malformations, and neuropathies. Here, we highlight micro- and nanotechnology-based tools that have been used to study how chemical and mechanical cues modulate the responses of single cells in contact with the extracellular environment.

Cell spreading and focal adhesion dynamics are regulated by spacing of integrin ligands.

Integrin-mediated adhesion is regulated by multiple features of the adhesive surface, including its chemical composition, topography, and physical properties. In this study we investigated integrin lateral clustering, as a mechanism to control integrin functions, by characterizing the effect of nanoscale variations in the spacing between adhesive RGD ligands on cell spreading, migration, and focal adhesion dynamics. For this purpose, we used nanopatterned surfaces, containing RGD-biofunctionalized gold dots, surrounded by passivated gaps.

Tuning the orbital angular momentum in optical vortex beams.

We introduce a method to tune the local orbital angular momentum density in an optical vortex beam without changing its topological charge or geometric intensity distribution. We show that adjusting the relative amplitudes a and b of two interfering collinear vortex beams of equal but opposite helicity provides the smooth variation of the orbital angular momentum density in the resultant vortex beam. Despite the azimuthal intensity modulations that arise from the interference, the local orbital angular momentum remains constant on the vortex annulus and scales with the modulation parameter, c = (a-b)/(a+b).

Three-dimensional modeling of mechanical forces in the extracellular matrix during epithelial lumen formation.

Mechanical interactions between cells and extracellular matrix (ECM) mediate epithelial cyst formation. This work relies on the combination of numerical modeling with live cell imaging, to piece together a novel nonintrusive method for determining three-dimensional (3D) mechanical forces caused by shape changes of a multicellular aggregate at the early stages of epithelial cyst formation. We analyzed the evolution of Madin-Darby canine kidney cells in 3D cultures using time-lapse microscopy, with type I collagen gel forming the ECM.

Lateral spacing of integrin ligands influences cell spreading and focal adhesion assembly.

Cell-extracellular matrix (cell-ECM) interactions mediated by integrin receptors are essential for providing positional and environmental information necessary for many cell functions, such as proliferation, differentiation and survival. In vitro studies on cell adhesion to randomly adsorbed molecules on substrates have been limited to sub-micrometer patches, thus preventing the detailed study of structural arrangement of integrins and their ligands. In this article, we illustrate the role of the distance between integrin ligands, namely the RGD (arginine-glycine-aspartate) sequence present in ECM proteins, in the control of cell adhesion.

Cellular unbinding forces of initial adhesion processes on nanopatterned surfaces probed with magnetic tweezers.

To study the dependence of unbinding forces on the distance of molecularly defined and integrin specific c(-RGDfK-) ligand patches in initial cellular adhesion processes, we developed a magnetic tweezers setup for applying vertical forces of up to 200 pN to rat embryonic fibroblasts. The ligand patch distance is controlled with a hexagonally close packed pattern of biofunctionalized gold nanoparticles prepared by block-copolymer micelle nanolithography. Each gold nanoparticle potentially activates up to one alpha(v)beta(3)-integrin.

Tuning surface energies with nanopatterned substrates.

A novel approach to varying the surface energy of biofunctional substrates has been developed, where surface energies are controlled by utilizing tunable nanopatterned substrates. In this study we functionalized the nanopattern with streptavidin, providing an adhesive interface for biotinylated probes. To obtain the surface energies, we applied the Johnson-Kendall-Roberts model to the adhesion-induced deformation of elastic beads.

Microtubule gliding and cross-linked microtubule networks on micropillar interfaces.

We combined biochemical and topographical patterning to achieve motor-driven microtubule gliding on top of microfabricated pillar arrays with limited and controllable surface interactions of gliding microtubules. Kinesins immobilized on pillar heads pushed microtubules from the top of one micropillar to the next bridging up to 20 mum deep gaps filled with buffer solution. Distances of more than 10 mum were by-passed, and microtubule buckling was occasionally observed.

Geometric organization of the extracellular matrix in the control of integrin-mediated adhesion and cell function in osteoblasts.

Cell-extracellular matrix (ECM) interactions play a central role in tissue architecture and turnover. Particularly, integrin-mediated cell adhesion participates in biochemical and physical signals. The aim of this study is to investigate the importance of ECM organization for alveolar bone osteoblasts adhesion and to determine the effects on cell functions such as collagen and fibronectin production.

High-precision steering of multiple holographic optical traps.

Locating and steering entire ensembles of microscopic objects has become extremely practical with the emergence of holographic optical tweezers. Application of this technology to single molecule experiments requires great accuracy in the spatial positioning of optical traps. This paper calculates the theoretical position resolution of a single holographic beam, predicting that sub-nanometer resolution is easily achieved.

Symmetry dependence of holograms for optical trapping.

No iterative algorithm is necessary to calculate holograms for most holographic optical trapping patterns. Instead, holograms may be produced by a simple extension of the prisms-and-lenses method. This formulaic approach yields the same diffraction efficiency as iterative algorithms for any asymmetric or symmetric but nonperiodic pattern of points while requiring less calculation time.