Dieter P. Kern

Cellular Reactions of Osteoblasts to Micron- and Submicron-Scale Porous Structures of Titanium Surfaces

Osteoblast reactions to topographic structures of titanium play a key role in host tissue responses and the final osseointegration. Since it is difficult to fabricate micro- and nano-scale structures on titanium surfaces, little is known about the mechanism whereby the topography of titanium surfaces exerts its effects on cell behavior at the cellular level. In the present study, the titanium surface was structured in micron- and submicron-scale ranges by anodic oxidation in either 0.2 M H3PO4 or 0.03 M calcium glycerophosphate with 0.15 calcium acetate. The average dimensions of pores in the structured surface were about 0.5 and 2 µm in diameter, with roughness averaging at 0.2 and 0.4 µm, respectively. Enhanced attachment of cells (SaOS-2) was shown on micron- and submicron-scale structures. Initial cell reactions to different titanium surfaces, e.g. the development of the actin-containing structures, are determined by the different morphology of the surfaces. It is demonstrated that on either micron- or submicron-structured surfaces, many well-developed filopodia were observed to be primary adhesion structures in cell-substrate interactions, and some of them entered pores using their distinct tips or points along their length for initial attachment. Therefore, porous structures at either micro- or submicrometre scale supply positive guidance cues for anchorage-dependent cells to attach, leading to enhanced cell attachment. In contrast, the cells attached to a smooth titanium surface by focal contacts around their periphery as predominant adhesion structures, since repulsive signals from the environment led to retraction of the filopodia back to the cell bodies. These cells showed well-organized stress fibres, which exert tension across the cell body, resulting in flattened cells.

Detection of volatile organic compounds (VOCs) with polymer-coated cantilevers

Abstract Changes in the resonance frequency of polymer-coated cantilevers due to gas absorption is shown to be a promising detection mechanism for gases. We prepared SiNx cantilevers based on micromachined Si wafers and used polydimethylsiloxane (PDMS) as polymeric prototype coating. We measured the resonance frequency response of cantilevers end-coated with polymer on exposure to different vapor concentrations of n-octane, toluene, and n-butanole and determined high sensitivities with respect to cantilevers without polymer coating. By using the first higher resonance mode, we found with our configuration sensitivities up to −0.0988 Hz/ppm for small concentrations of n-octane. It is demonstrated here how the sensitivity of the chemical sensor can be improved by varying the deposited polymer mass.

Doped silicon single electron transistors with single island characteristics

Uniformly doped single electron transistors nominally consisting of a single island and two silicon tunneling barriers have been fabricated on silicon–on–insulator material. Two operation regimes are found depending upon the gate voltages applied. The structure acts either as a multiple tunnel junction device or as a single electron transistor consisting of a single dot corresponding to the geometrical shape of the device. The multiple tunnel junction behavior is attributed to the formation of additional tunneling barriers, introduced into the structure by the high doping level. We demonstrate that these barriers can be removed by raising the Fermi level via the application of an appropriate gate voltage.

Multicomponent analysis and prediction with a cantilever array based gas sensor

Abstract We present micromechanical cantilever sensor arrays for quantification of individual components in a gas mixture. Using selectively coated cantilevers as mass sensitive transducers, their response to the analyte molecules is obtained by measuring their resonant frequency shift due to the additional mass loading. The responses were used in principal component regression (partial least squares (PLS)) for the quantitative prediction of vapors being analyzed. The results confirm the functionality of our sensor system and motivate for further optimization.

Gold nanocone near-field scanning optical microscopy probes.

Near-field scanning optical microscopy enables the simultaneous topographical and subdiffraction limited optical imaging of surfaces. A process is presented for the implementation of single individually engineered gold cones at the tips of atomic force microscopy cantilevers. These cantilevers act as novel high-performance optical near-field probes. In the fabrication, thin-film metallization, electron beam induced deposition of etch masks, and Ar ion milling are combined. The cone constitutes a well-defined highly efficient optical antenna with a tip radius on the order of 10 nm and an adjustable plasmon resonance frequency. The sharp tip enables high resolution topographical imaging. By controllably varying the cone size, the resonance frequency can be adapted to the application of choice. Structural properties of these sharp-tipped probes are presented together with topographical images recorded with a cone probe. The antenna functionality is demonstrated by gathering the near-field enhanced Raman signature of individual carbon nanotubes with a gold cone scanning probe.

Cell architecture-cell function dependencies on titanium arrays with regular geometry

Knowledge about biocomplexity of cell behavior in dependence on topographical characteristics is of clinical relevance for the development of implant designs in tissue engineering. The aim of this study was to find out cell architecture-cell function dependencies of human MG-63 osteoblasts on titanium (Ti) arrays with regular geometry. We compared cubic pillar structures (SU-8, dimension 3 x 3 x 5 and 5 x 5 x 5 mum) with planar samples. Electrochemical surface characterization revealed a low amount of surface energy (including polar component) for the pillar-structured surfaces, which correlated with a reduced initial cell adhesion and spreading. Confocal microscopy of cells actin cytoskeleton revealed no stress fiber organization instead, the actin was concentrated in a surface geometry-dependent manner as local spots around the pillar edges. This altered cell architecture resulted in an impaired MG-63 cell function - the extracellular matrix proteins collagen-I and bone sialo protein (BSP-2) were synthesized at a significantly lower level on SU-8 pillar structures; this was accompanied by reduced beta3-integrin expression. To find out physicochemical factors pertaining to geometrically microstructured surfaces and their influence on adjoining biosystems is important for the development of biorelevant implant surfaces.

Influence of temperature on HSQ electron-beam lithography

The authors present a study of the influence of temperature on hydrogen silsesquioxane (HSQ) e-beam lithography during drying, developing, and postdevelopment baking. In accordance with the observation that tempering at relatively low temperatures can already lead to noticeable cross-linking, comparable to the effect of e-beam exposure, the authors find that decreasing the prebake temperature below 90°C and drying the HSQ resist at room temperature in vacuum yields better resolution compared with resist that was dried in a furnace or on a hotplate at 90°C or above. Developing the exposed resist not at room temperature (23°C) but at 60°C results in significant contrast enhancement. Further solidification of the developed resist is obtained by baking the material above 300°C. Correlations between these findings and IR data are presented.

Self-limiting and pattern dependent oxidation of silicon dots fabricated on silicon-on-insulator material

We present a systematic investigation of the oxidation properties of Si dots fabricated on a silicon-on-insulator (SOI) wafer. Dots with diameters varying from 9 to 81 nm were structured on a SOI wafer. These dots were oxidized in a dry oxygen atmosphere at 700, 850, and 1000 °C. The resulting structures were investigated using a side view transmission electron microscopy (TEM) technique in combination with energy filtered TEM. The dimensions of the residual Si and the grown SiO2 were then extracted from the micrographs and analyzed. The oxidation appears to be retarded as compared to the well-known planar oxidation. At 700 and 850 °C a self-limiting effect is observed as well as a clear pattern dependent oxidation at 850 and 1000 °C. We attribute these effects to stress buildup in the oxide. The critical stress, causing the self-limiting effect, is calculated using a model that considers the decrease of the reaction rate with increasing stress perpendicular to the Si surface.

Three-dimensional optical antennas: Nanocones in an apertureless scanning near-field microscope

A sharp-tipped gold nanocone and the vertically aligned metallic tip of a near-field optical microscope together form a three-dimensional optical antenna with a highly controllable gap. Confocal measurements with different laser modes show the efficient axial excitation of the cones with a longitudinally polarized field. In the antenna configuration, extremely strong field enhancement up to a factor of 100 is obtained by tuning the gap between the two sharp tips down to few nanometers.

Investigation of cell reactions to microstructured implant surfaces

Abstract Surface topography is one of the key parameters influencing cellular reactions towards artificial materials. Surfaces with defined microstructures may be useful for enhancement of the stable anchorage of transcutaneous implants in connective tissue or for prevention of epithelial downgrowth and subsequent exfoliation. Cell reactions of keratinocytes and fibroblasts were investigated on microstructured titanium experimental surfaces with alternating grooves and ridges in the range between 1–20 μm width and 0.4–2.0 μm depth. While fibroblasts displayed oriented cell growth on the structured surfaces, human keratinocytes failed to show orientation or enhanced number of focal contacts on structures in the 2–10 μm width range. In that respect, an influence of surface structure on initial cell adhesion could not be proven. The influence of a “bioactive” fibronectin (Fn) coating on adhesion and spreading of fibroblasts was tested on smooth and structured titanium model implant surfaces. Cell spreading was enhanced significantly by the fibronectin coating. Under mechanical shear stress conditions which simulated stresses during insertion of dental implants, the stimulating effects of Fn were lost on smooth surfaces due to abrasion of the coating, while complete abrasion was prevented by microstructured surfaces. The combination of microstructures with “bioactive” coatings may be used to trigger specific cell responses in areas of the implant surface with different functionality. Adhesion and growth of different cell types on microstructured surfaces was investigated by a modified technique at the electron microscopy (EM) level. The approach allows the detection of adhesion molecules in the different membrane domains by immunocytochemical gold labelling techniques. Preliminary results with this new technique suggest that vinculin is localized in the grooves rather than on the ridges in our model system.