Mathias Göken

Cell‐based resurfacing of large cartilage defects: Long‐term evaluation of grafts from autologous transgene‐activated periosteal cells in a porcine model of osteoarthritis

OBJECTIVE To investigate the potential of transgene-activated periosteal cells for permanently resurfacing large partial-thickness cartilage defects. METHODS In miniature pigs, autologous periosteal cells stimulated ex vivo by bone morphogenetic protein 2 gene transfer, using liposomes or a combination of adeno-associated virus (AAV) and adenovirus (Ad) vectors, were applied on a bioresorbable scaffold to chondral lesions comprising the entire medial half of the patella. The resulting repair tissue was assessed, 6 and 26 weeks after transplantation, by histochemical and immunohistochemical methods. The biomechanical properties of the repair tissue were characterized by nanoindentation measurements. Implants of unstimulated cells and untreated lesions served as controls. RESULTS All grafts showed satisfactory integration into the preexisting cartilage. Six weeks after transplantation, AAV/Ad-stimulated periosteal cells had adopted a chondrocyte-like phenotype in all layers; the newly formed matrix was rich in proteoglycans and type II collagen, and its contact stiffness was close to that of healthy hyaline cartilage. Unstimulated periosteal cells and cells activated by liposomal gene transfer formed only fibrocartilaginous repair tissue with minor contact stiffness. However, within 6 months following transplantation, the AAV/Ad-stimulated cells in the superficial zone tended to dedifferentiate, as indicated by a switch from type II to type I collagen synthesis and reduced contact stiffness. In deeper zones, these cells retained their chondrocytic phenotype, coinciding with positive staining for type II collagen in the matrix. CONCLUSION Large partial-thickness cartilage defects can be resurfaced efficiently with hyaline-like cartilage formed by transgene-activated periosteal cells. The long-term stability of the cartilage seems to depend on physicobiochemical factors that are active only in deeper zones of the cartilaginous tissue.

Strain-rate sensitivity of ultrafine-grained materials

Abstract The strain-rate sensitivity of commercial purity Aluminium (Al 99.5) and of α-iron, with both conventional (CG) and ultrafine (UFG) grain sizes, are investigated by compression and tension tests at different temperatures. Microstructural investigations were performed before and after compression testing in order to investigate the microstructural stability. Pronounced strain rate sensitivity was found for UFG Al at room temperature as well as at elevated temperatures, while for UFG α-iron an enhanced strain-rate sensitivity was found only at elevated temperatures.

Indentation size effect in spherical and pyramidal indentations

The indentation size effect (ISE) is studied for spherical and pyramidal indentations on a Ni poly-crystal. The indentation experiments were conducted using a Berkovich geometry as well as different spherical indenters with radii of 0.38, 3.8 and 51.0??m. A strong ISE is observed for the material yielding a higher hardness at smaller depths or smaller sphere radii. The transition from elastic to plastic behaviour is associated with a pop-in in the load?displacement curve, in contrast to the conventional elastic?plastic transition as discussed by Tabor. The indentation response is modelled using Tabors approach in conjunction with the uniaxial macroscopic stress?strain behaviour for calculating the statistically stored dislocation density for a given indenter geometry. The geometrically necessary dislocation (GND) density is calculated using a modified Nix/Gao approach, whereas the storage volume for GNDs is used as a parameter for the measured depth dependence of hardness. It will be shown that the ISE for both pyramidal and spherical indentations is related and can be understood within the same given framework. The indentation response of metallic materials can thus be modelled from pop-in to macroscopic hardness.

Characterization of phases of aluminized nickel base superalloys

Aluminum rich coatings, built up by a diffusion zone and a NiAl-cover layer, can protect the surface of turbine blades against oxidation. Within the single crystalline substrate and the adjacent layer, phases in the range of several tens of nanometers up to a few micrometers develop during production and operation of the turbine blade, were characterized. Investigations with transmission electron microscopy, nanoindentation and local crystal orientation mapping with a scanning electron microscope have been carried out in order to determine composition, morphology and distribution of the different phases. The diffusion zone has in general a defined orientation relative to the superalloy substrate and is built up by at least three phases embedded in a softer matrix, with significant differences in nanohardness. Local internal stress states in the diffusion zone are estimated. The NiAl-cover layer is a coarse columnar grained, non-textured B2 ordered intermetallic NiAl-phase.

Determination of plastic properties of polycrystalline metallic materials by nanoindentation: Experiments and finite element simulations

Nanoindentation experiments at low indentation depths are strongly influenced by micromechanical effects, such as the indentation size effect, pile-up or sink-in behaviour and crystal orientation of the investigated material. For an evaluation of load–displacement data and a reconstruction of stress–strain curves from nanoindentations, these micromechanical effects need to be considered. The influence of size effects on experiments were estimated by comparing the results of finite element simulation and experiments, using uniaxial stress–strain data of the indented material as input for the simulations. The experiments were performed on conventional and ultrafine-grained copper and brass, and the influence of the indentation size effect and pile-up formation is discussed in terms of microstructure. Applying a pile-up correction on Berkovich and cube-corner indentation data, a piecewise reconstruction of stress–strain curves from load–displacement data is possible with Tabors concept of representative strain. A good approximation of the slope of the stress–strain curve from the indentation experiments is found for all materials down to an indentation depth of 800 nm.

Deformation behaviour, microstructure and processing of accumulative roll bonded aluminium alloy AA6016

Abstract The technologically relevant aluminium alloy AA6016 was successfully processed using accumulative roll bonding to 8 cycles (∊von Mises = 6.4) in order to obtain an ultrafine-grained microstructure with an average grain size of approximately 200 nm. With the aim of optimising the accumulative roll bonding process detailed investigations on the robustness, the type of rolling mill, the influence of temperature, the microstructural evolution and the mechanical properties have been carried out. Processing at 230 °C provided a good compromise between thermal stability and interlamellar bonding. Samples strained up to 6 cycles (∊von Mises = 4.8) showed an increase in the yield strength by a factor of 3 in comparison to the as-received material. The ductility of the roll bonded samples was slightly sacrificed, although an increase in ductility can be achieved by increasing the number of accumulative roll bonding cycles. The mechanical properties depend on the strain rate, as has also been found for many other ultrafine-grained materials. Annealing of ultrafine-grained samples revealed a stability limit of approximately 200 °C.

The mechanical properties of different lamellae and domains in PST-TiAl investigated with nanoindentations and atomic force microscopy

A nanoindenting atomic force microscope (NI-AFM) was used to determine the hardness and modulus of polysynthetically twinned TiAl on a nanometer scale. Samples with different orientation, (110)γ and (111)γ, were examined. The hardness of α2 lamellae is significantly higher than that of γ lamellae, whereas different γ domains only show small hardness variations. Compression deformed PST crystals show a large amount of twinning in some γ domains. The hardness of twinned and untwinned lamellae are compared. The plastic anisotropy in the γ-phase produces significant differences in the pile-up around the impressions left by nanoindentations. The atomic force microscope allows direct imaging of these impressions. Conical tips were used to study the asymmetric pile-up behavior of the impressions. These patterns are characteristic for the orientation of the lamellae and domains.

Microstructure-dependent deformation behaviour of bcc-metals – indentation size effect and strain rate sensitivity

In this work, the indentation size effect and the influence of the microstructure on the time-dependent deformation behaviour of body-centred cubic (bcc) metals are studied by performing nanoindentation strain rate jump tests at room temperature. During these experiments, the strain rate is abruptly changed, and from the resulting hardness difference the local strain rate sensitivity has been derived. Single-crystalline materials exhibit a strong indentation size effect; ultrafine-grained metals have nearly a depth-independent hardness. Tungsten as a bcc metal shows the opposite behaviour as generally found for face-centered cubic metals. While for UFG-W only slightly enhanced strain rate sensitivity was observed, SX-W exhibits a pronounced influence of the strain rate on the resulting hardness at room temperature. This is due to the effects of the high lattice friction of bcc metals at low temperatures, where the thermally activated motion of screw dislocations is the dominating deformation mechanisms, which causes the enhanced strain rate sensitivity. For the SX-materials, it was found that the degree of the indentation size effect directly correlates with the homologous testing temperature and thus, the material specific parameter of the critical temperature Tc. However, for the resultant strain rate sensitivity no depth-dependent change was found.

Micromechanics and ultrastructure of pyrolysed softwood cell walls

Pyrolytic conversion causes severe changes in the microstructure of the wood cell wall. Pine wood pyrolysed up to 325 °C was investigated by transmission electron microscopy, atomic force microscopy and nanoindentation measurements to monitor changes in structure and mechanical properties. Latewood cell walls were tested in the axial, radial and tangential directions at different temperatures of pyrolysis. A strong anisotropy of elastic properties in the native cell wall was found. Loss of the hierarchical structure of the cell wall due to pyrolysis resulted in elastic isotropy at 300 °C. The development of the mechanical properties with increasing temperature can be explained by alterations in the structure and it was found that the elastic properties were clearly related to length and orientation of the microfibrils.

In situ bulge testing in an atomic force microscope : Microdeformation experiments of thin film membranes

A new bulge testing setup for the measurement of the mechanical properties of thin films is presented. This self-built device can be incorporated in an atomic force microscope (AFM), which allows the recording of topographic images of the observed sample membranes under load conditions. Bulge test experiments on different silicon nitride films are presented and compared to nanoindentation experiments. The measured elastic moduli from nanoindentation and bulge testing are in good agreement. Apart from that, the ability to extract stress–strain data from AFM scans is shown, and the results are compared to standard bulge testing experiments. Imaging of the sample microstructure under load conditions is demonstrated on a thin Cu film.