Aki Kallonen

On the difficulty of increasing dental complexity

One of the fascinating aspects of the history of life is the apparent increase in morphological complexity through time, a well known example being mammalian cheek tooth evolution. In contrast, experimental studies of development more readily show a decrease in complexity, again well exemplified by mammalian teeth, in which tooth crown features called cusps are frequently lost in mutant and transgenic mice. Here we report that mouse tooth complexity can be increased substantially by adjusting multiple signalling pathways simultaneously. We cultured teeth in vitro and adjusted ectodysplasin (EDA), activin A and sonic hedgehog (SHH) pathways, all of which are individually required for normal tooth development. We quantified tooth complexity using the number of cusps and a topographic measure of surface complexity. The results show that whereas activation of EDA and activin A signalling, and inhibition of SHH signalling, individually cause subtle to moderate increases in complexity, cusp number is doubled when all three pathways are adjusted in unison. Furthermore, the increase in cusp number does not result from an increase in tooth size, but from an altered primary patterning phase of development. The combination of a lack of complex mutants, the paucity of natural variants with complex phenotypes, and our results of greatly increased dental complexity using multiple pathways, suggests that an increase may be inherently different from a decrease in phenotypic complexity.

Analysis of short fibres orientation in steel fibre-reinforced concrete (SFRC) by X-ray tomography

The mechanical properties of fibre composite materials are largely determined by the orientation of fibres within the matrix. Which orientation distribution short fibres follow in different parts of a structural element is still a subject for research and discussions in the scientific community. In this article, we present a modern and advanced method for measuring the orientation of short fibres in steel fibre-reinforced concrete (SFRC) by X-ray microtomography. With this method, a voxel image of the fibres is obtained directly in 3D, and the orientation of each individual fibre is calculated based on a skeletonized representation of this image. Scans of 12 SFRC samples, taken from the central height region of real-size floor slabs, reveal the fibres to be mostly horizontally oriented near the centre of a floor slab and more vertically oriented near the edge; here the alignment with the formwork dominates. The fibre orientation distributions are characterized by several orientation parameters as quantitative measures for the alignment. On the practical side, this method has the potential to be incorporated into the development and production process of SFRC structures to verify how the fibres contribute to capacity.

Changes in Submicrometer Structure of Enzymatically Hydrolyzed Microcrystalline Cellulose

To understand the limitations occurring during enzymatic hydrolysis of cellulosic materials in renewable energy production, we used wide-angle X-ray scattering (WAXS), small-angle X-ray scattering (SAXS), X-ray microtomography, and transmission electron microscopy (TEM) to characterize submicrometer changes in the structure of microcrystalline cellulose (Avicel) digested with the Trichoderma reesei enzyme system. The microtomography measurements showed a clear decrease in particle size in scale of tens of micrometers. In all the TEM pictures, similar elongated and partly ramified structures were observed, independent of the hydrolysis time. The SAXS results of rewetted samples suggested a slight change in the structure in scale of 10-20 nm, whereas the WAXS results confirmed that the degree of crystallinity and the crystal sizes remained unchanged. This indicates that the enzymes act on the surface of cellulose bundles and are unable to penetrate into the nanopores of wet cellulose.

Replaying evolutionary transitions from the dental fossil record

The evolutionary relationships of extinct species are ascertained primarily through the analysis of morphological characters. Character inter-dependencies can have a substantial effect on evolutionary interpretations, but the developmental underpinnings of character inter-dependence remain obscure because experiments frequently do not provide detailed resolution of morphological characters. Here we show experimentally and computationally how gradual modification of development differentially affects characters in the mouse dentition. We found that intermediate phenotypes could be produced by gradually adding ectodysplasin A (EDA) protein in culture to tooth explants carrying a null mutation in the tooth-patterning gene Eda. By identifying development-based character inter-dependencies, we show how to predict morphological patterns of teeth among mammalian species. Finally, in vivo inhibition of sonic hedgehog signalling in Eda null teeth enabled us to reproduce characters deep in the rodent ancestry. Taken together, evolutionarily informative transitions can be experimentally reproduced, thereby providing development-based expectations for character-state transitions used in evolutionary studies.

Tablet preformulations of indomethacin-loaded mesoporous silicon microparticles.

In this study, indomethacin-loaded thermally oxidized mesoporous silicon microparticles (TOPSi-IMC) were formulated into tablets with excipients in order to improve the dissolution and permeability properties of the poorly soluble drug. Formulations of TOPSi-IMC particles and excipients were prepared at different TOPSi-IMC particle ratios (25, 30 and 35%). The formulations were compressed by direct compression technique with a single punch tablet machine. For comparison, a formulation containing the bulk IMC (indomethacin) and the same excipients without thermally oxidized mesoporous silicon microparticles particles (TOPSi) was prepared and compressed into tablets. The TOPSi-IMC tablets were characterised according to weight, thickness, crushing strength, disintegration time and dissolution rate. The results of this study show that TOPSi-IMC particles can be compressed to a conventional tablet. The release rate of the drug and its permeation across intestinal cells model (Caco-2) from TOPSi-IMC tablets was improved compared to the bulk IMC tablets. The dissolution rate and permeability of IMC from the tablets decreased with increasing ratio of the TOPSi-IMC particles in the formulation. The phenomenon is, presumably, a result of the loss of unique pore structure of the particles due to deformation of the particles under the compression load.

The origin and loss of periodic patterning in the turtle shell.

The origin of the turtle shell over 200 million years ago greatly modified the amniote body plan, and the morphological plasticity of the shell has promoted the adaptive radiation of turtles. The shell, comprising a dorsal carapace and a ventral plastron, is a layered structure formed by basal endochondral axial skeletal elements (ribs, vertebrae) and plates of bone, which are overlain by keratinous ectodermal scutes. Studies of turtle development have mostly focused on the bones of the shell; however, the genetic regulation of the epidermal scutes has not been investigated. Here, we show that scutes develop from an array of patterned placodes and that these placodes are absent from a soft-shelled turtle in which scutes were lost secondarily. Experimentally inhibiting Shh, Bmp or Fgf signaling results in the disruption of the placodal pattern. Finally, a computational model is used to show how two coupled reaction-diffusion systems reproduce both natural and abnormal variation in turtle scutes. Taken together, these placodal signaling centers are likely to represent developmental modules that are responsible for the evolution of scutes in turtles, and the regulation of these centers has allowed for the diversification of the turtle shell.

X-ray scattering and microtomography study on the structural changes of never-dried silver birch, European aspen and hybrid aspen during drying

Abstract The impact of drying on the structure of the never-dried hardwood cell wall was studied at nanometer level by means of wide- and small-angle X-ray scattering (WAXS, SAXS), and at micrometer level by X-ray microtomography (μCT). Never-dried silver birch, European aspen and hybrid aspen samples were measured by WAXS in situ during drying in air. The samples included juvenile and mature wood, as well as normal and tension wood to allow comparison of the effects of different matrix compositions and microfibril angles. The deformations of cellulose crystallites and amorphous components of the cell wall were detected as changes in the cellulose reflections 200 and 004 and amorphous halo in the WAXS patterns. Especially, the width of the reflection 004, corresponding to the cellulose chain direction, increased due to drying in all the samples, indicating an increase of strain and disorder of the chains. Also, the cellulose unit cell shrank 0.2–0.3% during drying in this direction in all the samples except in hybrid aspen tension wood. According to the SAXS results of silver birch, the distance between micro-fibrils decreased during drying. It was detected by μCT that the mean cross-sectional maximum width of the parenchymatous rays decreased from that of never-dried to air-dried birch by roughly 16%.

Structure of oak wood from the Swedish warship Vasa revealed by X-ray scattering and microtomography

Abstract The degradation of oak wood of the historical warship Vasa was studied, focusing on cellular structure by X-ray microtomography (μCT) and on the nanostructure of the cell wall by wide- and small-angle X-ray scattering (WAXS, SAXS). Solid samples [polyethylene glycol (PEG)-, impregnated and PEG-extracted] were submitted to X-ray analysis and the results compared to those of recent oak. The cellular structure of the Vasa oak was surprisingly well preserved at the micrometer level, according to the μCT images. As revealed by WAXS, the fraction of crystalline cellulose was lower in the Vasa samples compared with recent oak, but the average length and width of cellulose crystallites (25±2 nm and 3.0±0.1 nm, respectively), and the mean microfibril angles (4–9°), showed no significant differences. Accordingly, the crystalline parts of cellulose microfibrils are well preserved in the Vasa oak. The SAXS results indicated a declined short-range order between the cellulose microfibrils and a higher porosity of the Vasa oak compared with recent oak, which may be explained by modification of the hemicellulose-lignin matrix.

Mechanical modelling of tooth wear.

Different diets wear teeth in different ways and generate distinguishable wear and microwear patterns that have long been the basis of palaeodiet reconstructions. Little experimental research has been performed to study them together. Here, we show that an artificial mechanical masticator, a chewing machine, occluding real horse teeth in continuous simulated chewing (of 100 000 chewing cycles) is capable of replicating microscopic wear features and gross wear on teeth that resemble wear in specimens collected from nature. Simulating pure attrition (chewing without food) and four plant material diets of different abrasives content (at n = 5 tooth pairs per group), we detected differences in microscopic wear features by stereomicroscopy of the chewing surface in the number and quality of pits and scratches that were not always as expected. Using computed tomography scanning in one tooth per diet, absolute wear was quantified as the mean height change after the simulated chewing. Absolute wear increased with diet abrasiveness, originating from phytoliths and grit. In combination, our findings highlight that differences in actual dental tissue loss can occur at similar microwear patterns, cautioning against a direct transformation of microwear results into predictions about diet or tooth wear rate.

High-resolution X-ray computed microtomography: A holistic approach to metamorphic fabric analyses

An intrinsic limitation of studying microstructures in thin section is that their spatial (three-dimensional, 3-D) distribution, shape, and orientation have to be inferred by combining 2-D data from different sections. This procedure always involves some degree of interpretation that in some cases can be ambiguous. Recent advances in high-resolution X-ray computed microtomography have made possible the direct imaging in 3-D of volumes of rock to centimeter scale. This rapidly evolving technology is nondestructive and provides a holistic approach of microstructural analysis that eliminates interpretative procedures associated with 2-D methods. Spatial images can be generated through any part of the rock sample and used as virtual petrographic sections. Our application of this technique to an oriented drill core sample from the classic Orijarvi metamorphic region of southern Finland reveals a number of in situ 3-D aspects, including: (1) the spatial distribution and shape of andalusite porphyroblasts, (2) the geometry of a matrix foliation anastomosing around the porphyroblasts, (3) a millimeter-scale compositional layering that controlled the oscillation of porphyroblasts and sulfide mineralization, and (4) distinct inclusion trail patterns characterizing porphyroblast core versus rim zones. The combined data indicate that the steeply dipping bedding-subparallel foliation that characterizes the Orijarvi area formed by bulk north-south crustal shortening and associated vertical stretching.