Realistic 3D computer model of the gerbil middle ear, featuring accurate morphology of bone and soft tissue structures.

In order to improve realism in middle ear (ME) finite-element modeling (FEM), comprehensive and precise morphological data are needed. To date, micro-scale X-ray computed tomography (µCT) recordings have been used as geometric input data for FEM models of the ME ossicles. Previously, attempts were made to obtain these data on ME soft tissue structures as well. However, due to low X-ray absorption of soft tissue, quality of these images is limited. Another popular approach is using histological sections as data for 3D models, delivering high in-plane resolution for the sections, but the technique is destructive in nature and registration of the sections is difficult. We combine data from high-resolution µCT recordings with data from high-resolution orthogonal-plane fluorescence optical-sectioning microscopy (OPFOS), both obtained on the same gerbil specimen. State-of-the-art µCT delivers high-resolution data on the 3D shape of ossicles and other ME bony structures, while the OPFOS setup generates data of unprecedented quality both on bone and soft tissue ME structures. Each of these techniques is tomographic and non-destructive and delivers sets of automatically aligned virtual sections. The datasets coming from different techniques need to be registered with respect to each other. By combining both datasets, we obtain a complete high-resolution morphological model of all functional components in the gerbil ME. The resulting 3D model can be readily imported in FEM software and is made freely available to the research community. In this paper, we discuss the methods used, present the resulting merged model, and discuss the morphological properties of the soft tissue structures, such as muscles and ligaments.

High-resolution X-ray CT for 3D petrography of ferruginous sandstone for an investigation of building stone decay.

Diestian ferruginous sandstone has been used as the dominant building stone for monuments in the Hageland, a natural landscape in east-central Belgium. Like all rocks, this stone type is sensitive to weathering. Case hardening was observed in combination with blackening of the exterior parts of the dressed stones. To determine the 3D petrography and to identify the structural differences between the exterior and interior parts, X-ray computed tomography was used in combination with more traditional research techniques like optical microscopy and scanning electron microscopy. The 3D characterization of the ferruginous sandstone was performed with a high-resolution X-ray CT scanner (www.ugct.ugent.be) in combination with the flexible 3D analysis software Morpho+, which provides the necessary petrophysical parameters of the scanned samples in 3D. Besides providing the required 3D parameters like porosity, pore-size distribution, grain size, grain orientation, and surface analysis, the results of the 3D analysis can also be visualized, which enables to understand and interpret the analysis results in a straightforward way. The complementarities between high-quality X-ray CT images and flexible 3D software and its relation with the more traditional microscopical research techniques are opening up new gateways in the study of weathering processes of natural building stones.

Refuge from predation, the benefit of living in an extreme acidic environment?

Organisms living in extreme habitats require costly adaptations to cope with these conditions. Among the suggested potential benefits that trade off these costs is refuge from predation. To study these interactions in extreme environments, samples were taken in the cave Cueva de Villa Luz, Tabasco, Mexico, where more than 32 subterranean springs, some H(2)S rich, rise from the floor. Hydrogen sulfide gas plus oxygen is absorbed by freshwater, and oxidation forms concentrated sulfuric acid. Snottites, whitish hollow mucous tubes, hang from the ceiling of the cave. Fluid drops from these snottites were recorded as having pH values of 0-3. We report the discovery of a new species of nematode that thrives in the highly acidic environment of the snottite. Micro CT scan of snottites reveals a complex interaction between the acidic snottite, nematodes, and abundant nematode-eating mites. The nematode adaptation to low pH probably protects them against mite predation, for which nematodes are most likely the most important source of carbon in this sulfur-driven ecosystem.

Mechanical stress, fracture risk and beak evolution in Darwin's ground finches (Geospiza).

Darwin's finches have radiated from a common ancestor into 14 descendent species, each specializing on distinct food resources and evolving divergent beak forms. Beak morphology in the ground finches (Geospiza) has been shown to evolve via natural selection in response to variation in food type, food availability and interspecific competition for food. From a mechanical perspective, however, beak size and shape are only indirectly related to birds' abilities to crack seeds, and beak form is hypothesized to evolve mainly under selection for fracture avoidance. Here, we test the fracture-avoidance hypothesis using finite-element modelling. We find that across species, mechanical loading is similar and approaches reported values of bone strength, thus suggesting pervasive selection on fracture avoidance. Additionally, deep and wide beaks are better suited for dissipating stress than are more elongate beaks when scaled to common sizes and loadings. Our results illustrate that deep and wide beaks in ground finches enable reduction of areas with high stress and peak stress magnitudes, allowing birds to crack hard seeds while limiting the risk of beak failure. These results may explain strong selection on beak depth and width in natural populations of Darwin's finches.

Linking morphology and motion: a test of a four-bar mechanism in seahorses.

Syngnathid fishes (seahorses, pipefish, and sea dragons) possess a highly modified cranium characterized by a long and tubular snout with minute jaws at its end. Previous studies indicated that these species are extremely fast suction feeders with their feeding strike characterized by a rapid elevation of the head accompanied by rotation of the hyoid. A planar four-bar model is proposed to explain the coupled motion of the neurocranium and the hyoid. Because neurocranial elevation as well as hyoid rotation are crucial for the feeding mechanism in previously studied Syngnathidae, a detailed evaluation of this model is needed. In this study, we present kinematic data of the feeding strike in the seahorse Hippocampus reidi. We combined these data with a detailed morphological analysis of the important linkages and joints involved in rotation of the neurocranium and the hyoid, and we compared the kinematic measurements with output of a theoretical four-bar model. The kinematic analysis shows that neurocranial rotation never preceded hyoid rotation, thus indicating that hyoid rotation triggers the explosive feeding strike. Our data suggest that while neurocranium and hyoid initially (first 1.5 ms) behave as predicted by the four-bar model, eventually, the hyoid rotation is underestimated by the model. Shortening, or a posterior displacement of the sternohyoid muscle (of which the posterior end is confluent with the hypaxial muscles in H. reidi), probably explains the discrepancy between the model and our kinematic measurements. As a result, while four-bar modeling indicates a clear coupling between hyoid rotation and neurocranial elevation, the detailed morphological determination of the linkages and joints of this four-bar model remain crucial in order to fully understand this mechanism in seahorse feeding.

Histologic and micro-computed tomographic evaluation of the osseointegration of a nonresorbable bone substitute in alveoli of ponies after tooth extraction.

OBJECTIVE: To evaluate the biological behavior of a nonresorbable bone substitute (NRBS) in the alveoli of ponies, compared with tissue quality in naturally healing alveoli, after cheek tooth extraction. ANIMALS: 5 clinically normal ponies. PROCEDURES: In each pony, both maxillary fourth premolars (Triadan 108/208) were repulsed bilaterally during anesthesia. One randomly chosen alveolus was filled with NRBS and isolated from the oral cavity by use of dental impression material and a spring-wire retention device. The other alveolus was occluded in its occlusal third portion with dental impression material. One year after surgery, cylindrical lateromedial biopsy specimens were collected from the apical, middle, and occlusal level of each alveolus. Biopsy samples were evaluated for bone mineral density and bone volume via micro-computed tomography; qualitative histologic characteristics were evaluated via light microscopy. RESULTS: Bone mineral density and bone volume were greater in control alveoli, compared with NRBS-treated alveoli. Control alveoli were characterized by the presence of few mature bone trabeculae and wide spaces containing fat tissue and mesenchymal stroma. In treated alveoli, biocompatibility and osteoconductive properties of the NRBS were excellent; continuous bone formation and bone remodeling were also evident. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the NRBS was integrated well in calcified alveolar tissues in ponies 1 year after maxillary cheek tooth extraction. Further research is necessary to establish the benefits of this NRBS in the development of a dental implant surgical technique in equids.

Porous gelatin hydrogels: 1. Cryogenic formation and structure analysis.

In the present work, porous gelatin scaffolds were prepared by cryogenic treatment of a chemically cross-linked gelatin hydrogel, followed by removal of the ice crystals formed through lyophilization. This technique often leads to porous gels with a less porous skin. A simple method has been developed to solve this problem. The present study demonstrates that the hydrogel pore size decreased with an increasing gelatin concentration and with an increasing cooling rate of the gelatin hydrogel. Variation of the cryogenic parameters applied also enabled us to develop scaffolds with different pore morphologies (spherical versus transversal channel-like pores). In our opinion, this is the first paper in which temperature gradients during controlled cryogenic treatment were applied to induce a pore size gradient in gelatin hydrogels. With a newly designed cryo-unit, temperature gradients of 10 and 30 degrees C were implemented during the freezing step, resulting in scaffolds with average pore diameters of, respectively, +/-116 and +/-330 microm. In both cases, the porosity and pore size decreased gradually through the scaffolds. Pore size and structure analysis of the matrices was accomplished through a combination of microcomputed tomography using different software packages (microCTanalySIS and Octopus), scanning electron microscopy analysis, and helium pycnometry.

Porous gelatin hydrogels: 2. In vitro cell interaction study.

We report on the feasibility of applying porous gelatin hydrogels, prepared by a novel and controlled cryogenic treatment, as cell-interactive scaffolds for tissue engineering applications. Despite the large number of publications on gelatin as a biomaterial, a detailed study of screening a limited number of gelatin scaffolds for their interaction with a panel of human cells has, to the best of our knowledge, not yet been published. In the present work, we have evaluated two types of porous gelatin scaffolds that differ in their pore geometry and pore size. Type I hydrogels contained top-to-bottom transverse channels (i.e. cones) with a decreasing diameter from the top (330 microm) to the bottom (20-30 microm). Type II hydrogels contained spherical pores with a diameter of 135 microm. Both types of scaffolds were evaluated by confocal laser scanning microscopy in terms of adhesion, spreading, and proliferation of human cells (endothelial, epithelial, fibroblast, glial, and osteoblast) by visualizing cells using calcein-acetoxy methyl ester as a vital stain. The results indicated that cells attached, spread, and proliferated on both types of hydrogels. In addition, the scaffolds developed can be used for the long-term culturing of human cells.

Use of X-ray computed microtomography for non-invasive determination of wood anatomical characteristics.

Quantitative analysis of wood anatomical characteristics is usually performed using classical microtomy yielding optical micrographs of stained thin sections. It is time-consuming to obtain high quality cross-sections from microtomy, and sections can be damaged. This approach, therefore, is often impractical for those who need quick acquisition of quantitative data on vessel characteristics in wood. This paper reports results of a novel approach using X-ray computed microtomography (microCT) for non-invasive determination of wood anatomy. As a case study, stem wood samples of a 2-year-old beech (Fagus sylvatica L.) and a 3-year-old oak (Quercus robur L.) tree were investigated with this technique, beech being a diffuse-porous and oak a ring-porous tree species. MicroCT allowed non-invasive mapping of 2-D transverse cross-sections of both wood samples with micrometer resolution. Self-developed software 'microCTanalysis' was used for image processing of the 2-D cross-sections in order to automatically determine the inner vessel diameters, the transverse cross-sectional surface area of the vessels, the vessel density and the porosity with computer assistance. Performance of this new software was compared with manual analysis of the same micrographs. The automatically obtained results showed no significant statistical differences compared to the manual measurements. Visual inspection of the microCT slices revealed very good correspondence with the optical micrographs. Statistical analysis confirmed this observation in a more quantitative way, and it was, therefore, argued that anatomical analysis of optical micrographs can be readily substituted by automated use of microCT, and this without loss of accuracy. Furthermore, as an additional application of microCT, the 3-D renderings of the internal microstructure of the xylem vessels for both the beech and the oak sample could be reconstructed, clearly showing the complex nature of vessel networks. It can be concluded that the use of microCT in wood science offers an interesting potential for all those who need quantitative data of wood anatomical characteristics in either the 2-D or the 3-D space.