Linking hunting weaponry to attack strategies in sailfish and striped marlin.

Linking morphological differences in foraging adaptations to prey choice and feeding strategies has provided major evolutionary insights across taxa. Here, we combine behavioural and morphological approaches to explore and compare the role of the rostrum (bill) and micro-teeth in the feeding behaviour of sailfish (Istiophorus platypterus) and striped marlin (Kajikia audax) when attacking schooling sardine prey. Behavioural results from high-speed videos showed that sailfish and striped marlin both regularly made rostrum contact with prey but displayed distinct strategies. Marlin used high-speed dashes, breaking schools apart, often contacting prey incidentally or tapping at isolated prey with their rostra; while sailfish used their rostra more frequently and tended to use a slower, less disruptive approach with more horizontal rostral slashes on cohesive prey schools. Capture success per attack was similar between species, but striped marlin had higher capture rates per minute. The rostra of both species are covered with micro-teeth, and micro-CT imaging showed that species did not differ in average micro-tooth length, but sailfish had a higher density of micro-teeth on the dorsal and ventral sides of their rostra and a higher amount of micro-teeth regrowth, suggesting a greater amount of rostrum use is associated with more investment in micro-teeth. Our analysis shows that the rostra of billfish are used in distinct ways and we discuss our results in the broader context of relationships between morphological and behavioural feeding adaptations across species.

Gaps at the interface between dentine and self-adhesive resin cement in post-endodontic restorations quantified in 3D by phase contrast-enhanced micro-CT.

AIM: To assess the extent of gaps between root dentine and titanium or fibreglass post restorations following cementation with a self-adhesive resin cement. METHODOLOGY: Fourteen root filled maxillary central incisors restored with prefabricated posts made of Fibreglass (n = 7) or Titanium (n = 7) and cemented with RelyX Unicem 2 were imaged by rapid, high-resolution phase contrast-enhanced micro-CT (PCE-CT) in a synchrotron X-ray imaging facility (ID19, ESRF, 34 KeV, 0.65 µm pixel resolution). Reconstructions were used to measure canal, cement and post perimeters and cross-sectional areas and interfacial gaps at 0.1 mm increments in the root canal space, along the cervical region of the tooth. Remnants of endodontic sealer (AH Plus), when present, were also quantified. Mann-Whitney and 2-way ANOVA tests were used to compare findings within slices and between the two post groups. Pearson correlation coefficients (r) were determined between the interfacial gaps and the other measured parameters. RESULTS: Clearly detectable gaps were found in 45% (±14%) of the interfaces between dentine and cement, along the canal in the cervical area of the tooth beneath the core. The length of interfacial gaps was moderately correlated to the canal cross-sectional area, to the canal perimeter and to the canal area filled by cement (R = 0.52 ~ 0.55, P < 0.001). There was no significant difference between samples with fibreglass or titanium (P > 0.01). Both post types had defect-free interfaces with cement. Endodontic sealer remnants were found on ~10% of the canal walls and were moderately correlated to the presence of gaps. Approximately 30% of the sealer-affected interfaces exhibited no detachment between dentine, sealer and cement. CONCLUSIONS: Self-adhesive cements had interfacial gaps along substantial regions of the root canal surface, which was not correlated with the amount of cement in the canal. PCE-CT proved to be an excellent non-destructive method to study root canal restorations of hydrated samples in 3D.

Injury-mediated decrease in locomotor performance increases predation risk in schooling fish.

The costs and benefits of group living often depend on the spatial position of individuals within groups and the ability of individuals to occupy preferred positions. For example, models of predation events for moving prey groups predict higher mortality risk for individuals at the periphery and front of groups. We investigated these predictions in sardine (Sardinella aurita) schools under attack from group hunting sailfish (Istiophorus platypterus) in the open ocean. Sailfish approached sardine schools about equally often from the front and rear, but prior to attack there was a chasing period in which sardines attempted to swim away from the predator. Consequently, all sailfish attacks were directed at the rear and peripheral positions of the school, resulting in higher predation risk for individuals at these positions. During attacks, sailfish slash at sardines with their bill causing prey injury including scale removal and tissue damage. Sardines injured in previous attacks were more often found in the rear half of the school than in the front half. Moreover, injured fish had lower tail-beat frequencies and lagged behind uninjured fish. Injuries inflicted by sailfish bills may, therefore, hinder prey swimming speed and drive spatial sorting in prey schools through passive self-assortment. We found only partial support for the theoretical predictions from current predator-prey models, highlighting the importance of incorporating more realistic predator-prey dynamics into these models.This article is part of the themed issue 'Physiological determinants of social behaviour in animals'.

BMP delivery complements the guiding effect of scaffold architecture without altering bone microstructure in critical-sized long bone defects: A multiscale analysis.

Scaffold architecture guides bone formation. However, in critical-sized long bone defects additional BMP-mediated osteogenic stimulation is needed to form clinically relevant volumes of new bone. The hierarchical structure of bone determines its mechanical properties. Yet, the micro- and nanostructure of BMP-mediated fast-forming bone has not been compared with slower regenerating bone without BMP. We investigated the combined effects of scaffold architecture (physical cue) and BMP stimulation (biological cue) on bone regeneration. It was hypothesized that a structured scaffold directs tissue organization through structural guidance and load transfer, while BMP stimulation accelerates bone formation without altering the microstructure at different length scales. BMP-loaded medical grade polycaprolactone-tricalcium phosphate scaffolds were implanted in 30mm tibial defects in sheep. BMP-mediated bone formation after 3 and 12 months was compared with slower bone formation with a scaffold alone after 12 months. A multiscale analysis based on microcomputed tomography, histology, polarized light microscopy, backscattered electron microscopy, small angle X-ray scattering and nanoindentation was used to characterize bone volume, collagen fiber orientation, mineral particle thickness and orientation, and local mechanical properties. Despite different observed kinetics in bone formation, similar structural properties on a microscopic and sub-micron level seem to emerge in both BMP-treated and scaffold only groups. The guiding effect of the scaffold architecture is illustrated through structural differences in bone across different regions. In the vicinity of the scaffold increased tissue organization is observed at 3 months. Loading along the long bone axis transferred through the scaffold defines bone micro- and nanostructure after 12 months.

Ptychographic X-ray nanotomography quantifies mineral distributions in human dentine.

Bones are bio-composites with biologically tunable mechanical properties, where a polymer matrix of nanofibrillar collagen is reinforced by apatite mineral crystals. Some bones, such as antler, form and change rapidly, while other bone tissues, such as human tooth dentine, develop slowly and maintain constant composition and architecture for entire lifetimes. When studying apatite mineral microarchitecture, mineral distributions or mineralization activity of bone-forming cells, representative samples of tissue are best studied at submicrometre resolution while minimizing sample-preparation damage. Here, we demonstrate the power of ptychographic X-ray tomography to map variations in the mineral content distribution in three dimensions and at the nanometre scale. Using this non-destructive method, we observe nanostructures surrounding hollow tracts that exist in human dentine forming dentinal tubules. We reveal unprecedented quantitative details of the ultrastructure clearly revealing the spatially varying mineralization density. Such information is essential for understanding a variety of natural and therapeutic effects for example in bone tissue healing and ageing.

Unremodeled endochondral bone is a major architectural component of the cortical bone of the rat (Rattus norvegicus).

The laboratory rat is one of the most frequently-used animal models for studying bone biology and skeletal diseases. Here we show that a substantial portion of the cortical bone of mature rats is primary endochondral bone, consisting of a disorganized arrangement of mineralized collagen fibers. We characterize the structure and mechanical properties of the cortical bone of the rat. We show that the cortical bone consists of two architecturally distinct regions. One region, consisting of well-organized circumferential lamellae (CLB), is located in the endosteal and/or the periosteal regions while another, disorganized region, is located in the more central region of the cortex. Unexpectedly, we found that the disorganized region contains many islands of highly mineralized cartilage. Micro tomography showed different structural and compositional properties of the two primary structural elements; the CLB region has lower mineral density, lower porosity, larger but fewer blood vessels and fewer lacunae. However, no difference was found in the average lacunar volume. Additionally the mean indentation modulus of the CLB region was lower than that of the disorganized region. The islands of calcified cartilage were found to be extremely stiff, with an indentation modulus of 33.4 ± 3.5GPa. We conclude that though the cortical bone of rats is in part lamellar, its architecture is markedly different from that of the cortical bone of humans, a fact that must be borne in mind when using the rat as a model animal for studies of human bone biology and disease.

Characterizing the transformation near indents and cracks in clinically used dental yttria-stabilized zirconium oxide constructs.

OBJECTIVES: Tetragonal zirconia polycrystal (TZP) materials are widely used for full ceramic partial dentures, even though their mechanical properties might change during service. A key property for the durability of the constructs is thought to be an inhibition of crack propagation by phase transformation toughening. Because dental prosthesis are ground and polished for adjustment purposes, it is important to understand the effects of mechanical surface treatments, on localized transformation and around the propagating cracks. METHODS: Sintered samples of commercially available 3 mol-% Yittria-doped TZP were ground and polished and the surface structure and phase composition were compared with those of re-transformed annealed samples. Microindentation was used to induce cracks and nanoindentation was performed to determine the local variety of hardness and indentation modulus, coupled with XRD and SEM investigations. RESULTS: Y-TZP polished surfaces exhibited 9% monoclinic phase content and have reduced hardness and indentation modulus amounting 16.3 GPa and 210.6 GPa, respectively. Y-TZP re-transformed annealed sample revealed 19.4 GPa and 242.3 GPa, respectively. A localized reduction of the stiffness around the crack tips on the annealed surface reveals an irregular arrangement of t-m-transformed grains. Electron micrographs show more damage on the transformed surface following microindentation than on indented annealed surfaces. SIGNIFICANCE: Y-TZP prostheses are adapted and roughened by clinicians prior to bonding to teeth. Annealing recovers properties and microstructure that is changed by the adaptation of the outer layer. This might be important to ensure long-term toughening functionality of the dentures and optimal comfort for the patients.

Monoclinic phase transformations of zirconia-based dental prostheses, induced by clinically practised surface manipulations.

Full-ceramic zirconia crowns and bridges have become very popular with dentists and patients because of their excellent esthetics and mechanical properties. We studied phase transformations within the outermost surface layer of 3 mol.% yttria-stabilized zirconia (Y-TZP) samples of small, clinically relevant thicknesses, manipulated by polishing, grinding and fracture as might be encountered in everyday clinical practice. Stress-induced transformations of the tetragonal phase were studied in three dimensions in order to better understand the organization and extent of the monoclinically transformed phase. By means of laboratory- and synchrotron-based X-ray diffraction measurements, coupled with electron microscopy and multimodal tomography, it was possible for the first time to visualize and quantify the phase distributions non-destructively and in three dimensions. Highly variable degrees of local transformation result in ragged transformed zones of very inhomogeneous thickness. The overall thickness of the transformation layers strongly depends on the severity and rate of loading. Gentle diamond cutting resulted in surprisingly low transformation ratios of less than 0.1%. When Y-TZP constructions are manipulated before bonding, toughness of the outer layers is reduced and they may become brittle with important implications for the stability of the bond: dental practitioners thus need to be cautious when altering the surfaces of these materials after sintering.

Identification of root filling interfaces by microscopy and tomography methods.

AIM: To assess differences in observed cross-sectional areas of root canals and filling materials, as imaged by three microscopy and two tomography methods. METHODOLOGY: Six roots filled with laterally compacted Gutta-percha and AH26 were scanned with phase-contrast enhanced microtomography in a synchrotron facility. Reconstructed virtual slices were compared with sections of both wet and acrylic-embedded roots, evaluated also by light and electron microscopy (EM) and laboratory-based microtomography (µCT). The different contrasts of Gutta-percha, voids, sealer and root dentine were identified and correlated. Inner canal border, outer Gutta-percha rim and the external margin of a void were manually delineated, and the enclosed areas were repeatedly measured by three observers. Interobserver and interimaging method differences were tested by 2-way anova with Bonferroni adjustments (P < 0.05). Percentages of Gutta-percha-filled canal areas (PGP) were determined. RESULTS: Phase-contrast enhanced microtomography revealed internal interfaces and detailed 3D volumes of accentuated voids as well as micrometre-sized particles and gaps within the treated roots. Overestimates in the cross-sectional areas were obtained by light microscopy, whereas underestimates were obtained by µCT and EM. Differences exceeded 40%; however, PGP values by all methods were within 5% for the same slice. Differences between observers were sometimes significant, but they were not method related (<3%). CONCLUSIONS: Phase-contrast enhanced microtomography is a powerful non-destructive ex vivo investigation method for studying the interfaces within root canals and filling materials at a micrometre resolution. The method does not require damage-prone sectioning/polishing during sample preparation procedures. Caution should be used when quantifying the extent of Gutta-percha in root fillings by measurements using µCT, light and EM.

Anisotropic Poisson's ratio and compression modulus of cortical bone determined by speckle interferometry.

Young's modulus and Poisson's ratios of 6mm-sized cubes of equine cortical bone were measured in compression using a micro-mechanical loading device. Surface displacements were determined by electronic speckle pattern-correlation interferometry. This method allows for non-destructive testing of very small samples in water. Analyses of standard materials showed that the method is accurate and precise for determining both Young's modulus and Poisson's ratio. Material properties were determined concurrently in three orthogonal anatomic directions (axial, radial and transverse). Young's modulus values were found to be anisotropic and consistent with values of equine cortical bone reported in the literature. Poisson's ratios were also found to be anisotropic, but lower than those previously reported. Poisson's ratios for the radial-transverse and transverse-radial directions were 0.15+/-0.02, for the axial-transverse and axial-radial directions 0.19+/-0.04, and for the transverse-axial and radial-axial direction 0.09+/-0.02 (mean+/-SD). Cubes located only millimetres apart had significantly different elastic properties, showing that significant spatial variation occurs in equine cortical bone.

Fresnel-propagated imaging for the study of human tooth dentin by partially coherent x-ray tomography.

Recent methods of phase imaging in x-ray tomography allow the visualization of features that are not resolved in conventional absorption microtomography. Of these, the relatively simple setup needed to produce Fresnel-propagated tomograms appears to be well suited to probe tooth-dentin where composition as well as microstructure vary in a graded manner. By adapting analytical propagation approximations we provide predictions of the form of the interference patterns in the 3D images, which we compare to numerical simulations as well as data obtained from measurements of water immersed samples. Our observations reveal details of the tubular structure of dentin, and may be evaluated similarly to conventional absorption tomograms. We believe this exemplifies the power of Fresnel-propagated imaging as a form of 3D microscopy, well suited to quantify gradual microstructural-variations in teeth and similar tissues.