Masticatory biomechanics in the rabbit: a multi-body dynamics analysis

Multi-body dynamics is a powerful engineering tool which is becoming increasingly popular for the simulation and analysis of skull biomechanics. This paper presents the first application of multi-body dynamics to analyse the biomechanics of the rabbit skull. A model has been constructed through the combination of manual dissection and three-dimensional imaging techniques (magnetic resonance imaging and micro-computed tomography). Individual muscles are represented with multiple layers, thus more accurately modelling muscle fibres with complex lines of action. Model validity was sought through comparing experimentally measured maximum incisor bite forces with those predicted by the model. Simulations of molar biting highlighted the ability of the masticatory system to alter recruitment of two muscle groups, in order to generate shearing or crushing movements. Molar shearing is capable of processing a food bolus in all three orthogonal directions, whereas molar crushing and incisor biting are predominately directed vertically. Simulations also show that the masticatory system is adapted to process foods through several cycles with low muscle activations, presumably in order to prevent rapidly fatiguing fast fibres during repeated chewing cycles. Our study demonstrates the usefulness of a validated multi-body dynamics model for investigating feeding biomechanics in the rabbit, and shows the potential for complementing and eventually reducing in vivo experiments.     

Finite-element modelling reveals force modulation of jaw adductors in stag beetles

Male stag beetles carry large and heavy mandibles that arose through sexual selection over mating rights. Although the mandibles of Cyclommatus metallifer males are used in pugnacious fights, they are surprisingly slender. Our bite force measurements show a muscle force reduction of 18% for tip biting when compared with bites with the teeth located halfway along the mandibles. This suggests a behavioural adaptation to prevent failure. We confirmed this by constructing finite-element (FE) models that mimic both natural bite situations as well as the hypothetical situation of tip biting without muscle force modulation. These models, based on micro-CT images, investigate the material stresses in the mandibles for different combinations of bite location and muscle force. Young''s modulus of the cuticle was experimentally determined to be 5.1 GPa with the double indentation method, and the model was validated by digital image correlation on living beetles. FE analysis proves to be a valuable tool in the investigation of the trade-offs of (animal) weapon morphology and usage. Furthermore, the demonstrated bite force modulation in male stag beetles suggests the presence of mechanosensors inside the armature.     

Predicting muscle activation patterns from motion and anatomy: modelling the skull of Sphenodon (Diapsida: Rhynchocephalia)

The relationship between skull shape and the forces generated during feeding is currently under widespread scrutiny and increasingly involves the use of computer simulations such as finite element analysis. The computer models used to represent skulls are often based on computed tomography data and thus are structurally accurate; however, correctly representing muscular loading during food reduction remains a major problem. Here, we present a novel approach for predicting the forces and activation patterns of muscles and muscle groups based on their known anatomical orientation (line of action). The work was carried out for the lizard-like reptile Sphenodon (Rhynchocephalia) using a sophisticated computer-based model and multi-body dynamics analysis. The model suggests that specific muscle groups control specific motions, and that during certain times in the bite cycle some muscles are highly active whereas others are inactive. The predictions of muscle activity closely correspond to data previously recorded from live Sphenodon using electromyography. Apparent exceptions can be explained by variations in food resistance, food size, food position and lower jaw motions. This approach shows considerable promise in advancing detailed functional models of food acquisition and reduction, and for use in other musculoskeletal systems where no experimental determination of muscle activity is possible, such as in rare, endangered or extinct species.     

Caecilian jaw-closing mechanics: integrating two muscle systems

Caecilians (Lissamphibia: Gymnophiona) are unique among vertebrates in having two sets of jaw-closing muscles, one on either side of the jaw joint. Using data from high-resolution X-ray radiation computed tomography scans, we modelled the effect of these two muscle groups (mm. levatores mandibulae and m. interhyoideus posterior) on bite force over a range of gape angles, employing a simplified lever arm mechanism that takes into account muscle cross-sectional area and fibre angle. Measurements of lever arm lengths, muscle fibre orientations and physiological cross-sectional area of cranial muscles were available from three caecilian species: Ichthyophis cf. kohtaoensis; Siphonops annulatus; and Typhlonectes natans. The maximal gape of caecilians is restricted by a critical gape angle above which the mm. levatores mandibulae will open the jaw and destabilize the mandibular joint. The presence of destabilizing forces in the caecilian jaw mechanism may be compensated for by a mandibular joint in that the fossa is wrapped around the condyle to resist dislocation. The caecilian skull is streptostylic; the quadrate–squamosal complex moves with respect to the rest of the skull. This increases the leverage of the jaw-closing muscles. We also demonstrate that the unusual jaw joint requires streptostyly because there is a dorsolateral movement of the quadrate–squamosal complex when the jaw closes. The combination of the two jaw-closing systems results in high bite forces over a wide range of gape angles, an important advantage for generalist feeders such as caecilians. The relative sizes and leverage mechanics of the two closing systems allow one to exert more force when the other has a poor mechanical advantage. This effect is seen in all three species we examined. In the aquatic T. natans, with its less well-roofed skull, there is a larger contribution of the mm. levatores mandibulae to total bite force than in the terrestrial I. cf. kohtaoensis and S. annulatus.     

Multiphasic construct studied in an ectopic osteochondral defect model

In vivo osteochondral defect models predominantly consist of small animals, such as rabbits. Although they have an advantage of low cost and manageability, their joints are smaller and more easily healed compared with larger animals or humans. We hypothesized that osteochondral cores from large animals can be implanted subcutaneously in rats to create an ectopic osteochondral defect model for routine and high-throughput screening of multiphasic scaffold designs and/or tissue-engineered constructs (TECs). Bovine osteochondral plugs with 4 mm diameter osteochondral defect were fitted with novel multiphasic osteochondral grafts composed of chondrocyte-seeded alginate gels and osteoblast-seeded polycaprolactone scaffolds, prior to being implanted in rats subcutaneously with bone morphogenic protein-7. After 12 weeks of in vivo implantation, histological and micro-computed tomography analyses demonstrated that TECs are susceptible to mineralization. Additionally, there was limited bone formation in the scaffold. These results suggest that the current model requires optimization to facilitate robust bone regeneration and vascular infiltration into the defect site. Taken together, this study provides a proof-of-concept for a high-throughput osteochondral defect model. With further optimization, the presented hybrid in vivo model may address the growing need for a cost-effective way to screen osteochondral repair strategies before moving to large animal preclinical trials.     

Morphological determinants of bite force capacity in insects: a biomechanical analysis of polymorphic leaf-cutter ants

The extraordinary success of social insects is partially based on division of labour, i.e. individuals exclusively or preferentially perform specific tasks. Task preference may correlate with morphological adaptations so implying task specialization, but the extent of such specialization can be difficult to determine. Here, we demonstrate how the physical foundation of some tasks can be leveraged to quantitatively link morphology and performance. We study the allometry of bite force capacity in Atta vollenweideri leaf-cutter ants, polymorphic insects in which the mechanical processing of plant material is a key aspect of the behavioural portfolio. Through a morphometric analysis of tomographic scans, we show that the bite force capacity of the heaviest colony workers is twice as large as predicted by isometry. This disproportionate ‘boost’ is predominantly achieved through increased investment in muscle volume; geometrical parameters such as mechanical advantage, fibre length or pennation angle are likely constrained by the need to maintain a constant mandibular opening range. We analyse this preference for an increase in size-specific muscle volume and the adaptations in internal and external head anatomy required to accommodate it with simple geometric and physical models, so providing a quantitative understanding of the functional anatomy of the musculoskeletal bite apparatus in insects.     

Multi-layered bird beaks: a finite-element approach towards the role of keratin in stress dissipation

Bird beaks are layered structures, which contain a bony core and an outer keratin layer. The elastic moduli of this bone and keratin were obtained in a previous study. However, the mechanical role and interaction of both materials in stress dissipation during seed crushing remain unknown. In this paper, a multi-layered finite-element (FE) model of the Java finch''s upper beak (Padda oryzivora) is established. Validation measurements are conducted using in vivo bite forces and by comparing the displacements with those obtained by digital speckle pattern interferometry. Next, the Young modulus of bone and keratin in this FE model was optimized in order to obtain the smallest peak von Mises stress in the upper beak. To do so, we created a surrogate model, which also allows us to study the impact of changing material properties of both tissues on the peak stresses. The theoretically best values for both moduli in the Java finch are retrieved and correspond well with previous experimentally obtained values, suggesting that material properties are tuned to the mechanical demands imposed during seed crushing.     

Measurements of the wall shear stress distribution in the outflow tract of an embryonic chicken heart

In order to study the role of blood–tissue interaction in the developing chicken embryo heart, detailed information about the haemodynamic forces is needed. In this study, we present the first in vivo measurements of the three-dimensional distribution of wall shear stress (WSS) in the outflow tract (OFT) of an embryonic chicken heart. The data are obtained in a two-step process: first, the three-dimensional flow fields are measured during the cardiac cycle using scanning microscopic particle image velocimetry; second, the location of the wall and the WSS are determined by post-processing flow velocity data (finding velocity gradients at locations where the flow approaches zero). The results are a three-dimensional reconstruction of the geometry, with a spatial resolution of 15–20 µm, and provides detailed information about the WSS in the OFT. The most significant error is the location of the wall, which results in an estimate of the uncertainty in the WSS values of 20 per cent.     

Transmission and dose�Cresponse experiments for social animals: a reappraisal of the colonization biology of Campylobacter jejuni in chickens

Dose–response experiments characterize the relationship between infectious agents and their hosts. These experiments are routinely used to estimate the minimum effective infectious dose for an infectious agent, which is most commonly characterized by the dose at which 50 per cent of challenged hosts become infected—the ID₅₀. In turn, the ID₅₀ is often used to compare between different agents and quantify the effect of treatment regimes. The statistical analysis of dose–response data typically makes the assumption that hosts within a given dose group are independent. For social animals, in particular avian species, hosts are routinely housed together in groups during experimental studies. For experiments with non-infectious agents, this poses no practical or theoretical problems. However, transmission of infectious agents between co-housed animals will modify the observed dose–response relationship with implications for the estimation of the ID₅₀ and the comparison between different agents and treatments. We derive a simple correction to the likelihood for standard dose–response models that allows us to estimate dose–response and transmission parameters simultaneously. We use this model to show that: transmission between co-housed animals reduces the apparent value of the ID₅₀ and increases the variability between replicates leading to a distinctive all-or-nothing response; in terms of the total number of animals used, individual housing is always the most efficient experimental design for ascertaining dose–response relationships; estimates of transmission from previously published experimental data for Campylobacter spp. in chickens suggest that considerable transmission occurred, greatly increasing the uncertainty in the estimates of dose–response parameters reported in the literature. Furthermore, we demonstrate that accounting for transmission in the analysis of dose–response data for Campylobacter spp. challenges our current understanding of the differing response of chickens with respect to host-age and in vivo passage of bacteria. Our findings suggest that the age-dependence of transmissibility between hosts—rather than their susceptibility to colonization—is the mechanism behind the ‘lag-phase’ reported in commercial flocks, which are typically found to be Campylobacter free for the first 14–21 days of life.     

Cerebrospinal fluid dynamics in the human cranial subarachnoid space: an overlooked mediator of cerebral disease. I. Computational model

Abnormal cerebrospinal fluid (CSF) flow is suspected to be a contributor to the pathogenesis of neurodegenerative diseases such as Alzheimer''s through the accumulation of toxic metabolites, and to the malfunction of intracranial pressure regulation, possibly through disruption of neuroendocrine communication. For the understanding of transport processes involved in either, knowledge of in vivo CSF dynamics is important. We present a three-dimensional, transient, subject-specific computational analysis of CSF flow in the human cranial subarachnoid space (SAS) based on in vivo magnetic resonance imaging. We observed large variations in the spatial distribution of flow velocities with a temporal peak of 5 cm s⁻¹ in the anterior SAS and less than 4 mm s⁻¹ in the superior part. This could reflect dissimilar flushing requirements of brain areas that may show differences in susceptibility to pathological CSF flow. Our methods can be used to compare the transport of metabolites and neuroendocrine substances in healthy and diseased brains.     

More than just slippery: the impact of biofilm on the attachment of non-sessile freshwater mayfly larvae

While terrestrial insects can usually attach directly to a substrate, for aquatic insects the situation is more complicated owing to the presence of a biofilm on the primary substrates. This important fact has been neither the subject of investigation nor commonly taken into account in the interpretation of functional aspects of attachment in mobile freshwater animals. In this study, we investigate the impact of a biofilm on the attachment of living mayfly larvae. We performed in vivo attachment experiments in a flow channel using different substrates with defined surface roughness. Additionally, we measured friction forces directly generated by dissected tarsal claws on the same substrates. On substrates with smooth or slightly rough surfaces, which have little or no surface irregularities large enough for the claws to grasp, the presence of a biofilm significantly increases the friction force of claws. Consequently, larvae can endure higher flow velocities on these smooth substrates. The opposite effect takes place on rough substrates, where the friction force of claws decreases in the presence of a biofilm. Consequently, a biofilm is a critical ecological structure for these larvae, and other aquatic organisms, not only as a food source but also as a factor influencing attachment ability.     

The wake of hovering flight in bats

Hovering means stationary flight at zero net forward speed, which can be achieved by animals through muscle powered flapping flight. Small bats capable of hovering typically do so with a downstroke in an inclined stroke plane, and with an aerodynamically active outer wing during the upstroke. The magnitude and time history of aerodynamic forces should be reflected by vorticity shed into the wake. We thus expect hovering bats to generate a characteristic wake, but this has until now never been studied. Here we trained nectar-feeding bats, Leptonycteris yerbabuenae, to hover at a feeder and using time-resolved stereoscopic particle image velocimetry in conjunction with high-speed kinematic analysis we show that hovering nectar-feeding bats produce a series of bilateral stacked vortex loops. Vortex visualizations suggest that the downstroke produces the majority of the weight support, but that the upstroke contributes positively to the lift production. However, the relative contributions from downstroke and upstroke could not be determined on the basis of the wake, because wake elements from down- and upstroke mix and interact. We also use a modified actuator disc model to estimate lift force, power and flap efficiency. Based on our quantitative wake-induced velocities, the model accounts for weight support well (108%). Estimates of aerodynamic efficiency suggest hovering flight is less efficient than forward flapping flight, while the overall energy conversion efficiency (mechanical power output/metabolic power) was estimated at 13%.     

Cerebrospinal fluid dynamics in the human cranial subarachnoid space: an overlooked mediator of cerebral disease. II. In vitro arachnoid outflow model

The arachnoid membrane (AM) and granulations (AGs) are important in cerebrospinal fluid (CSF) homeostasis, regulating intracranial pressure in health and disease. We offer a functional perspective of the human AM''s transport mechanism to clarify the role of AM in the movement of CSF and metabolites. Using cultures of human AG cells and a specialized perfusion system, we have shown that this in vitro model mimics the in vivo characteristics of unidirectional fluid transport and we present the first report of serum-free permeability values (92.5 µl min⁻¹ mm Hg⁻¹ cm⁻²), which in turn are in agreement with the CSF outflow rates derived from a dynamic, in vivo magnetic resonance imaging-based computational model of the subarachnoid cranial space (130.9 µl min⁻¹ mm Hg⁻¹ cm⁻²). Lucifer yellow permeability experiments have verified the maintenance of tight junctions by the arachnoidal cells with a peak occurring around 21 days post-seeding, which is when all perfusion experiments were conducted. Addition of ruthenium red to the perfusate, and subsequent analysis of its distribution post-perfusion, has verified the passage of perfusate via both paracellular and transcellular mechanisms with intracellular vacuoles of approximately 1 µm in diameter being the predominant transport mechanism. The comparison of the computational and in vitro models is the first report to measure human CSF dynamics functionally and structurally, enabling the development of innovative approaches to modify CSF outflow and will change concepts and management of neurodegenerative diseases resulting from CSF stagnation.     

Relative brain displacement and deformation during constrained mild frontal head impact

This study describes the measurement of fields of relative displacement between the brain and the skull in vivo by tagged magnetic resonance imaging and digital image analysis. Motion of the brain relative to the skull occurs during normal activity, but if the head undergoes high accelerations, the resulting large and rapid deformation of neuronal and axonal tissue can lead to long-term disability or death. Mathematical modelling and computer simulation of acceleration-induced traumatic brain injury promise to illuminate the mechanisms of axonal and neuronal pathology, but numerical studies require knowledge of boundary conditions at the brain–skull interface, material properties and experimental data for validation. The current study provides a dense set of displacement measurements in the human brain during mild frontal skull impact constrained to the sagittal plane. Although head motion is dominated by translation, these data show that the brain rotates relative to the skull. For these mild events, characterized by linear decelerations near 1.5g (g = 9.81 m s⁻²) and angular accelerations of 120–140 rad s⁻², relative brain–skull displacements of 2–3 mm are typical; regions of smaller displacements reflect the tethering effects of brain–skull connections. Strain fields exhibit significant areas with maximal principal strains of 5 per cent or greater. These displacement and strain fields illuminate the skull–brain boundary conditions, and can be used to validate simulations of brain biomechanics.     

In vivo optic nerve head biomechanics: performance testing of a three-dimensional tracking algorithm

Measurement of optic nerve head (ONH) deformations could be useful in the clinical management of glaucoma. Here, we propose a novel three-dimensional tissue-tracking algorithm designed to be used in vivo. We carry out preliminary verification of the algorithm by testing its accuracy and its robustness. An algorithm based on digital volume correlation was developed to extract ONH tissue displacements from two optical coherence tomography (OCT) volumes of the ONH (undeformed and deformed). The algorithm was tested by applying artificial deformations to a baseline OCT scan while manipulating speckle noise, illumination and contrast enhancement. Tissue deformations determined by our algorithm were compared with the known (imposed) values. Errors in displacement magnitude, orientation and strain decreased with signal averaging and were 0.15 µm, 0.15° and 0.0019, respectively (for optimized algorithm parameters). Previous computational work suggests that these errors are acceptable to provide in vivo characterization of ONH biomechanics. Our algorithm is robust to OCT speckle noise as well as to changes in illumination conditions, and increasing signal averaging can produce better results. This algorithm has potential be used to quantify ONH three-dimensional strains in vivo, of benefit in the diagnosis and identification of risk factors in glaucoma.     

Muscle performance in a soft-bodied terrestrial crawler: constitutive modelling of strain-rate dependency.

Experimental data on the passive mechanical properties of the ventral interior lateral muscle of the tobacco hornworm caterpillar, Manduca sexta, are reported. The stress-deformation response of the Manduca muscle is shown to be nonlinear pseudo-elastic, capable of large deformations and subject to stress softening during initial loading cycles. The muscle passive mechanical properties also depend on multiple time-dependent processes. In particular, we show new experimental data from cyclic loading tests of an unstimulated muscle with constant maximum stretch and different, constant engineering strain rates. Then, on the basis of these data a constitutive model is derived to reproduce the main characteristics of this behaviour. In formulating the constitutive model, we consider the muscle as a complex macromolecular structure with fibrous components at numerous size scales. The model uses a phenomenological approach to account for different mechanisms by which passive force changes during applied deformation and how the muscle properties recover after unloading.     

A consistent muscle activation strategy underlies crawling and swimming in Caenorhabditis elegans

Although undulatory swimming is observed in many organisms, the neuromuscular basis for undulatory movement patterns is not well understood. To better understand the basis for the generation of these movement patterns, we studied muscle activity in the nematode Caenorhabditis elegans. Caenorhabditis elegans exhibits a range of locomotion patterns: in low viscosity fluids the undulation has a wavelength longer than the body and propagates rapidly, while in high viscosity fluids or on agar media the undulatory waves are shorter and slower. Theoretical treatment of observed behaviour has suggested a large change in force–posture relationships at different viscosities, but analysis of bend propagation suggests that short-range proprioceptive feedback is used to control and generate body bends. How muscles could be activated in a way consistent with both these results is unclear. We therefore combined automated worm tracking with calcium imaging to determine muscle activation strategy in a variety of external substrates. Remarkably, we observed that across locomotion patterns spanning a threefold change in wavelength, peak muscle activation occurs approximately 45° (1/8th of a cycle) ahead of peak midline curvature. Although the location of peak force is predicted to vary widely, the activation pattern is consistent with required force in a model incorporating putative length- and velocity-dependence of muscle strength. Furthermore, a linear combination of local curvature and velocity can match the pattern of activation. This suggests that proprioception can enable the worm to swim effectively while working within the limitations of muscle biomechanics and neural control.     

Assessing the impact of marine wind farms on birds through movement modelling

Advances in technology and engineering, along with European Union renewable energy targets, have stimulated a rapid growth of the wind power sector. Wind farms contribute to carbon emission reductions, but there is a need to ensure that these structures do not adversely impact the populations that interact with them, particularly birds. We developed movement models based on observed avoidance responses of common eider Somateria mollissima to wind farms to predict, and identify potential measures to reduce, impacts. Flight trajectory data that were  collected post-construction of the Danish Nysted offshore wind farm were used to parameterize competing models of bird movements around turbines. The model most closely fitting the observed data incorporated individual variation in the minimum distance at which birds responded to the turbines. We show how such models can contribute to the spatial planning of wind farms by assessing their extent, turbine spacing and configurations on the probability of birds passing between the turbines. Avian movement models can make new contributions to environmental assessments of wind farm developments, and provide insights into how to reduce impacts that can be identified at the planning stage.