Articular surface interactions distinguish dinosaurian locomotor joint poses.

Our knowledge of vertebrate functional evolution depends on inferences about joint function in extinct taxa. Without rigorous criteria for evaluating joint articulation, however, such analyses risk misleading reconstructions of vertebrate animal motion. Here we propose an approach for synthesizing raycast-based measurements of 3-D articular overlap, symmetry, and congruence into a quantitative "articulation score" for any non-interpenetrating six-degree-of-freedom joint configuration. We apply our methodology to bicondylar hindlimb joints of two extant dinosaurs (guineafowl, emu) and, through comparison with in vivo kinematics, find that locomotor joint poses consistently have high articulation scores. We then exploit this relationship to constrain reconstruction of a pedal walking stride cycle for the extinct dinosaur Deinonychus antirrhopus, demonstrating the utility of our approach. As joint articulation is investigated in more living animals, the framework we establish here can be expanded to accommodate additional joints and clades, facilitating improved understanding of vertebrate animal motion and its evolution.

Technical note: A volumetric method for measuring the longitudinal arch of human tracks and feet.

Fossil footprints (i.e., tracks) were believed to document arch anatomical evolution, although our recent work has shown that track arches record foot kinematics instead. Analyses of track arches can thereby inform the evolution of human locomotion, although quantifying this 3-D aspect of track morphology is difficult. Here, we present a volumetric method for measuring the arches of 3-D models of human tracks and feet, using both Autodesk Maya and Blender software. The method involves generation of a 3-D object that represents the space beneath the longitudinal arch, and measurement of that arch object's geometry and spatial orientation. We provide relevant tools and guidance for users to apply this technique to their own data. We present three case studies to demonstrate potential applications. These include, (1) measuring the arches of static and dynamic human feet, (2) comparing the arches of human tracks with the arches of the feet that made them, and (3) direct comparisons of human track and foot arch morphology throughout simulated track formation. The volumetric measurement tool proved robust for measuring 3-D models of human tracks and feet, in static and dynamic contexts. This tool enables researchers to quantitatively compare arches of fossil hominin tracks, in order to derive biomechanical interpretations from them, and/or offers a different approach for quantifying foot morphology in living humans.

Arched footprints preserve the motions of fossil hominin feet.

The longitudinal arch of the human foot is viewed as a pivotal adaptation for bipedal walking and running. Fossil footprints from Laetoli, Tanzania, and Ileret, Kenya, are believed to provide direct evidence of longitudinally arched feet in hominins from the Pliocene and Pleistocene, respectively. We studied the dynamics of track formation using biplanar X-ray, three-dimensional animation and discrete element particle simulation. Here, we demonstrate that longitudinally arched footprints are false indicators of foot anatomy; instead they are generated through a specific pattern of foot kinematics that is characteristic of human walking. Analyses of fossil hominin tracks from Laetoli show only partial evidence of this walking style, with a similar heel strike but a different pattern of propulsion. The earliest known evidence for fully modern human-like bipedal kinematics comes from the early Pleistocene Ileret tracks, which were presumably made by members of the genus Homo. This result signals important differences in the foot kinematics recorded at Laetoli and Ileret and underscores an emerging picture of locomotor diversity within the hominin clade.

Inner workings of the alligator ankle reveal the mechanistic origins of archosaur locomotor diversity.

Major transformations in the locomotor system of archosaurs (a major clade of reptiles including birds, crocodiles, dinosaurs, and pterosaurs) were accompanied by significant modifications to ankle anatomy. How the evolution of such a complex multi-joint structure is related to shifts in ankle function and locomotor diversity across this clade remains unclear and weakly grounded in extant experimental data. Here, we used X-ray Reconstruction of Moving Morphology to reconstruct skeletal motion and quantify the sources of three-dimensional ankle mobility in the American alligator, a species that retains the ancestral archosaur ankle structure. We then applied the observed relationships between joint excursion and locomotor behaviors to predict ankle function in extinct archosaurs. High-resolution reconstructions of Alligator skeletal movement revealed previously unseen regionalized coordination among joints responsible for overall ankle rotation. Differences in joint contributions between maneuvers and steady walking parallel transitions in mobility inferred from the ankle structure of fossil taxa in lineages with more erect hind limb postures. Key ankle structures related to ankle mobility were identified in the alligator, which permitted the characterization of ancestral archosaur ankle function. Modifications of these structures provide morphological evidence for functional convergence among sublineages of bird-line and crocodylian-line archosaurs. Using the dynamic insight into the internal sources of Alligator ankle mobility and trends among locomotor modes, we trace anatomical shifts and propose a mechanistic hypothesis for the evolution of ankle structure and function across Archosauria.

The developing bird pelvis passes through ancestral dinosaurian conditions.

Living birds (Aves) have bodies substantially modified from the ancestral reptilian condition. The avian pelvis in particular experienced major changes during the transition from early archosaurs to living birds1,2. This stepwise transformation is well documented by an excellent fossil record2-4; however, the ontogenetic alterations that underly it are less well understood. We used embryological imaging techniques to examine the morphogenesis of avian pelvic tissues in three dimensions, allowing direct comparison with the fossil record. Many ancestral dinosaurian features2 (for example, a forward-facing pubis, short ilium and pubic 'boot') are transiently present in the early morphogenesis of birds and arrive at their typical 'avian' form after transitioning through a prenatal developmental sequence that mirrors the phylogenetic sequence of character acquisition. We demonstrate quantitatively that avian pelvic ontogeny parallels the non-avian dinosaur-to-bird transition and provide evidence for phenotypic covariance within the pelvis that is conserved across Archosauria. The presence of ancestral states in avian embryos may stem from this conserved covariant relationship. In sum, our data provide evidence that the avian pelvis, whose early development has been little studied5-7, evolved through terminal addition-a mechanism8-10 whereby new apomorphic states are added to the end of a developmental sequence, resulting in expression8,11 of ancestral character states earlier in that sequence. The phenotypic integration we detected suggests a previously unrecognized mechanism for terminal addition and hints that retention of ancestral states in development is common during evolutionary transitions.

What is Stance Phase On Deformable Substrates?

The stance phase of walking is when forces are applied to the environment to support, propel, and maneuver the body. Unlike solid surfaces, deformable substrates yield under load, allowing the foot to sink to varying degrees. For bipedal birds and their dinosaurian ancestors, a shared response to walking on these substrates has been identified in the looping path the digits follow underground. Because a volume of substrate preserves a 3-D record of stance phase in the form of footprints or tracks, understanding how the bipedal stride cycle relates to this looping motion is critical for building a track-based framework for the study of walking in extinct taxa. Here we used biplanar X-ray imaging to record and analyze 161 stance phases from 81 trials of three Helmeted Guineafowl (Numida meleagris) walking on radiolucent substrates of different consistency (solid, dry granular, firm to semi-liquid muds). Across all substrates, the feet sank to a range of depths up to 78% of hip height. With increasing substrate hydration, the majority of foot motion shifted from above to below ground. Walking kinematics sampled across all stride cycles revealed six sequential gait-based events originating from both feet, conserved throughout the spectrum of substrate consistencies during normal alternating walking. On all substrates that yielded, five sub-phases of gait were drawn out in space and formed a loop of varying shape. We describe the two-footed coordination and weight distribution that likely contributed to the observed looping patterns of an individual foot. Given such complex subsurface foot motion during normal alternating walking and some atypical walking behaviors, we discuss the definition of "stance phase" on deformable substrates. We also discuss implications of the gait-based origins of subsurface looping on the interpretation of locomotory information preserved in fossil dinosaur tracks.

Advances and Challenges in Paleobiological Reconstructions of Joint Mobility.

Paleobiological reconstructions of joint mobility are an essential component of functional analyses of extinct animals. Over the past half-decade, the methods underlying mobility studies have advanced rapidly in three main areas: increasing complexity of virtual joint manipulation, formalizing pose viability criteria, and constructing more rigorous quantitative frameworks. Here we contextualize and review the recent history of this field, and call attention to remaining challenges and potential future directions. Additionally, we make available and describe a set of user-friendly scripts for the animation software Autodesk Maya. In doing so, we aim to make many of the latest approaches for virtual mobility reconstruction more easily accessible to other researchers, encouraging their broader adoption and collaborative improvement.

A proposed standard for quantifying 3-D hindlimb joint poses in living and extinct archosaurs.

The last common ancestor of birds and crocodylians plus all of its descendants (clade Archosauria) dominated terrestrial Mesozoic ecosystems, giving rise to disparate body plans, sizes, and modes of locomotion. As in the fields of vertebrate morphology and paleontology more generally, studies of archosaur skeletal structure have come to depend on tools for acquiring, measuring, and exploring three-dimensional (3-D) digital models. Such models, in turn, form the basis for many analyses of musculoskeletal function. A set of shared conventions for describing 3-D pose (joint or limb configuration) and 3-D kinematics (change in pose through time) is essential for fostering comparison of posture/movement among such varied species, as well as for maximizing communication among scientists. Following researchers in human biomechanics, we propose a standard methodological approach for measuring the relative position and orientation of the major segments of the archosaur pelvis and hindlimb in 3-D. We describe the construction of anatomical and joint coordinate systems using the extant guineafowl and alligator as examples. Our new standards are then applied to three extinct taxa sampled from the wider range of morphological, postural, and kinematic variation that has arisen across >250 million years of archosaur evolution. These proposed conventions, and the founding principles upon which they are based, can also serve as starting points for measuring poses between elements within a hindlimb segment, for establishing coordinate systems in the forelimb and axial skeleton, or for applying our archosaurian system more broadly to different vertebrate clades.

Integration of biplanar X-ray, three-dimensional animation and particle simulation reveals details of human 'track ontogeny'.

The emergence of bipedalism had profound effects on human evolutionary history, but the evolution of locomotor patterns within the hominin clade remains poorly understood. Fossil tracks record in vivo behaviours of extinct hominins, and they offer great potential to reveal locomotor patterns at various times and places across the human fossil record. However, there is no consensus on how to interpret anatomical or biomechanical patterns from tracks due to limited knowledge of the complex foot-substrate interactions through which they are produced. Here, we implement engineering-based methods to understand human track formation with the ultimate goal of unlocking invaluable information on hominin locomotion from fossil tracks. We first developed biplanar X-ray and three-dimensional animation techniques that permit visualization of subsurface foot motion as tracks are produced, and that allow for direct comparisons of foot kinematics to final track morphology. We then applied the discrete element method to accurately simulate the process of human track formation, allowing for direct study of human track ontogeny. This window lets us observe how specific anatomical and/or kinematic variables shape human track morphology, and it offers a new avenue for robust hypothesis testing in order to infer patterns of foot anatomy and motion from fossil hominin tracks.

Virtual and augmented reality: New tools for visualizing, analyzing, and communicating complex morphology.

Virtual and augmented reality (VR/AR) are new technologies with the power to revolutionize the study of morphology. Modern imaging approaches such as computed tomography, laser scanning, and photogrammetry have opened up a new digital world, enabling researchers to share and analyze morphological data electronically and in great detail. Because this digital data exists on a computer screen, however, it can remain difficult to understand and unintuitive to interact with. VR/AR technologies bridge the analog-to-digital divide by presenting 3D data to users in a very similar way to how they would interact with actual anatomy, while also providing a more immersive experience and greater possibilities for exploration. This manuscript describes VR/AR hardware, software, and techniques, and is designed to give practicing morphologists and educators a primer on using these technologies in their research, pedagogy, and communication to a wide variety of audiences. We also include a series of case studies from the presentations and workshop given at the 2019 International Congress of Vertebrate Morphology, and suggest best practices for the use of VR/AR in comparative morphology.

Paleobiological reconstructions of articular function require all six degrees of freedom.

Paleobiologists typically exclude impossible joint poses from reconstructions of extinct animals by estimating the rotational range of motion (ROM) of fossil joints. However, this ubiquitous practice carries the assumption that osteological estimates of ROM consistently overestimate true joint mobility. Because studies founded on ROM-based exclusion have contributed substantially to our understanding of functional and locomotor evolution, it is critical that this assumption be tested. Here, we evaluate whether ROM-based exclusion is, as currently implemented, a reliable strategy. We measured the true mobilities of five intact cadaveric joints using marker-based X-ray Reconstruction of Moving Morphology and compared them to virtual osteological estimates of ROM made allowing (a) only all three rotational, (b) all three rotational and one translational, and (c) all three rotational and all three translational degrees of freedom. We found that allowing combinations of motions in all six degrees of freedom is necessary to ensure that true mobility is always successfully captured. In other words, failing to include joint translations in ROM analyses results in the erroneous exclusion of many joint poses that are possible in life. We therefore suggest that the functional and evolutionary conclusions of existing paleobiological reconstructions may be weakened or even overturned when all six degrees of freedom are considered. We offer an expanded methodological framework for virtual ROM estimation including joint translations and outline recommendations for future ROM-based exclusion studies.

Alligators employ intermetatarsal reconfiguration to modulate plantigrade ground contact.

Feet must mediate substrate interactions across an animal's entire range of limb poses used in life. Metatarsals, the 'bones of the sole', are the dominant pedal skeletal elements for most tetrapods. In plantigrade species that walk on the entirety of their sole, such as living crocodylians, intermetatarsal mobility offers the potential for a continuum of reconfiguration within the foot itself. Alligator hindlimbs are capable of postural extremes from a belly sprawl to a high walk to sharp turns - how does the foot morphology dynamically accommodate these diverse demands? We implemented a hybrid combination of marker-based and markerless X-ray reconstruction of moving morphology (XROMM) to measure 3D metatarsal kinematics in three juvenile American alligators (Alligator mississippiensis) across their locomotor and maneuvering repertoire on a motorized treadmill and flat-surfaced arena. We found that alligators adaptively conformed their metatarsals to the ground, maintaining plantigrade contact throughout a spectrum of limb placements with non-planar feet. Deformation of the metatarsus as a whole occurred through variable abduction (twofold range of spread) and differential metatarsal pitching (45 deg arc of skew). Internally, metatarsals also underwent up to 65 deg of long-axis rotation. Such reorientation, which correlated with skew, was constrained by the overlapping arrangement of the obliquely expanded metatarsal bases. Such a proximally overlapping metatarsal morphology is shared by fossil archosaurs and archosaur relatives. In these extinct taxa, we suggest that intermetatarsal mobility likely played a significant role in maintaining ground contact across plantigrade postural extremes.

A new role for joint mobility in reconstructing vertebrate locomotor evolution.

Reconstructions of movement in extinct animals are critical to our understanding of major transformations in vertebrate locomotor evolution. Estimates of joint range of motion (ROM) have long been used to exclude anatomically impossible joint poses from hypothesized gait cycles. Here we demonstrate how comparative ROM data can be harnessed in a different way to better constrain locomotor reconstructions. As a case study, we measured nearly 600,000 poses from the hindlimb joints of the Helmeted Guineafowl and American alligator, which represent an extant phylogenetic bracket for the archosaurian ancestor and its pseudosuchian (crocodilian line) and ornithodiran (bird line) descendants. We then used joint mobility mapping to search for a consistent relationship between full potential joint mobility and the subset of joint poses used during locomotion. We found that walking and running poses are predictably located within full mobility, revealing additional constraints for reconstructions of extinct archosaurs. The inferential framework that we develop here can be expanded to identify ROM-based constraints for other animals and, in turn, will help to unravel the history of vertebrate locomotor evolution.

A coordinate-system-independent method for comparing joint rotational mobilities.

Three-dimensional studies of range of motion currently plot joint poses in a 'Euler space' whose axes are angles measured in the joint's three rotational degrees of freedom. Researchers then compute the volume of a pose cloud to measure rotational mobility. However, pairs of poses that are equally different from one another in orientation are not always plotted equally far apart in Euler space. This distortion causes a single joint's mobility to change when measured based on different joint coordinate systems and precludes fair comparison among joints. Here, we present two alternative spaces inspired by a 16th century map projection - cosine-corrected and sine-corrected Euler spaces - that allow coordinate-system-independent comparison of joint rotational mobility. When tested with data from a bird hip joint, cosine-corrected Euler space demonstrated a 10-fold reduction in variation among mobilities measured from three joint coordinate systems. This new quantitative framework enables previously intractable, comparative studies of articular function.

It's in the loop: shared sub-surface foot kinematics in birds and other dinosaurs shed light on a new dimension of fossil track diversity.

The feet of ground-dwelling birds retain many features of their dinosaurian ancestry. Experiments with living species offer insights into the complex interplay among anatomy, kinematics and substrate during the formation of Mesozoic footprints. However, a key aspect of the track-making process, sub-surface foot movement, is hindered by substrate opacity. Here, we use biplanar X-rays to image guineafowl walking through radiolucent substrates of different consistency (solid, dry granular, firm to semi-liquid muds). Despite substantial kinematic variation, the foot consistently moves in a looping pattern below ground. As the foot sinks and then withdraws, the claws of the three main toes create entry and exit paths in different locations. Sampling these paths at incremental horizons captures two-dimensional features just as fossil tracks do, allowing depth-based zones to be characterized by the presence and relative position of digit impressions. Examination of deep, penetrative tracks from the Early Jurassic confirms that bipeds had an equivalent looping response to soft substrates approximately 200 Ma. Our integration of extant and extinct evidence demonstrates the influence of substrate properties on sinking depth and sub-surface foot motion, both of which are significant sources of track variation in the fossil record of dinosaurs.

Contrast-enhanced XROMM reveals in vivo soft tissue interactions in the hip of Alligator mississippiensis.

Extant archosaurs exhibit highly divergent articular soft tissue anatomies between avian and crocodilian lineages. However, the general lack of understanding of the dynamic interactions among archosaur joint soft tissues has hampered further inferences about the function and evolution of these joints. Here we use contrast-enhanced computed tomography to generate 3D surface models of the pelvis, femora, and hip joint soft tissues in an extant archosaur, the American alligator. The hip joints were then animated using marker-based X-Ray Reconstruction of Moving Morphology (XROMM) to visualize soft tissue articulation during forward terrestrial locomotion. We found that the anatomical femoral head of the alligator travels beyond the cranial extent of the bony acetabulum and does not act as a central pivot, as has been suggested for some extinct archosaurs. Additionally, the fibrocartilaginous surfaces of the alligator's antitrochanter and femoral neck remain engaged during hip flexion and extension, similar to the articulation between homologous structures in birds. Moreover, the femoral insertion of the ligamentum capitis moves dorsoventrally against the membrane-bound portion of the medial acetabular wall, suggesting that the inner acetabular foramen constrains the excursion of this ligament as it undergoes cyclical stretching during the step cycle. Finally, the articular surface of the femoral cartilage model interpenetrates with those of the acetabular labrum and antitrochanter menisci; we interpret such interpenetration as evidence of compressive deformation of the labrum and of sliding movement of the menisci. Our data illustrate the utility of XROMM for studying in vivo articular soft tissue interactions. These results also allow us to propose functional hypotheses for crocodilian hip joint soft tissues, expanding our knowledge of vertebrate connective tissue biology and the role of joint soft tissues in locomotor behavior.

A biplanar X-ray approach for studying the 3D dynamics of human track formation.

Recent discoveries have made hominin tracks an increasingly prevalent component of the human fossil record, and these data have the capacity to inform long-standing debates regarding the biomechanics of hominin locomotion. However, there is currently no consensus on how to decipher biomechanical variables from hominin tracks. These debates can be linked to our generally limited understanding of the complex interactions between anatomy, motion, and substrate that give rise to track morphology. These interactions are difficult to study because direct visualization of the track formation process is impeded by foot and substrate opacity. To address these obstacles, we developed biplanar X-ray and computer animation methods, derived from X-ray Reconstruction of Moving Morphology (XROMM), to analyze the 3D dynamics of three human subjects' feet as they walked across four substrates (three deformable muds and rigid composite panel). By imaging and reconstructing 3D positions of external markers, we quantified the 3D dynamics at the foot-substrate interface. Foot shape, specifically heel and medial longitudinal arch deformation, was significantly affected by substrate rigidity. In deformable muds, we found that depths measured across tracks did not directly reflect the motions of the corresponding regions of the foot, and that track outlines were not perfectly representative of foot size. These results highlight the complex, dynamic nature of track formation, and the experimental methods presented here offer a promising avenue for developing and refining methods for accurately inferring foot anatomy and gait biomechanics from fossil hominin tracks.

3-D range of motion envelopes reveal interacting degrees of freedom in avian hind limb joints.

Measuring range of motion (ROM) is a valuable technique that can link bone morphology to joint function in both extant and extinct taxa. ROM results are commonly presented as tables or graphs of maxima and minima for each rotational degree of freedom. We investigate the interactions among three degrees of freedom using X-ray reconstruction of moving morphology (XROMM) to measure ROM of the main hind limb joints of Helmeted Guineafowl (Numida meleagris). By plotting each rotation on an axis, we generate three-dimensional ROM volumes or envelopes composed of hundreds of extreme joint positions for the hip, knee, and intertarsal joints. We find that the shapes of ROM volumes can be quite complex, and that the contribution of long-axis rotation is often substantial. Plotting in vivo poses from individual birds executing different behaviors shows varying use of potential rotational combinations within their ROM envelopes. XROMM can provide unprecedented high-resolution data on the spatial relationship of skeletal elements and thereby illuminate/elucidate the complex ways in which soft and hard tissues interact to produce functional joints. In joints with three rotational degrees of freedom, two-dimensional representations of ROM (flexion/extension and abduction/adduction) are incomplete.

Validation of XMALab software for marker-based XROMM.

Marker-based XROMM requires software tools for: (1) correcting fluoroscope distortion; (2) calibrating X-ray cameras; (3) tracking radio-opaque markers; and (4) calculating rigid body motion. In this paper we describe and validate XMALab, a new open-source software package for marker-based XROMM (C++ source and compiled versions on Bitbucket). Most marker-based XROMM studies to date have used XrayProject in MATLAB. XrayProject can produce results with excellent accuracy and precision, but it is somewhat cumbersome to use and requires a MATLAB license. We have designed XMALab to accelerate the XROMM process and to make it more accessible to new users. Features include the four XROMM steps (listed above) in one cohesive user interface, real-time plot windows for detecting errors, and integration with an online data management system, XMAPortal. Accuracy and precision of XMALab when tracking markers in a machined object are ±0.010 and ±0.043 mm, respectively. Mean precision for nine users tracking markers in a tutorial dataset of minipig feeding was ±0.062 mm in XMALab and ±0.14 mm in XrayProject. Reproducibility of 3D point locations across nine users was 10-fold greater in XMALab than in XrayProject, and six degree-of-freedom bone motions calculated with a joint coordinate system were 3- to 6-fold more reproducible in XMALab. XMALab is also suitable for tracking white or black markers in standard light videos with optional checkerboard calibration. We expect XMALab to increase both the quality and quantity of animal motion data available for comparative biomechanics research.