Static versus dynamic muscle modelling in extinct species: a biomechanical case study of the Australopithecus afarensis pelvis and lower extremity.

The force a muscle generates is dependent on muscle structure, in which fibre length, pennation angle and tendon slack length all influence force production. Muscles are not preserved in the fossil record and these parameters must be estimated when constructing a musculoskeletal model. Here, we test the capability of digitally reconstructed muscles of the Australopithecus afarensis model (specimen AL 288-1) to maintain an upright, single-support limb posture. Our aim was to ascertain the influence that different architectural estimation methods have on muscle specialisation and on the subsequent inferences that can be extrapolated about limb function. Parameters were estimated for 36 muscles in the pelvis and lower limb and seven different musculoskeletal models of AL 288-1 were produced. These parameters represented either a 'static' Hill-type muscle model (n = 4 variants) which only incorporated force, or instead a 'dynamic' Hill-type muscle model with an elastic tendon and fibres that could vary force-length-velocity properties (n = 3 variants). Each muscle's fibre length, pennation angle, tendon slack length and maximal isometric force were calculated based upon different input variables. Static (inverse) simulations were computed in which the vertical and mediolateral ground reaction forces (GRF) were incrementally increased until limb collapse (simulation failure). All AL 288-1 variants produced somewhat similar simulated muscle activation patterns, but the maximum vertical GRF that could be exerted on a single limb was not consistent between models. Three of the four static-muscle models were unable to support >1.8 times body weight and produced models that under-performed. The dynamic-muscle models were stronger. Comparative results with a human model imply that similar muscle group activations between species are needed to sustain single-limb support at maximally applied GRFs in terms of the simplified static simulations (e.g., same walking pose) used here. This approach demonstrated the range of outputs that can be generated for a model of an extinct individual. Despite mostly comparable outputs, the models diverged mostly in terms of strength.

Three-dimensional volumetric muscle reconstruction of the Australopithecus afarensis pelvis and limb, with estimations of limb leverage.

To understand how an extinct species may have moved, we first need to reconstruct the missing soft tissues of the skeleton, which rarely preserve, with an understanding of segmental volume and muscular composition within the body. The Australopithecus afarensis specimen AL 288-1 is one of the most complete hominin skeletons. Despite 40+ years of research, the frequency and efficiency of bipedal movement in this specimen is still debated. Here, 36 muscles of the pelvis and lower limb were reconstructed using three-dimensional polygonal modelling, guided by imaging scan data and muscle scarring. Reconstructed muscle masses and configurations guided musculoskeletal modelling of the lower limb in comparison with a modern human. Results show that the moment arms of both species were comparable, hinting towards similar limb functionality. Moving forward, the polygonal muscle modelling approach has demonstrated promise for reconstructing the soft tissues of hominins and providing information on muscle configuration and space filling. This method demonstrates that volumetric reconstructions are required to know where space must be occupied by muscles and thus where lines of action might not be feasible due to interference with another muscle. This approach is effective for reconstructing muscle volumes in extinct hominins for which musculature is unknown.

Modern three-dimensional digital methods for studying locomotor biomechanics in tetrapods.

Here, we review the modern interface of three-dimensional (3D) empirical (e.g. motion capture) and theoretical (e.g. modelling and simulation) approaches to the study of terrestrial locomotion using appendages in tetrapod vertebrates. These tools span a spectrum from more empirical approaches such as XROMM, to potentially more intermediate approaches such as finite element analysis, to more theoretical approaches such as dynamic musculoskeletal simulations or conceptual models. These methods have much in common beyond the importance of 3D digital technologies, and are powerfully synergistic when integrated, opening a wide range of hypotheses that can be tested. We discuss the pitfalls and challenges of these 3D methods, leading to consideration of the problems and potential in their current and future usage. The tools (hardware and software) and approaches (e.g. methods for using hardware and software) in the 3D analysis of tetrapod locomotion have matured to the point where now we can use this integration to answer questions we could never have tackled 20 years ago, and apply insights gleaned from them to other fields.

Quantitative biomechanical assessment of locomotor capabilities of the stem archosaur Euparkeria capensis.

Birds and crocodylians are the only remaining members of Archosauria (ruling reptiles) and they exhibit major differences in posture and gait, which are polar opposites in terms of locomotor strategies. Their broader lineages (Avemetatarsalia and Pseudosuchia) evolved a multitude of locomotor modes in the Triassic and Jurassic periods, including several occurrences of bipedalism. The exact timings and frequencies of bipedal origins within archosaurs, and thus their ancestral capabilities, are contentious. It is often suggested that archosaurs ancestrally exhibited some form of bipedalism. Euparkeria capensis is a central taxon for the investigation of locomotion in archosaurs due to its phylogenetic position and intermediate skeletal morphology, and is argued to be representative of facultative bipedalism in this group. However, no studies to date have biomechanically tested if bipedality was feasible in Eupakeria. Here, we use musculoskeletal models and static simulations in its hindlimb to test the influences of body posture and muscle parameter estimation methods on locomotor potential. Our analyses show that the resulting negative pitching moments around the centre of mass were prohibitive to sustainable bipedality. We conclude that it is unlikely that Euparkeria was facultatively bipedal, and was probably quadrupedal, rendering the inference of ancestral bipedal abilities in Archosauria unlikely.

Anatomically grounded estimation of hindlimb muscle sizes in Archosauria.

In vertebrates, active movement is driven by muscle forces acting on bones, either directly or through tendinous insertions. There has been much debate over how muscle size and force are reflected by the muscular attachment areas (AAs). Here we investigate the relationship between the physiological cross-sectional area (PCSA), a proxy for the force production of the muscle, and the AA of hindlimb muscles in Nile crocodiles and five bird species. The limbs were held in a fixed position whilst blunt dissection was carried out to isolate the individual muscles. AAs were digitised using a point digitiser, before the muscle was removed from the bone. Muscles were then further dissected and fibre architecture was measured, and PCSA calculated. The raw measures, as well as the ratio of PCSA to AA, were studied and compared for intra-observer error as well as intra- and interspecies differences. We found large variations in the ratio between AAs and PCSA both within and across species, but muscle fascicle lengths are conserved within individual species, whether this was Nile crocodiles or tinamou. Whilst a discriminant analysis was able to separate crocodylian and avian muscle data, the ratios for AA to cross-sectional area for all species and most muscles can be represented by a single equation. The remaining muscles have specific equations to represent their scaling, but equations often have a relatively high success at predicting the ratio of muscle AA to PCSA. We then digitised the muscle AAs of Coelophysis bauri, a dinosaur, to estimate the PCSAs and therefore maximal isometric muscle forces. The results are somewhat consistent with other methods for estimating force production, and suggest that, at least for some archosaurian muscles, that it is possible to use muscle AA to estimate muscle sizes. This method is complementary to other methods such as digital volumetric modelling.

Reconstructing Articular Cartilage in the Australopithecus afarensis Hip Joint and the Need for Modeling Six Degrees of Freedom.

The postcranial skeleton of Australopithecus afarensis (AL 288-1) exhibits clear adaptations for bipedality, although there is some debate as to the efficiency and frequency of such upright movement. Some researchers argue that AL 288-1 walked with an erect limb like modern humans do, whilst others advocate for a "bent-hip bent-knee" (BHBK) gait, although in recent years the general consensus favors erect bipedalism. To date, no quantitative method has addressed the articulation of the AL 288-1 hip joint, nor its range of motion (ROM) with consideration for joint spacing, used as a proxy for the thickness of the articular cartilage present within the joint spacing which can affect how a joint moves. Here, we employed ROM mapping methods to estimate the joint spacing of AL 288-1's hip joint in comparison to a modern human and chimpanzee. Nine simulations assessed different joint spacing and tested the range of joint congruency (i.e., ranging from a closely packed socket to loosely packed). We further evaluated the sphericity of the femoral head and whether three rotational degrees of freedom (DOFs) sufficiently captures the full ROM or if translational DOFs must be included. With both setups, we found that the AL 288-1 hip was unlikely to be highly congruent (as it is in modern humans) because this would severely restrict hip rotational movement and would severely limit the capability for both bipedality and even arboreal locomotion. Rather, the hip was more cartilaginous than it is in the modern humans, permitting the hip to rotate into positions necessitated by both terrestrial and arboreal movements. Rotational-only simulations found that AL 288-1 was unable to extend the hip like modern humans, forcing the specimen to employ a BHBK style of walking, thus contradicting 40+ years of previous research into the locomotory capabilities of AL 288-1. Therefore, we advocate that differences in the sphericity of the AL 288-1 femoral head with that of a modern human necessitates all six DOFs to be included in which AL 288-1 could osteologically extend the hip to facilitate a human-like gait.

A Guide to Inverse Kinematic Marker-Guided Rotoscoping Using IK Solvers.

X-ray Reconstruction of Moving Morphology (XROMM) permits researchers to see beneath the skin, usually to see musculoskeletal movements. These movements can be tracked and later used to provide information regarding the mechanics of movement. Here, we discuss "IK marker-guided rotoscoping"-a method that combines inverse kinematic solvers with that of traditional scientific rotoscoping methods to quickly and efficiently overlay 3D bone geometries with the X-ray shadows from XROMM data. We use a case study of three Nile crocodiles' (Crocodylus niloticus) forelimbs and hindlimbs to evaluate this method. Within these limbs, different marker configurations were used: some configurations had six markers, others had five markers, and all forelimb data only had three markers. To evaluate IK marker-guided rotoscoping, we systematically remove markers in the six-marker configuration and then test the magnitudes of deviation in translations and rotations of the rigged setup with fewer markers versus those of the six-marker configuration. We establish that IK marker-guided rotoscoping is a suitable method for "salvaging" data that may have too few markers.

La reconstruction par rayons X de la morphologie en mouvement (XROMM) permet aux chercheurs d'observer les structures internes d'un organisme en mouvement, notamment au niveau musculo-squelettique. Ces mouvements peuvent être suivis à l'aide de marqueurs, renseignant ainsi sur la mécanique des mouvements de l'organisme étudié. Nous présentons ici la méthode  'IK marker-guided rotoscoping' qui combine des solveurs de cinématique inverse (IK) et des méthodes de rotoscopie scientifique pour superposer efficacement et de manière fiable des os en 3D avec les données obtenues par XROMM. Afin d'estimer la précision de cette méthode, nous l'avons appliquée sur un échantillon composé de membres antérieurs et postérieurs de trois crocodiles du Nil (Crocodylus niloticus) en utilisant trois configurations différentes : certaines comprenaient six marqueurs, d'autres cinq, tandis que toutes les configurations sur le membre antérieur comprenaient trois marqueurs. Nous avons ensuite retiré des marqueurs dans toutes les configurations comprenant six marqueurs afin de vérifier si cette différence avait un impact sur les mouvements de translations et de rotations obtenus respectivement avec ces deux configurations. Nous démontrons ainsi qu'il est possible d'étudier des données incomplètes, et donc n'offrant que très peu de marqueurs, en utilisant la méthode 'IK marker-guided rotoscoping'.(by Romain Pintore).

Three-dimensional polygonal muscle modelling and line of action estimation in living and extinct taxa.

Biomechanical models and simulations of musculoskeletal function rely on accurate muscle parameters, such as muscle masses and lines of action, to estimate force production potential and moment arms. These parameters are often obtained through destructive techniques (i.e., dissection) in living taxa, frequently hindering the measurement of other relevant parameters from a single individual, thus making it necessary to combine multiple specimens and/or sources. Estimating these parameters in extinct taxa is even more challenging as soft tissues are rarely preserved in fossil taxa and the skeletal remains contain relatively little information about the size or exact path of a muscle. Here we describe a new protocol that facilitates the estimation of missing muscle parameters (i.e., muscle volume and path) for extant and extinct taxa. We created three-dimensional volumetric reconstructions for the hindlimb muscles of the extant Nile crocodile and extinct stem-archosaur Euparkeria, and the shoulder muscles of an extant gorilla to demonstrate the broad applicability of this methodology across living and extinct animal clades. Additionally, our method can be combined with surface geometry data digitally captured during dissection, thus facilitating downstream analyses. We evaluated the estimated muscle masses against physical measurements to test their accuracy in estimating missing parameters. Our estimated muscle masses generally compare favourably with segmented iodine-stained muscles and almost all fall within or close to the range of observed muscle masses, thus indicating that our estimates are reliable and the resulting lines of action calculated sufficiently accurately. This method has potential for diverse applications in evolutionary morphology and biomechanics.

One size fits all? Stature estimation from footprints and the effect of substrate and speed on footprint creation.

Estimation of stature from footprint lengths is a common prediction in forensic cases and in paleoanthropology upon the discovery of fossil footprints. Many studies, which have estimated stature from footprints, generally use a "one-size-fits-all" approach that usually involves applying a known ratio of foot length to total stature to do so, although this method has fallen out of practice in forensic cases in recent years but is still commonly used for fossil trace evidence. Yet, we know that substrate and speed can change the dimensions of a footprint, so why are these "one-size-fits-all" approaches still used today? We tested footprint production across different substrates at a walk, a fast walk, and a jog. We calculated how accurately footprint dimensions were impressed between these different conditions and identified sources of error in footprint lengths, and the percentage changes of how significantly a footprint can change in length between different conditions. We provide a table with different ratios that we encourage practitioners/field scientists to refer to and use when estimating stature from footprints, with respect to the substrate on which the footprint was created and the speed at which it was created. We actively encourage researchers to add the ratios by testing more substrates so that in the future stature can be more accurately estimated, thus aiding the paleoanthropological community, but also forensic investigations by statistically highlighting how different conditions can affect trace dimensions.

Musculoskeletal modelling of the Nile crocodile (Crocodylus niloticus) hindlimb: Effects of limb posture on leverage during terrestrial locomotion.

We developed a three-dimensional, computational biomechanical model of a juvenile Nile crocodile (Crocodylus niloticus) pelvis and hindlimb, composed of 47 pelvic limb muscles, to investigate muscle function. We tested whether crocodiles, which are known to use a variety of limb postures during movement, use limb orientations (joint angles) that optimise the moment arms (leverages) or moment-generating capacities of their muscles during different limb postures ranging from a high walk to a sprawling motion. We also describe the three-dimensional (3D) kinematics of the crocodylian hindlimb during terrestrial locomotion across an instrumented walkway and a treadmill captured via X-ray Reconstruction of Moving Morphology (biplanar fluoroscopy; 'XROMM'). We reconstructed the 3D positions and orientations of each of the hindlimb bones and used dissection data for muscle lines of action to reconstruct a focal, subject-specific 3D musculoskeletal model. Motion data for different styles of walking (a high, crouched, bended and two types of sprawling motion) were fed into the 3D model to identify whether any joints adopted near-optimal poses for leverage across each of the behaviours. We found that (1) the hip adductors and knee extensors had their largest leverages during sprawling postures and (2) more erect postures typically involved greater peak moment arms about the hip (flexion-extension), knee (flexion) and metatarsophalangeal (flexion) joints. The results did not fully support the hypothesis that optimal poses are present during different locomotory behaviours because the peak capacities were not always reached around mid-stance phase. Furthermore, we obtained few clear trends for isometric moment-generating capacities. Therefore, perhaps peak muscular leverage in Nile crocodiles is instead reached either in early/late stance or possibly during swing phase or other locomotory behaviours that were not studied here, such as non-terrestrial movement. Alternatively, our findings could reflect a trade-off between having to execute different postures, meaning that hindlimb muscle leverage is not optimised for any singular posture or behaviour. Our model, however, provides a comprehensive set of 3D estimates of muscle actions in extant crocodiles which can form a basis for investigating muscle function in extinct archosaurs.

The morphological affinity of the Early Pleistocene footprints from Happisburgh, England, with other footprints of Pliocene, Pleistocene, and Holocene age.

Fossil hominin footprints provide a direct source of evidence of locomotor behavior and allow inference of other biological data such as anthropometrics. Many recent comparative analyses of hominin footprints have used 3D analytical methods to assess their morphological affinities, comparing tracks from different locations and/or time periods. However, environmental conditions can sometimes preclude 3D digital capture, as was the case at Happisburgh (England) in 2013. Consequently, we use here a 2D geometric morphometric approach to investigate the evolutionary context of the Happisburgh tracks. The comparative sample of hominin tracks comes from eight localities that span a broad temporal range from the Pliocene to Late Holocene. The results show disparity in the shapes of tracks ascribed to hominins from the Pliocene (presumably Australopithecus afarensis), Pleistocene (presumably Homo erectus and Homo antecessor), and Holocene (Homo sapiens). Three distinct morphological differences are apparent between time samples: changes in adduction of the hallux, changes in the shape and position of the medial longitudinal arch impression, and apparent changes in foot proportions. Linear dimensions classified the potential H. antecessor tracks from Happisburgh as being most similar to the presumed H. erectus prints from Ileret. We demonstrate using 2D geometric morphometric methods and linear dimensions that the Happisburgh tracks are morphologically similar to other presumed Homo tracks and differ from the Laetoli footprints. The probable functional implications of these results fit well with previous comparative analyses of hominin tracks at other sites.