Hindlimb Movements Research Articles

Transplantation of peripheral nerve tissueoid based on a decellularized optic nerve scaffold to restore rat hindlimb sensory and movement functions

Transplantation of peripheral nerve tissueoid based on a decellularized optic nerve scaffold to restore rat hindlimb sensory and movement functions

Sonic Hedgehog reduces inflammatory response, decreases blood-spinal cord barrier permeability, and improves locomotor function recovery in an acute spinal cord injury rat model

BackgroundSonic Hedgehog (Shh), extensively researched for its role in early neurogenesis and brain development, has recently been recognized for its neuroprotective potential following neuronal injuries. This study examines the immediate impact of early administered Shh on the local inflammatory response post-acute spinal cord injury in rats.MethodsThirty-four female Wistar rats underwent either sham surgery (laminectomy; n = 10) or clip compression/contusion spinal cord injury (SCI) at the T9 level. This was followed by implantation of an osmotic pump and a subdural catheter for continuous intrathecal delivery of Shh (n = 12) or placebo (NaCl; n = 12). Locomotor function was assessed at 3- and 7-days post-injury (dpi) using the Basso, Beattie, and Bresnahan (BBB) score and the Gridwalk test. Animals were euthanized after 3 or 7 days for immunohistochemical analysis of the local inflammatory reaction and immune cell migration.ResultsShh-treated rats demonstrated significant hindlimb movement and coordination improvements at 7 days post-injury, compared to controls. This enhancement was accompanied by a significant reduction in both immune cell presence and blood plasma products within spinal cord lesions, suggesting Shh’s dual role in modulating immune cell migration and maintaining the integrity of the blood-spinal cord barrier. Separately, these Shh-treated rats also showed an increase in M(IL-4) polarization of macrophages, further underlining the potential therapeutic impact of Shh in post-injury recovery. Notably, these effects were not evident at three days post-injury.ConclusionShh application at 7 days post-injury showed immunomodulatory effects, possibly via enhanced blood-spinal cord barrier integrity, reduced immune cell migration, and increased anti-inflammatory immune cell differentiation. These mechanisms collectively contribute to enhanced locomotor recovery.

Relationship between COVID-19 and orofacial clefts.

MicroRNAs (miRNAs) are essential modulators of gene expression and are associated with various pathological processes, including spinal cord injury (SCI). This investigation aimed to elucidate miR-10a activity in SCI and its potential interaction with sirtuin 1 (SIRT1). The SCI rat model was established to assess hind limb movement, measure levels of miR-10a, SIRT1, neuronal survival, and inflammatory factors. An in-vitro SCI cell model was also developed to evaluate cell viability and inflammatory factor levels. The interaction between miR10a and SIRT1 was verified. Upregulated miR-10a and downregulated SIRT1 expression were found in the tissues of SCI rats. miR-10a knockdown in SCI rats enhanced the recovery of motor function, increased neuronal survival, and reduced the levels of inflammatory cytokines. Luciferase reporter assays confirmed that miR-10a targeted SIRT1 directly. In PC12 cells, downregulation of miR-10a increased SIRT1 expression, enhanced cell viability, and reduced inflammatory factor levels after LPS stimulation. Conversely, SIRT1 knockdown inhibited the protective effects of downregulated miR-10a on cell viability and inflammatory responses. The results suggest that miR-10a downregulation protects against SCI by upregulating SIRT1 expression, improving functional recovery, and reducing inflammation. Targeting the miR-10a/SIRT1 axis is a promising strategy for SCI treatment.

The LINGO-1-deficient neural stem cell-derived neural tissueoid showed enhanced retention and neuronal relay in the transected spinal cord

Neural stem cell (NSC)-derived neural tissueoids are a promising option to model spinal cord development and repair; however, the survival and integration ability with the host in vivo need to be improved. We aimed to determine if genetically engineered neural tissueoids in a NT-3 delivery scaffold exhibited increased retention and relay properties and to explore the underlying mechanisms. Gene expression screening of tissue and single-nucleus RNA sequencing revealed that leucine rich repeat and immunoglobin-like domain-containing protein 1 (LINGO-1) was significantly increased in neurons following spinal cord injury. The LINGO-1 protein accelerated cell death by increasing cleaved caspase-3 and interacting with TrkC to decrease NT-3-mediated phosphorylation of the PI3K/AKT/CREB axis, resulting in synaptic degradation. Subsequently, we seeded LINGO-1-deficient NSCs on a NT-3 delivery scaffold to construct an apoptosis resistant neural tissueoids, which was transplanted into a T9 complete spinal cord injury (SCI) rat model. The LINGO-1-deficient neural tissueoid was retained in the graft area, dramatically facilitated ascending and descending nerve fiber regeneration and connection, restored relay from brain to the caudal spinal cord, and improved the hindlimb movement function. This study demonstrates that transplantation of a LINGO-1-deficient tissueoid is an efficient pro-survival and neurogenesis strategy for the treatment of SCI.

Forelimb movements contribute to hindlimb cutaneous reflexes during locomotion in cats.

During quadrupedal locomotion, interactions between spinal and supraspinal circuits and somatosensory feedback coordinate forelimb and hindlimb movements. How this is achieved is not clear. To determine whether forelimb movements modulate hindlimb cutaneous reflexes involved in responding to an external perturbation, we stimulated the superficial peroneal nerve in six intact cats during quadrupedal locomotion and during hindlimb-only locomotion (with forelimbs standing on stationary platform) and in two cats with a low spinal transection (T12-T13) during hindlimb-only locomotion. We compared cutaneous reflexes evoked in six ipsilateral and four contralateral hindlimb muscles. Results showed similar occurrence and phase-dependent modulation of short-latency inhibitory and excitatory responses during quadrupedal and hindlimb-only locomotion in intact cats. However, the depth of modulation was reduced in the ipsilateral semitendinosus during hindlimb-only locomotion. Additionally, longer-latency responses occurred less frequently in extensor muscles bilaterally during hindlimb-only locomotion, whereas short-latency inhibitory and longer-latency excitatory responses occurred more frequently in the ipsilateral and contralateral sartorius anterior, respectively. After spinal transection, short-latency inhibitory and excitatory responses were similar to both intact conditions, whereas mid- or longer-latency excitatory responses were reduced or abolished. Our results in intact cats and the comparison with spinal-transected cats suggest that the absence of forelimb movements suppresses inputs from supraspinal structures and/or cervical cord that normally contribute to longer-latency reflex responses in hindlimb extensor muscles.NEW & NOTEWORTHY During quadrupedal locomotion, the coordination of forelimb and hindlimb movements involves central circuits and somatosensory feedback. To demonstrate how forelimb movement affects hindlimb cutaneous reflexes during locomotion, we stimulated the superficial peroneal nerve in intact cats during quadrupedal and hindlimb-only locomotion as well as in spinal-transected cats during hindlimb-only locomotion. We show that forelimb movement influences the modulation of hindlimb cutaneous reflexes, particularly the occurrence of long-latency reflex responses.

Removal of the infrapatellar fat pad and associated synovium benefits female guinea pigs in the Dunkin Hartley model of idiopathic osteoarthritis.

Several tissues contribute to the onset and advancement of knee osteoarthritis (OA). One tissue type that is worthy of closer evaluation, particularly in the context of sex, is the infrapatellar fat pad (IFP). We previously demonstrated that removal of the IFP had short-term beneficial effects for a cohort of male Dunkin-Hartley guinea pigs. The present project was designed to elucidate the influence of IFP removal in females of this OA-prone strain. It was hypothesized that resection of the IFP would reduce the development of OA in knees of a rodent model predisposed to the disease. Female guinea pigs (n=16) were acquired at an age of 2.5 months. Surgical removal of the IFP and associated synovium complex (IFP/SC) was executed at 3 months of age. One knee had the IFP/SC resected; a comparable sham surgery was performed on the contralateral knee. All animals were subjected to voluntary enclosure monitoring and dynamic weight-bearing, as well as compulsory treadmill-based gait analysis monthly; baseline data was collected prior to surgery. Guinea pigs were euthanized at 7 months. Knees from eight animals were evaluated via histology, mRNA expression, and immunohistochemistry (IHC); knees from the remaining eight animals were allocated to microcomputed tomography (microCT), biomechanical analyses (whole joint testing and indentation relaxation testing), and atomic absorption spectroscopy (AAS). Fibrous connective tissue (FCT) replaced the IFP/SC. Mobility/gait data indicated that unilateral IFP/SC removal did not affect bilateral hindlimb movement. MicroCT demonstrated that osteophytes were not a significant feature of OA in this sex; however, trabecular thickness (TbTh) in medial femorae decreased in knees containing the FCT. Histopathology scores were predominantly influenced by changes in the lateral tibia, which demonstrated that histologic signs of OA were increased in knees containing the native IFP/SC versus those with the FCT. Similarly, indentation testing demonstrated higher instantaneous and equilibrium moduli in the lateral tibial articular cartilage of control knees with native IFPs. AAS of multiple tissue types associated with the knee revealed that zinc was the major trace element influenced by removal of the IFP/SC. Our data suggest that the IFP/SC is a significant component driving knee OA in female guinea pigs and that resection of this tissue prior to disease has short-term benefits. Specifically, the formation of the FCT in place of the native tissue resulted in decreased cartilage-related OA changes, as demonstrated by reduced Osteoarthritis Research Society International (OARSI) histology scores, as well as changes in transcript, protein, and cartilage indentation analyses. Importantly, this model provides evidence that sex needs to be considered when investigating responses and associated mechanisms seen with this intervention.

Region-specific activation in the accumbens nucleus by itch with modified scratch efficacy in mice – a model-free multivariate analysis

Itch is a protective/defensive function with divalent motivational drives. Itch itself elicits an unpleasant experience, which triggers the urge to scratch, relieving the itchiness. Still, it can also result in dissatisfaction when the scratch is too intense and painful or unsatisfactory due to insufficient scratch effect. Therefore, it is likely that the balance between the unpleasantness/pleasure and satisfaction/unsatisfaction associated with itch sensation and scratching behavior is determined by complex brain mechanisms. The physiological/pathological mechanisms underlying this balance remain largely elusive. To address this issue, we targeted the "reward center" of the brain, the nucleus accumbens (NAc), in which itch-responsive neurons have been found in rodents. We examined how neurons in the NAc are activated or suppressed during histamine-induced scratching behaviors in mice. The mice received an intradermal injection of histamine or saline at the neck, and the scratching number was analyzed by recording the movement of the bilateral hind limbs for about 45 min after injection. To experimentally manipulate the scratch efficacy in these histamine models, we compared histamine's behavioral and neuronal effects between mice with intact and clipped nails on the hind paws. As expected, the clipping of the hind limb nail increased the number of scratches after the histamine injection. In the brains of mice exhibiting scratching behaviors, we analyzed the expression of the c-fos gene (Fos) as a readout of an immediate activation of neurons during itch/scratch and dopamine receptors (Drd1 and Drd2) using multiplex single-molecule fluorescence in situ hybridization (RNAscope) in the NAc and surrounding structures. We performed a model-free analysis of gene expression in geometrically divided NAc subregions without assuming the conventional core–shell divisions. The results indicated that even within the NAc, multiple subregions responded differentially to various itch/scratch conditions. We also found different clusters with neurons showing similar or opposite changes in Fos expression and the correlation between scratch number and Fos expression in different itch/scratch conditions. These regional differences and clusters would provide a basis for the complex role of the NAc and surrounding structures in encoding the outcomes of scratching behavior and itchy sensations.

Sustained delivery of NT-3 and curcumin augments microenvironment modulation effects of decellularized spinal cord matrix hydrogel for spinal cord injury repair.

Decellularized extracellular matrix hydrogel, especially that derived from spinal cord (DSCM hydrogel), has been actively considered as a functional biomaterial for remodeling the extracellular matrix of the native tissue, due to its unique characteristics in constructing pro-regenerative microenvironment for neural stem cells (NSCs). Furthermore, DSCM hydrogel can provide multiple binding domains to growth factors and drugs. Therefore, both exogenous neurotrophic factors and anti-inflammatory drugs are highly desired to be incorporated into DSCM hydrogel, which may synergistically modulate the complex microenvironment at the lesion site after spinal cord injury (SCI). Herein, neurotrophin-3 (NT-3) and curcumin (Cur) were integrated into DSCM hydrogel for SCI therapy. Due to different affinities to the DSCM hydrogel, NT-3 underwent a controlled release manner, while curcumin released explosively within the first 24 h, followed by rather sustained but slower release. The integration of both NT-3 and curcumin significantly enhanced NSCs proliferation and their neuronal differentiation. Meanwhile, the release of curcumin promoted macrophages polarization into anti-inflammatory subtypes, which further facilitated NSCs differentiation into neurons. The in situ injected DSCM + NT3 + Cur hydrogel exerted superior capability in alleviating the inflammatory responses in rat contused spinal cord. Compared to DSCM hydrogel alone, DSCM + NT3 + Cur hydrogel more significantly promoted the recruitment of NSCs and their neuronal differentiation at the lesion site. These outcomes favored functional recovery, as evidenced by the improved hind limb movement. Overall, the bioactive DSCM hydrogel can serve as a multifunctional carrier for cooperatively release of growth factors and drugs, which significantly benefits microenvironment regulation and nerve regeneration after SCI.

Inner and outer penetrating spinal cord injuries lead to distinct overground walking in mice

Spinal cord injury (SCI) is a devastating mechanical trauma. Although locomotion of model animals that mimic contusion SCI was actively examined, locomotion after penetrating SCI caused by sharp objects was not extensively studied. Severity of walking difficulty after partial transection of the spinal cord including penetrating SCI likely depends on the regions affected. Therefore, we compared beam walking and overground walking between mice after penetrating SCI at inner spinal cord region and mice with the injury at the outer region. Mice with the both penetrating SCIs did not display changes in beam walking. When appearance and movements of hindlimbs during overground walking was rated using Basso Mouse Scale for locomotion (BMS), however, mice with inner penetrating SCI showed low score shortly after the SCI. However, the score became high at later time points, as seen in contusion SCI mice. By contrast, BMS score did not decrease shortly after the outer penetrating SCI. However, the score became low 3 weeks after the SCI. As quantitative values during overground walking, movement duration in an open field were shorter at 1 day after the two penetrating SCIs. However, slower moving speed and fewer number of movement at 1 day were specific to mice with inner and outer penetrating SCIs, respectively. Moreover, BMS score was correlated with walking distance in open field only in mice with inner penetrating SCI. Thus, inner and outer penetrating SCI cause difficulty in overground walking with different severity and progress.

How otariids walk: a comparative axial analysis of the hind limb movements of California and South American sea lions

AbstractSeals exist at the intersection of land and water. This is especially true of eared seals (otariids), which are both highly adapted to forelimb swimming and retain the ability to walk on land. The precise nature and evolutionary history of their walking ability remains obscure, however. Here, we use axial analysis to compare the hind limb movements of California and South American sea lions with those of terrestrial plantigrades mammals. We show that, despite notable differences in foot orientation and joint position, all of our study species employ similar movements around the same centers of rotation. Our results suggest that otariid hind limbs and, in particular, the hallux play a substantial role in supporting and propelling the body during walking.

A bi-cortical neuroprosthesis to modulate locomotion after incomplete spinal cord injury.

Neuroprosthetic strategies seek to immediately alleviate deficits and reinstate voluntary control of movement. To facilitate recovery, it is crucial to gain a comprehensive understanding of the mechanisms involved in the return of intentional movement. Nevertheless, the precise relationship between the resurgence of cortical commands and the recovery of locomotion remains somewhat elusive. In the study conducted by Duguay, Bonizzato, Delivet-Mongrain, Fortier-Lebel and Martinez, we introduced a neuroprosthesis designed to deliver precise bi-cortical stimulation in a clinically relevant contusive spinal cord injury model. We conducted experiments in both healthy and spinal cord injured cats, where we fine-tuned the timing, duration, amplitude, and site of stimulation to modulate hindlimb locomotor output. In healthy cats, we observed a wide range of motor programs. However, after spinal cord injury, the induced hindlimb movements became highly stereotyped but were effective in modulating gait and reducing bilateral foot dragging. These results suggest that the neural basis for motor recovery traded off selectivity for effectiveness. Through a series of longitudinal assessments, we found that the restoration of locomotion following spinal cord injury was closely linked to the recovery of the descending neural drive. This underscores the importance of directing rehabilitation interventions toward the cortical target. The study results are discussed in terms of their impact and limitations.

Online repositories of photographs and videos provide insights into the evolution of skilled hindlimb movements in birds

The ability to manipulate objects with limbs has evolved repeatedly among land tetrapods. Several selective forces have been proposed to explain the emergence of forelimb manipulation, however, work has been largely restricted to mammals, which prevents the testing of evolutionary hypotheses in a comprehensive evolutionary framework. In birds, forelimbs have gained the exclusive function of flight, with grasping transferred predominantly to the beak. In some birds, the feet are also used in manipulative tasks and appear to share some features with manual grasping and prehension in mammals, but this has not been systematically investigated. Here we use large online repositories of photographs and videos to quantify foot manipulative skills across a large sample of bird species (>1000 species). Our results show that a complex interaction between niche, diet and phylogeny drive the evolution of manipulative skills with the feet in birds. Furthermore, we provide strong support for the proposition that an arboreal niche is a key element in the evolution of manipulation in land vertebrates. Our systematic comparison of foot use in birds provides a solid base for understanding morphological and neural adaptations for foot use in birds, and for studying the convergent evolution of manipulative skills in birds and mammals.

PH/Temperature Responsive Curcumin-Loaded Micelle Nanoparticles Promote Functional Repair after Spinal Cord Injury in Rats via Modulation of Inflammation.

The formation of an inhibitory inflammatory microenvironment after spinal cord injury (SCI) remains a great challenge for nerve regeneration. The poor local microenvironment exacerbates nerve cell death; therefore, the reconstruction of a favorable microenvironment through small-molecule drugs is a promising strategy for promoting nerve regeneration. In the present study, we synthesized curcumin-loaded micelle nanoparticles (Cur-NPs) to increase curcumin bioavailability and analyzed the physical and chemical properties of Cur-NPs by characterization experiments. We established an in vivo SCI model in rats and examined the ability of hind limb motor recovery using Basso-Beattie-Bresnahan scoring and hind limb trajectory assays. We also analyzed neural regeneration after SCI using immunofluorescence staining. The nanoparticles achieved the intelligent responsive release of curcumin while improving curcumin bioavailability. Most importantly, the released curcumin attenuated local inflammation by modulating the polarization of macrophages from an M1 pro-inflammatory phenotype to an M2 anti-inflammatory phenotype. M2-type macrophages can promote cell differentiation, proliferation, matrix secretion, and reorganization by secreting or expressing pro-repair cytokines to reduce the inflammatory response. The enhanced inflammatory microenvironment supported neuronal regeneration, nerve remyelination, and reduced scar formation. These effects facilitated functional repair in rats, mainly in the form of improved hindlimb movements. Here, we synthesized pH/temperature dual-sensitive Cur-NPs. While improving the bioavailability of the drug, they were also able to achieve a smart responsive release in the inflammatory microenvironment that develops after SCI. The Cur-NPs promoted the regeneration and functional recovery of nerves after SCI through anti-inflammatory effects, providing a promising strategy for the repair of SCIs.

Kinematic characteristics of canine hindlimb movement during sit-to-stand and stand-to-sit motions

Sit-to-stand and stand-to-sit motions are basic motions for daily animal life, and these motions are used as therapeutic exercises for dogs with functional impairments. The sit-to-stand motion is divided into several phases for kinesiological assessment in human rehabilitation and physical therapy. However, these motions in dogs have not been characterized in detail. We examined canine hindlimb kinematic characteristics during sit-to-stand/stand-to-sit motions and compared the characteristics with those during walking. In addition, we tried to classify phases of the movements based on kinematic characteristics of the transition of the range of motion of the hindlimb. We used a three-dimensional motion analysis system to evaluate the motions of eight clinically healthy beagles. During the sit-to-stand motion, the total range of motion (ROM) in the hip joint flexion/extension was half of that of during walking, but the total ROM of the hindlimb external/internal rotation relative to the pelvis and flexion/extension of the stifle and the tarsal joints were significantly larger than those of walking, suggesting that sit-to-stand exercise causes movements of hindlimb joints without marked changes in hip joint flexion/extension movement. Both sit-to-stand and stand-to-sit motions could not be divided into multiple phases only by the transition of the range of motion of the hindlimb.

Injectable Ventral Spinal Stimulator Evokes Programmable and Biomimetic Hindlimb Motion.

Spinal cord neuromodulation can restore partial to complete loss of motor functions associated with neuromotor disease and trauma. Current technologies have made substantial progress but have limitations as dorsal epidural or intraspinal devices that are either remote to ventral motor neurons or subject to surgical intervention in the spinal tissue. Here, we describe a flexible and stretchable spinal stimulator design with nanoscale thickness that can be implanted by minimally invasive injection through a polymeric catheter to target the ventral spinal space of mice. Ventrolaterally implanted devices exhibited substantially lower stimulation threshold currents and more precise recruitment of motor pools than did comparable dorsal epidural implants. Functionally relevant and novel hindlimb movements were achieved via specific stimulation patterns of the electrodes. This approach holds translational potential for improving controllable limb function following spinal cord injury or neuromotor disease.

Cerebellar axonopathy in Shivers horses identified by spatial transcriptomic and proteomic analyses.

Shivers in horses is characterized by abnormal hindlimb movement when walking backward and is proposed to be caused by a Purkinje cell (PC) axonopathy based on histopathology. Define region-specific differences in gene expression within the lateral cerebellar hemisphere and compare cerebellar protein expression between Shivers horses and controls. Case-control study of 5 Shivers and 4 control geldings ≥16.2 hands in height. Using spatial transcriptomics, gene expression was compared between Shivers and control horses in PC soma and lateral cerebellar hemisphere white matter, consisting primarily of axons. Tandem-mass-tag (TMT-11) proteomic analysis was performed on lateral cerebellar hemisphere homogenates. Differences in gene expression between Shivers and control horses were evident in principal component analysis of axon-containing white matter but not PC soma. In white matter, there were 455/1846 differentially expressed genes (DEG; 350 ↓DEG, 105 ↑DEG) between Shivers and controls, with significant gene set enrichment of the Toll-Like Receptor 4 (TLR4) cascade, highlighting neuroinflammation. There were 50/936 differentially expressed proteins (DEP). The 27 ↓DEP highlighted loss of axonal proteins including intermediate filaments (5), myelin (3), cytoskeleton (2), neurite outgrowth (2), and Na/K ATPase (1). The 23 ↑DEP were involved in the extracellular matrix (7), cytoskeleton (7), redox balance (2), neurite outgrowth (1), signal transduction (1), and others. Our findings support axonal degeneration as a characteristic feature of Shivers. Combined with histopathology, these findings are consistent with the known distinctive response of PC to injury where axonal changes occur without a substantial impact on PC soma.

Micromagnetic stimulation (µMS) dose-response of the rat sciatic nerve

Objective. The objective of this study was to investigate the effects of micromagnetic stimuli strength and frequency from the Magnetic Pen (MagPen) on the rat right sciatic nerve. The nerve’s response was measured by recording muscle activity and movement of the right hind limb. Approach. The MagPen was custom-built to be stably held over the sciatic nerve. Rat leg muscle twitches were captured on video, and movements were extracted using image processing algorithms. EMG recordings were also used to measure muscle activity. Main results. The MagPen prototype, when driven by an alternating current, generates a time-varying magnetic field, which, according to Faraday’s law of electromagnetic induction, induces an electric field for neuromodulation. The orientation-dependent spatial contour maps of the induced electric field from the MagPen prototype have been numerically simulated. Furthermore, in this in vivo work on µMS, a dose-response relationship has been reported by experimentally studying how varying the amplitude (Range: 25 mV p-p through 6 V p-p) and frequency (range: 100 Hz through 5 kHz) of the MagPen stimuli alters hind limb movement. The primary highlight of this dose-response relationship (repeated over n rats, where n = 7) is that for a µMS stimuli of higher frequency, significantly smaller amplitudes can trigger hind limb muscle twitch. This frequency-dependent activation can be justified by Faraday’s Law, which states that the magnitude of the induced electric field is directly proportional to the frequency. Significance. This work reports that µMS can successfully activate the sciatic nerve in a dose-dependent manner. The impact of this dose-response curve addresses the controversy in this research community about whether the stimulation from these μcoils arise from a thermal effect or micromagnetic stimulation. MagPen probes do not have a direct electrochemical interface with tissue and therefore do not experience electrode degradation, biofouling, and irreversible redox reactions like traditional direct contact electrodes. Magnetic fields from the μcoils create more precise activation than electrodes because they apply more focused and localized stimulation. Finally, unique features of µMS, such as the orientation dependence, directionality, and spatial specificity, have been discussed.

Reconstruction of soft noncontractile tissue in the derived therizinosaur Nothronychus: The interplay of soft tissue and stress on hindlimb ossification and posture.

Nothronychus graffami was a large therizinosaur from the Upper Cretaceous of North America. Much of the skeleton is well-preserved and relatively undistorted. The synovial capsule, extracapsular, and intracapsular tendons are reconstructed in N. graffami using existing scars and comparison with the hips of extant theropods and models of extinct theropods. The iliofemoral, pubofemoral, and ischiofemoral ligaments are all modeled. Soft tissue, especially the ischiofemoral and pubofemoral ligaments reduced possible protraction/retraction of the femur at the hip while stabilizing the joint. Therefore, most hindlimb movements took place at the knee. Weight-bearing function in Nothronychus was transferred from the supra-acetabular crest to the pubic peduncle and associated labrum at rest. The femur possessed an intermediately angled neck, convergent with titanosaurs. Therefore, some lateral abduction is proposed, requiring a well-developed meniscus at the knee. Such a posture would result in considerable mediolateral stress along the femoral shaft, resulting in increased transverse ossification. The femur is considered somewhat abducted from the midline, especially when maximally pronated, as the preacetabular ala is more extensive than the post-acetabular ala. This trait would probably result in a laterally divergent femur, as in many birds, resulting in a broad-gauge trackway with wider separation between left and right footprints than observed in many theropod trackways. Limb ratios and ossification patterns suggest a slow, waddling gait. A standard digitiform pose is reconstructed as is common for theropods, but a plantigrade stance cannot be firmly rejected.

189 Novel Wireless Battery-Free Fully Passive Flexible Brain/Spine Implant for Improved Recovery of Function Through Epidural Stimulation

INTRODUCTION: Acquired motor impairment from diseases such as stroke, trauma, epilepsy, and cancer remains an important cause of disability. Current treatments provide insufficient brain/spine recovery, with most patients failing to recover adequate motor function with rehabilitation alone. Brain stimulation can lead to enhanced synaptic density and synaptic response after long-term stimulation and enhanced motor function recovery. However, non-invasive options remain inadequate and invasive approaches are associated with unacceptable risks. METHODS: A flexible and transparent silicone elastomer substrate was used through microfabrication processes, with an external antenna near the implant to control the stimulation. In vitro validation was accomplished by wireless stimulation of human-induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs). In vivo validation was performed by motor cortex stimulation on an anesthetized rat. The corresponding hindlimb motion was captured by video. To separate implant-evoked movement from spontaneous movement, we compared the peak hindlimb velocity during 0.5 sec post-stimulation periods vs. the 0.5 sec pre-stimulation periods, across a continuous series of N = 43 stimulation pulses. RESULTS: Cultured hiPSC-CMs cells demonstrated well-controlled beating behavior under 0.5, 1, and 2Hz wireless stimulation. Statistical analysis further confirmed the change in beating rate corresponded with stimulation frequency (p < 0.001). The recorded hindlimb movement under wireless motor cortex stimulation demonstrated synchronization with the delivered voltage pulse trains. Average peak hindlimb velocity during pre-stimulation periods was 0.8 ± 0.4 mm/sec (± S.D.); during post-stimulation periods, 47 ± 13 mm/sec. A two-sample t-test confirmed the applied wireless power directly evoked movement, (p < 0.001). CONCLUSIONS: We have developed a wireless, battery-less implant with demonstrated efficacy in vitro and in vivo. Efforts at long-term safety evaluation are ongoing in a smaller and larger animal model as a bridge to phase 0/1 clinical trials.

Muscle architecture of the hindlimb of Tyto furcata (Aves Strigiformes): Highlights in owl morphology.

The American barn owl is a nocturnal bird of prey in which hind limb movements are a key factor in obtaining food; however, the architectural properties of its hind limb muscles have not been studied. This study sought to identify functional trends in the Tyto furcata hindlimb muscles by studying muscular architecture. The architectural parameters of the selected hip, knee, ankle, and digit muscles were studied in three specimens of the Tyto furcata and joint muscular proportions with an additional dataset were calculated. Previously published information on Asio otus was used for comparison. The flexor muscles of the digits had the highest muscle mass. Regarding architectural parameters, the main flexor of the digits (flexor digitorum longus) and the muscles that extend the knee and ankle joints (femorotibialis and gastrocnemius) showed a high physiological cross-sectional area (PCSA) and short fibers, allowing powerful digit flexion and knee and ankle extension. These mentioned features are in accordance with hunting behavior, in which prey capture is not only closely linked to the flexion of the digits but also to the movements of the ankle. During hunting, the distal hind limb is flexed and then fully extended at the moment of contact with the prey, whereas the digits are close to grasping the prey. The hip muscles showed a predominance of extensors over flexors, which were more massive, with parallel fibers and without tendons or short fibers. These features lead to a higher capacity to generate velocity to the detriment of forces, as indicated by the high values of the architectural index, their relatively low PCSA, and short or intermediate fiber length, which enhance the control of the joint positions and muscle length. Compared to Asio otus, Tyto furcata showed longer fibers; however, the relationship between fiber length and PCSA was similar for both species.