Identifying the neural correlates of anticipatory postural control: A novel fMRI paradigm.

Altered postural control in the trunk/hip musculature is a characteristic of multiple neurological and musculoskeletal conditions. Previously it was not possible to determine if altered cortical and subcortical sensorimotor brain activation underlies impairments in postural control. This study used a novel fMRI-compatible paradigm to identify the brain activation associated with postural control in the trunk and hip musculature. BOLD fMRI imaging was conducted as participants performed two versions of a lower limb task involving lifting the left leg to touch the foot to a target. For the supported leg raise (SLR) the leg is raised from the knee while the thigh remains supported. For the unsupported leg raise (ULR) the leg is raised from the hip, requiring postural muscle activation in the abdominal/hip extensor musculature. Significant brain activation during the SLR task occurred predominantly in the right primary and secondary sensorimotor cortical regions. Brain activation during the ULR task occurred bilaterally in the primary and secondary sensorimotor cortical regions, as well as cerebellum and putamen. In comparison with the SLR, the ULR was associated with significantly greater activation in the right premotor/SMA, left primary motor and cingulate cortices, primary somatosensory cortex, supramarginal gyrus/parietal operculum, superior parietal lobule, cerebellar vermis, and cerebellar hemispheres. Cortical and subcortical regions activated during the ULR, but not during the SLR, were consistent with the planning, and execution of a task involving multisegmental, bilateral postural control. Future studies using this paradigm will determine mechanisms underlying impaired postural control in patients with neurological and musculoskeletal dysfunction.

T1rho and T2 mapping of ankle cartilage of female and male ballet dancers.

BACKGROUND: Since ballet dancers begin their training before skeletal maturity, accurate and non-invasive identification of cartilage diseases is clinically important. Angle-dependent analysis of T1rho and T2 sequences can be useful for quantification of the composition of cartilage. PURPOSE: To investigate the angle-dependent T1rho and T2 profiles of ankle cartilage in non-dancers and dancers. MATERIAL AND METHODS: Ten female non-dancers, ten female dancers, and 9 male dancers were evaluated using T1rho and T2 mapping sequences. Manual segmentation of talar and tibial cartilage on these images was performed by two radiologists. Inter- and intra-rater reliabilities were calculated using intraclass correlation coefficients (ICCs) and Bland-Altman analysis. Mean thickness and volume of cartilage were estimated. Angle-dependent relaxation time profiles of talar and tibial cartilage were created. RESULTS: ICCs of the number of segmented pixels were poor to excellent. Bland-Altman plots indicated that differences were associated with segment sizes. Segmented cartilage on T1rho demonstrated larger thickness and volume than those on T2 in all populations. Male dancers showed larger cartilage thickness and volume than female dancers and non-dancers. Each cartilage demonstrated angular-dependent T1rho and T2 profiles. Minimal T1rho and T2 values were observed at approximately 180°-200°; higher values were seen at the angle closer to the magic angle. Minimal T2 value of talar cartilage of dancers was larger than that of non-dancers. CONCLUSION: In this small cohort study, regional and sex variations of ankle cartilage T1rho and T2 values in dancers and non-dancers were demonstrated using an angle-dependent approach.

Central Mechanisms Mediating Thrombospondin-4-induced Pain States.

Peripheral nerve injury induces increased expression of thrombospondin-4 (TSP4) in spinal cord and dorsal root ganglia that contributes to neuropathic pain states through unknown mechanisms. Here, we test the hypothesis that TSP4 activates its receptor, the voltage-gated calcium channel Cavα2δ1 subunit (Cavα2δ1), on sensory afferent terminals in dorsal spinal cord to promote excitatory synaptogenesis and central sensitization that contribute to neuropathic pain states. We show that there is a direct molecular interaction between TSP4 and Cavα2δ1 in the spinal cord in vivo and that TSP4/Cavα2δ1-dependent processes lead to increased behavioral sensitivities to stimuli. In dorsal spinal cord, TSP4/Cavα2δ1-dependent processes lead to increased frequency of miniature and amplitude of evoked excitatory post-synaptic currents in second-order neurons as well as increased VGlut2- and PSD95-positive puncta, indicative of increased excitatory synapses. Blockade of TSP4/Cavα2δ1-dependent processes with Cavα2δ1 ligand gabapentin or genetic Cavα2δ1 knockdown blocks TSP4 induced nociception and its pathological correlates. Conversely, TSP4 antibodies or genetic ablation blocks nociception and changes in synaptic transmission in mice overexpressing Cavα2δ1 Importantly, TSP4/Cavα2δ1-dependent processes also lead to similar behavioral and pathological changes in a neuropathic pain model of peripheral nerve injury. Thus, a TSP4/Cavα2δ1-dependent pathway activated by TSP4 or peripheral nerve injury promotes exaggerated presynaptic excitatory input and evoked sensory neuron hyperexcitability and excitatory synaptogenesis, which together lead to central sensitization and pain state development.

Characterization of ectopic colonies that form in widespread areas of the nervous system with neural stem cell transplants into the site of a severe spinal cord injury.

We reported previously the formation of ectopic colonies in widespread areas of the nervous system after transplantation of fetal neural stem cells (NSCs) into spinal cord transection sites. Here, we characterize the incidence, distribution, and cellular composition of the colonies. NSCs harvested from E14 spinal cords from rats that express GFP were treated with a growth factor cocktail and grafted into the site of a complete spinal cord transection. Two months after transplant, spinal cord and brain tissue were analyzed histologically. Ectopic colonies were found at long distances from the transplant in the central canal of the spinal cord, the surface of the brainstem and spinal cord, and in the fourth ventricle. Colonies were present in 50% of the rats, and most rats had multiple colonies. Axons extended from the colonies into the host CNS. Colonies were strongly positive for nestin, a marker for neural precursors, and contained NeuN-positive cells with processes resembling dendrites, GFAP-positive astrocytes, APC/CC1-positive oligodendrocytes, and Ki-67-positive cells, indicating ongoing proliferation. Stereological analyses revealed an estimated 21,818 cells in a colony in the fourth ventricle, of which 1005 (5%) were Ki-67 positive. Immunostaining for synaptic markers (synaptophysin and VGluT-1) revealed large numbers of synaptophysin-positive puncta within the colonies but fewer VGluT-1 puncta. Continuing expansion of NSC-derived cell masses in confined spaces in the spinal cord and brain could produce symptoms attributable to compression of nearby tissue. It remains to be determined whether other cell types with self-renewing potential can also form colonies.

Calcium channel α2δ1 proteins mediate trigeminal neuropathic pain states associated with aberrant excitatory synaptogenesis.

To investigate a potential mechanism underlying trigeminal nerve injury-induced orofacial hypersensitivity, we used a rat model of chronic constriction injury to the infraorbital nerve (CCI-ION) to study whether CCI-ION caused calcium channel α2δ1 (Cavα2δ1) protein dysregulation in trigeminal ganglia and associated spinal subnucleus caudalis and C1/C2 cervical dorsal spinal cord (Vc/C2). Furthermore, we studied whether this neuroplasticity contributed to spinal neuron sensitization and neuropathic pain states. CCI-ION caused orofacial hypersensitivity that correlated with Cavα2δ1 up-regulation in trigeminal ganglion neurons and Vc/C2. Blocking Cavα2δ1 with gabapentin, a ligand for the Cavα2δ1 proteins, or Cavα2δ1 antisense oligodeoxynucleotides led to a reversal of orofacial hypersensitivity, supporting an important role of Cavα2δ1 in orofacial pain processing. Importantly, increased Cavα2δ1 in Vc/C2 superficial dorsal horn was associated with increased excitatory synaptogenesis and increased frequency, but not the amplitude, of miniature excitatory postsynaptic currents in dorsal horn neurons that could be blocked by gabapentin. Thus, CCI-ION-induced Cavα2δ1 up-regulation may contribute to orofacial neuropathic pain states through abnormal excitatory synapse formation and enhanced presynaptic excitatory neurotransmitter release in Vc/C2.

Effect of overground training augmented by mental practice on gait velocity in chronic, incomplete spinal cord injury.

OBJECTIVE: To compare the efficacy of a regimen combining mental practice (MP) with overground training (OT) with the efficacy of a regimen consisting of OT only on gait velocity and lower extremity motor outcomes in individuals with chronic (>12mo postinjury), incomplete spinal cord injury (SCI). DESIGN: Randomized, controlled, single-blinded study. SETTING: Outpatient rehabilitation laboratories. PARTICIPANTS: Subjects with chronic, incomplete SCI (N=18). INTERVENTIONS: Subjects were randomly assigned to receive (1) OT only, occurring 3d/wk for 8 weeks; or (2) OT augmented by MP (MP + OT), during which randomly assigned subjects listened to an MP audio recording directly after OT sessions. MAIN OUTCOME MEASURES: Subjects were administered a test of gait velocity as well as the Tinetti Performance Oriented Mobility Assessment, Spinal Cord Injury Independence Measure, and Satisfaction With Life Scale on 2 occasions before intervention, 1 week after intervention, and 12 weeks after intervention. RESULTS: A significant increase in gait velocity was exhibited across subjects at both 1 week posttherapy (P=.005) and at 12 weeks posttherapy (P=.006). However, no differences were seen in intervention response at either 1 or 12 weeks postintervention among subjects in the MP + OT group versus the OT-only group. CONCLUSIONS: OT was associated with significant gains in gait velocity, and these gains were not augmented by further addition of MP.

Thrombospondin-4 contributes to spinal sensitization and neuropathic pain states.

Neuropathic pain is a common cause of pain after nerve injury, but its molecular basis is poorly understood. In a post-gene chip microarray effort to identify new target genes contributing to neuropathic pain development, we report here the characterization of a novel neuropathic pain contributor, thrombospondin-4 (TSP4), using a neuropathic pain model of spinal nerve ligation injury. TSP4 is mainly expressed in astrocytes and significantly upregulated in the injury side of dorsal spinal cord that correlates with the development of neuropathic pain states. TSP4 blockade by intrathecal antibodies, antisense oligodeoxynucleotides, or inactivation of the TSP4 gene reverses or prevents behavioral hypersensitivities. Intrathecal injection of TSP4 protein into naive rats is sufficient to enhance the frequency of EPSCs in spinal dorsal horn neurons, suggesting an increased excitatory presynaptic input, and to cause similar behavioral hypersensitivities. Together, these findings support that injury-induced spinal TSP4 may contribute to spinal presynaptic hypersensitivity and neuropathic pain states. Development of TSP4 antagonists has the therapeutic potential for target-specific neuropathic pain management.

Feasibility of home hand rehabilitation using musicglove after chronic spinal cord injury.

STUDY DESIGN: Randomized, controlled single-blind cross over study. This study was registered on ClinicalTrials.gov (NCT02473614). OBJECTIVES: Examine usership patterns and feasibility of MusicGlove for at home hand rehabilitation therapy following chronic spinal cord injury. SETTING: Homes of participants. METHODS: Ten participants with chronic spinal cord injury completed two baseline assessments of hand function. After a stable baseline was determined all participants were randomized into two groups: Experimental and Control. Each group was given a recommended therapy dosage. Following this participants switched interventions. RESULTS: On average participants had higher levels of compliance (6.1 ± 3.5 h.), and completed more grips (15,760 ± 9,590 grips) compared to participants in previous stroke studies using the same device. Participants modulated game parameters in a manner consistent with optimal challenge principles from motor learning theory. Participants in the experimental group increased their prehension ability (1 ± 1.4 MusicGlove, 0.2 ± 0.5 Control) and performance (1.4 ± 2.2 MusicGlove, 0.4 ± 0.55 Control) on the Graded and Redefined Assessment of Strength, Sensibility, and Prehension subtests. Increases in performance on the Box and Blocks Test also favored the experimental group compared to the conventional group at the end of therapy (4.2 ± 5.9, -1.0 ± 3.4 respectively). CONCLUSIONS: MusicGlove is a feasible option for hand therapy in the home-setting for individuals with chronic SCI. Participants completed nearly twice as many gripping movements compared to individuals from the sub-acute and chronic stroke populations, and a number far greater than the number of movements typically achieved during traditional rehabilitation.

Quantitative MRI Analysis of the Talocrural and Talonavicular Joints in Ballet Dancers.

The ankles of ballet dancers are routinely under heavy loading that may lead to osteoarthritic changes. It would be clinically useful to identify such pathology as early as possible in a dancer's career. Therefore, the purpose of this study was to compare quantitative measurements in magnetic resonance (MR) images of the talocrural and talonavicular joints in ballet dancers and healthy non-dancers for use in formulating prediction of chronic injury and degenerative joint disease in these locations. Quantitative measurements in MR images of the talocrural and talonavicular joints were compared in 10 female ballet dancers, 10 healthy female non-dancers, and nine male ballet dancers. Fat-suppressed density-weighted proton, T1rho, and T2 mapping images were acquired with a 3.0 T MR scanner. Medial and lateral subchondral bone distance between the tibia and talus (MSBD and LSBD), axial navicular-talus axis angle (ANT angle), sagittal talar neck angle against the posterior talocalcaneal joint (TN angle), and curvature of navicular surface at the talonavicular joint were measured on sagittal images. The medial subchondral bone distance was found to be significantly larger in female dancers than female non-dancers (4.05 mm vs. 2.75 mm, p < 0.05), whereas there were no significant differences in LSBD (2.63 mm vs. 2.63 mm, p = 0.87). Axial navicular talus angles in female dancers were significantly larger than those in female non-dancers (38.9° vs. 24.3°, p < 0.05). There was a tendency for the TN angle to be smaller and navicular curvature (NC) to be larger in female dancers compared to female non-dancers, though the differences were not significant (TN angle: 16.6° vs. 22.3°, p = 0.09, and NC: 0.186 vs. 0.165, p = 0.28). There were no significant differences in T1rho or T2 values of talonavicular joint cartilage. These results show that the bony anatomy of dancers' ankles may adapt to the stresses placed on them by ballet.

Regenerative growth of corticospinal tract axons via the ventral column after spinal cord injury in mice.

Studies that have assessed regeneration of corticospinal tract (CST) axons in mice after genetic modifications or other treatments have tacitly assumed that there is little if any regeneration of CST axons in normal mice in the absence of some intervention. Here, we document a previously unrecognized capability for regenerative growth of CST axons in normal mice that involves growth past the lesion via the ventral column. Mice received dorsal hemisection injuries at thoracic level 6-7, which completely transect descending CST axons in the dorsal and dorsolateral column. Corticospinal projections were traced by injecting biotinylated dextran amine (BDA) into the sensorimotor cortex of one hemisphere either at the time of the injury or 4 weeks after injury, and mice were killed at 20-23 or 46 d after injury. At 20-23 d after injury, BDA-labeled CST axons did not extend past the lesion except in one animal. By 46 d after injury, however, a novel population of BDA-labeled CST axons could be seen extending from the gray matter rostral to the injury into the ventral column, past the lesion, and then back into the gray matter caudal to the injury in which they formed elaborate terminal arbors. The number of axons with this highly unusual trajectory was small ( approximately 1% of the total number of labeled CST axons rostral to the injury). The BDA-labeled axons in the ventral column were on the same side as the main tract and thus are not spared ventral CST axons (which would be contralateral to the main tract). These results indicate that normal mice have a capacity for CST regeneration that has not been appreciated previously, which has important implications in studying the effect of genetic or pharmacological manipulations on CST regeneration in mice.

A Six Week Therapeutic Ballet Intervention Improved Gait and Inhibitory Control in Children With Cerebral Palsy-A Pilot Study.

Children with cerebral palsy (CP) have motor impairments that make it challenging for them to participate in standard physical activity (PA) interventions. There is a need to evaluate adapted PA interventions for this population. Dance can promote coordination, posture, muscle strength, motor learning, and executive functioning. This pilot study evaluated the feasibility and the effects of a new therapeutic ballet intervention specifically designed for children with CP. Methods: Eight children with CP (9-14 y/o; 75% female) participated in a 6-week therapeutic ballet intervention. Outcomes were measured in multiple domains, including body composition (DXA), muscle strength (hand-grip dynamometer), habitual physical activity, gait and selective motor control functions, and executive functioning. Follow-up assessments of habitual physical activity, gait, and executive functioning were completed 4 to 5 weeks post-intervention. Results: Five of the eight participants were overfat or obese based on DXA percentage of body fat. All participants were below the 50th percentile for their age and gender for bone density. Four participants showed a trend to improve hand-grip strength in one hand only, while one improved in both hands. There were significant improvements in gait across time points (pre, post, and follow-up), specifically in time of ambulation (X pre = 4.36, X post = 4.22, X follow-up = 3.72, d = 0.056, p = 0.02), and in step length (cm) on the right: X pre = 48.29, X post = 50.77, X follow-up = 52.11, d = 0.22, p = 0.027, and left stride: X pre = 96.29, X post = 102.20, X follow-up = 104.20, d = 0.30, p = 0.027, indicating gait changes in bilateral lower extremities. There was improvement in inhibitory control (d = 0.78; 95% Confidence Limit = ±0.71, p < 0.05) with large individual responses primarily among those above the mean at baseline. Conclusions: Therapeutic ballet may prove to be a useful intervention to promote physiological and cognitive functions in children with CP. Results demonstrated feasibility of the physical, physiological, and cognitive assessments and suggested improvements in participants' gait and inhibitory control with large individual responses. Modifications to personalize the intervention may be needed to optimize positive outcomes. Clinical Trial Registration: www.ClinicalTrials.gov, identifier: NCT03681171.

Robot-assisted hindlimb extension increases the probability of swing initiation during treadmill walking by spinal cord contused rats.

Training and evaluation of locomotion in animals with spinal cord injury will likely be improved with the development of techniques that increase stepping activity. We hypothesized that robot-assisted extension of the hindlimbs of spinal cord injured rats during stance would increase the probability that the swing phase of gait would be initiated. Thirty-three adult, Sprague-Dawley rats received a contusion injury to the mid-thoracic spinal cord. The animals' hindlimbs were pulled into extension using small robotic arms to pull at the ankle, as the rat stepped on either a reciprocating, robotic paw platform or a conventional treadmill belt. The animals demonstrated an increase in the probability of swing initiation with spontaneous recovery during the first 4 weeks following injury (p < 0.0001). The probability of swing initiation was found to be greater with the use of robot-assisted extension versus no extension force at the ankle (75+/-16.9% versus 38.9+/-16.6%, p < 0.001). Swing initiation occurred most frequently at a particular distance of hindlimb extension (50 mm caudal from the neutral position at stance), but was broadly tuned about this distance. These results indicate that a greater amount of stepping activity can be elicited by robot-assisted extension, thereby providing possible benefits to evaluation and training of gait following SCI.

Increased Brain Sensorimotor Network Activation after Incomplete Spinal Cord Injury.

After complete spinal cord injury (SCI), activation during attempted movement of paralyzed limbs is sharply reduced, but after incomplete SCI-the more common form of human injury-it is unknown how attempts to move voluntarily are accompanied by activation of brain motor and sensory networks. Here, we assessed brain activation during ankle movement in subjects with incomplete SCI, among whom voluntary motor function is partially preserved. Adults with incomplete SCI (n = 20) and healthy controls (n = 15) underwent functional magnetic resonance imaging that alternated rest with 0.3-Hz right ankle dorsiflexion. In both subject groups, ankle movement was associated with bilateral activation of primary and secondary sensory and motor areas, with significantly (p < 0.001) greater activation in subjects with SCI within right hemisphere areas, including primary sensorimotor cortex and pre-motor cortex. This result was further evaluated using linear regression analysis with respect to core clinical variables. Poorer locomotor function correlated with larger activation within several right hemisphere areas, including pre- and post-central gyri, possibly reflecting increased movement complexity and effort, whereas longer time post-SCI was associated with larger activation in left post-central gyrus and bilateral supplementary motor area, which may reflect behaviorally useful adaptations. The results indicate that brain adaptations after incomplete SCI differ sharply from complete SCI, are related to functional behavioral status, and evolve with increasing time post-SCI. The results suggest measures that might be useful for understanding and treating incomplete SCI in human subjects.

Robotic gait analysis of bipedal treadmill stepping by spinal contused rats: characterization of intrinsic recovery and comparison with BBB.

There is a critical need to develop objective, quantitative techniques to assess motor function after spinal cord injury. Here, we assess the ability of a recently developed robotic device (the "rat stepper") to characterize locomotor impairment following contusion injury in rats. In particular, we analyzed how the kinematic features of hindlimb movement during bipedal, weight-supported treadmill stepping change following contusion, and whether these changes correlate with the recovery of open field locomotion. Female, Sprague-Dawley rats (n=29, 8 weeks of age) received mid thoracic contusion injuries of differing severities (11 mild, nine moderate, nine severe, and four sham). In a first experiment, 16 of the animals were evaluated weekly for 12 weeks using the robotic stepping device. In a second experiment, 17 of the animals were evaluated every other day for 4 weeks. The contused animals recovered open field locomotion based on the Basso, Beattie, and Bresnahan Scale (BBB) analysis, with most of the recovery occurring by 4 weeks post-injury. Analysis of 14 robotic measures of stepping revealed that several measures improved significantly during the same 4 weeks: swing velocity, step height, step length, hindlimb coordination, and the ability to support body weight. These measures were also significantly correlated with the BBB score. The number of steps taken during testing was not directly related to intrinsic recovery or correlated to the BBB score. These results suggest that it is the quality of weight-supported steps, rather than the quantity, that best reflects locomotor recovery after contusion injury, and that the quality of these steps is determined by the integrity of extensor, flexor, and bilateral coordination pathways. Thus, by measuring only a few weight-supported steps with motion capture, a sensitive, valid measure of locomotor recovery following contusion injury can be obtained across a broad range of impairment levels.

Robotic Rehabilitator of the Rodent Upper Extremity: A System and Method for Assessing and Training Forelimb Force Production after Neurological Injury.

Rodent models of spinal cord injury are critical for the development of treatments for upper limb motor impairment in humans, but there are few methods for measuring forelimb strength of rodents, an important outcome measure. We developed a novel robotic device--the Robotic Rehabilitator of the Rodent Upper Extremity (RUE)--that requires rats to voluntarily reach for and pull a bar to retrieve a food reward; the resistance of the bar can be programmed. We used RUE to train forelimb strength of 16 rats three times per week for 23 weeks before and 38 weeks after a mild (100 kdyne) unilateral contusion at the cervical level 5 (C5). We measured maximum force produced when RUE movement was unexpectedly blocked. We compared this blocked pulling force (BPF) to weekly measures of forelimb strength obtained with a previous, well-established method: the grip strength meter (GSM). Before injury, BPF was 2.6 times higher (BPF, 444.6 ± 19.1 g; GSM, 168.4 ± 3.1 g) and 4.9 times more variable (p < 0.001) than pulling force measured with the GSM; the two measurement methods were uncorrelated (R(2) = 0.03; p = 0.84). After injury, there was a significant decrease in BPF of 134.35 g ± 14.71 g (p < 0.001). Together, our findings document BPF as a repeatable measure of forelimb force production, sensitive to a mild spinal cord injury, which comes closer to measuring maximum force than the GSM and thus may provide a useful measure for quantifying the effects of treatment in rodent models of SCI.

The Badges Program: A Self-Directed Learning Guide for Residents for Conducting Research and a Successful Peer-Reviewed Publication.

INTRODUCTION: The Badges Program is a self-directed supplement to a program's research curriculum. This step-by-step resource helps medical residents to understand the resources needed to conduct their own research project and fulfill the Accreditation Council for Graduate Medical Education requirements for scholarly activity. METHODS: The curriculum is completed over varying amounts of time depending on the intricacy of scholastic activities. Simple case reports may take as little as 1-2 weeks whereas months to years may be required if residents are completing more intricate and elaborate projects. Associated materials include a guide organized by topic followed by sequentially completed task assignments demonstrating the user has gained a basic understanding of the corresponding objective. Upon completing a task, the user obtains its educational badge, a virtual certificate of completion the program can then track with a simple checklist for progress. Several supplementary online articles review core concepts or provide examples and web-based tutorials for becoming proficient with using reference software. No faculty training is necessary. Residents go through the steps necessary to perform research so as to gain familiarity with the process. RESULTS: Qualitative feedback obtained by informally surveying residents who have completed the Badges Program has been very positive, with residents reporting that the guide was easy to follow and the knowledge gained will help tremendously with future projects. DISCUSSION: We hope that with feedback, this resource will be expanded and refined so every resident will be able to use the Badges Program regardless of previous skills, experience, and publications.

A re-assessment of treatment with a tyrosine kinase inhibitor (imatinib) on tissue sparing and functional recovery after spinal cord injury.

This study was undertaken as part of the NIH "Facilities of Research Excellence-Spinal Cord Injury" project to support independent replication of published studies. Here, we repeat key parts of a study reporting that rats treated with imatinib (Gleevec®, Novartis) after spinal cord contusion injury exhibited enhanced bladder function, greater recovery of motor function, and increased tissue sparing. Young adult female SCA Sprague-Dawley rats received moderate contusion injuries at T9-T10 using the MASCIS weight drop device. One group (n=16) received oral doses of imatinib 30min after injury and then daily doses for 5days. A control group (n=18) received vehicle. Motor function was assessed with the BBB locomotor rating scale and a contact plantar placement task. Bladder function was assessed by measuring the amount of urine retained in the bladder. Tissue preservation was assessed by immunostaining and stereological analysis. Rats that received imatinib had lower volumes of retained urine, suggesting improved bladder function, but there were no significant differences in motor function on any of the other tasks. Tissue preservation was assessed by immunostaining and stereological analysis. Quantitative analysis of spared tissue, cyst size, spared white matter, and inflammatory cell invasion revealed no significant differences between imatinib treated and control rats. Taken together our results confirm the findings that treatment with imatinib improves bladder function after SCI but fail to replicate findings of improved motor function, enhanced tissue sparing, and decreased inflammatory cell invasion.

A re-assessment of long distance growth and connectivity of neural stem cells after severe spinal cord injury.

As part of the NIH "Facilities of Research Excellence-Spinal Cord Injury" project to support independent replication, we repeated key parts of a study reporting robust engraftment of neural stem cells (NSCs) treated with growth factors after complete spinal cord transection in rats. Rats (n=20) received complete transections at thoracic level 3 (T3) and 2weeks later received NSC transplants in a fibrin matrix with a growth factor cocktail using 2 different transplantation methods (with and without removal of scar tissue). Control rats (n=9) received transections only. Hindlimb locomotor function was assessed with the BBB scale. Nine weeks post injury, reticulospinal tract axons were traced in 6 rats by injecting BDA into the reticular formation. Transplants grew to fill the lesion cavity in most rats although grafts made with scar tissue removal had large central cavities. Grafts blended extensively with host tissue obliterating the astroglial boundary at the cut ends, but in most cases there was a well-defined partition within the graft that separated rostral and caudal parts of the graft. In some cases, the partition contained non-neuronal scar tissue. There was extensive outgrowth of GFP labeled axons from the graft, but there was minimal ingrowth of host axons into the graft revealed by tract tracing and immunocytochemistry for 5HT. There were no statistically significant differences between transplant and control groups in the degree of locomotor recovery. Our results confirm the previous report that NSC transplants can fill lesion cavities and robustly extend axons, but reveal that most grafts do not create a continuous bridge of neural tissue between rostral and caudal segments.

A re-assessment of the effects of treatment with a non-steroidal anti-inflammatory (ibuprofen) on promoting axon regeneration via RhoA inhibition after spinal cord injury.

This study was undertaken as part of the NIH "Facilities of Research Excellence-Spinal Cord Injury" project to support independent replication of published studies. Here, we repeat key parts of a study reporting that rats treated with ibuprofen via subcutaneous minipump exhibited greater recovery of motor function and enhanced axonal growth after spinal cord injury. We carried out 3 separate experiments in which young adult female Sprague-Dawley rats received dorsal over-hemisections at T6-T7, and then were implanted with osmotic minipumps for subcutaneous delivery of ibuprofen or saline. Motor function was assessed with the BBB Locomotor Rating Scale, footprint analysis, and with a grid walk task. Combined group sizes for functional analyses were n=34 rats treated with ibuprofen and n=39 controls. Bladder function was assessed by measuring the amount of urine retained in the bladder twice per day. Four weeks post-injury, CST axons were traced by injecting BDA into the sensorimotor cortex; 5HT axons were assessed by immunostaining. Analysis of data from all rats revealed no significant differences between groups. Analysis of data excluding rats with lesions that were larger than intended indicated improved locomotor function in ibuprofen-treated rats at early post-lesion intervals in one of the individual experiments. Rats that received Ibuprofen did not demonstrate statistically significant improvements in bladder function. Quantitative analyses of CST and 5HT axon distribution also did not reveal differences between ibuprofen-treated and control rats. Taken together, our results only partially replicate the findings that treatment with ibuprofen improves motor function after SCI but fail to replicate findings regarding enhanced axon growth.