Quantification of prefrontal cortical neuronal ensembles following conditioned fear learning in a Fos-LacZ transgenic rat with photoacoustic imaging in Vivo.

Understanding the neurobiology of complex behaviors requires measurement of activity in the discrete population of active neurons, neuronal ensembles, which control the behavior. Conventional neuroimaging techniques ineffectively measure neuronal ensemble activity in the brain in vivo because they assess the average regional neuronal activity instead of the specific activity of the neuronal ensemble that mediates the behavior. Our functional molecular photoacoustic tomography (FM-PAT) system allows direct imaging of Fos-dependent neuronal ensemble activation in Fos-LacZ transgenic rats in vivo. We tested four experimental conditions and found increased FM-PAT signal in prefrontal cortical areas in rats undergoing conditioned fear or novel context exposure. A parallel immunofluorescence ex vivo study of Fos expression found similar findings. These findings demonstrate the ability of FM-PAT to measure Fos-expressing neuronal ensembles directly in vivo and support a mechanistic role for the prefrontal cortex in higher-order processing of response to specific stimuli or environmental cues.

Spatial and temporal dynamics of HDACs class IIa following mild traumatic brain injury in adult rats.

The fundamental role of epigenetic regulatory mechanisms involved in neuroplasticity and adaptive responses to traumatic brain injury (TBI) is gaining increased recognition. TBI-induced neurodegeneration is associated with several changes in the expression-activity of various epigenetic regulatory enzymes, including histone deacetylases (HDACs). In this study, PET/CT with 6-([18F]trifluoroacetamido)-1- hexanoicanilide ([18F]TFAHA) to image spatial and temporal dynamics of HDACs class IIa expression-activity in brains of adult rats subjected to a weight drop model of diffuse, non-penetrating, mild traumatic brain injury (mTBI). The mTBI model was validated by histopathological and immunohistochemical analyses of brain tissue sections for localization and magnitude of expression of heat-shock protein-70 kDa (HSP70), amyloid precursor protein (APP), cannabinoid receptor-2 (CB2), ionized calcium-binding adapter protein-1 (IBA1), histone deacetylase-4 and -5 (HDAC4 and HDAC5). In comparison to baseline, the expression-activities of HDAC4 and HDAC5 were downregulated in the hippocampus, nucleus accumbens, peri-3rd ventricular part of the thalamus, and substantia nigra at 1-3 days post mTBI, and remained low at 7-8 days post mTBI. Reduced levels of HDAC4 and HDAC5 expression observed in neurons of these brain regions post mTBI were associated with the reduced nuclear and neuropil levels of HDAC4 and HDAC5 with the shift to perinuclear localization of these enzymes. These results support the rationale for the development of therapeutic strategies to upregulate expression-activity of HDACs class IIa post-TBI. PET/CT (MRI) with [18F]TFAHA can facilitate the development and clinical translation of unique therapeutic approaches to upregulate the expression and activity of HDACs class IIa enzymes in the brain after TBI.

Effects of fentanyl on acute locomotor activity, behavioral sensitization, and contextual reward in female and male rats.

BACKGROUND: Although fentanyl has gained widespread prominence, there remains a lack of knowledge on this opioid synthetic agonist, particularly related to sex effects. Therefore, we conducted behavioral tests in female and male rats to measure drug abuse-related responses to fentanyl hypothesizing sex-specific responses. METHODS: Using female and male rats, we measured the effects of acute or repeated administration of fentanyl (20 µg/kg) on locomotor activity (LMA) and behavioral sensitization in an open field test. We further measured contextual-reward and associated locomotor activity during training in a conditioned place preference (CPP) paradigm using a low (4 µg/kg) or high (16 µg/kg) dose of fentanyl. Vaginal lavage samples were collected from female rats in the CPP study, and the estrous phase was determined based on the cytological characterization. RESULTS: Female, but not male, rats showed elevated LMA in response to acute fentanyl and behavioral sensitization to repeated administration of fentanyl. Fentanyl produced significant CPP in both sexes, but it was more potent in males. Finally, our secondary investigation of the estrous cycle on fentanyl-CPP suggests that non-estrus phases, likely reflecting high estradiol, may predict the degree of fentanyl preference in females. CONCLUSIONS: Fentanyl was more potent and/or effective to produce LMA and LMA sensitization in females but more potent to produce CPP in males. Furthermore, the role of sex in fentanyl responses varied across endpoints, and sex differences in LMA were not predictive of sex differences in CPP.

Time course of blast-induced injury in the rat auditory cortex.

Blast exposure is an increasingly significant health hazard and can have a range of debilitating effects, including auditory dysfunction and traumatic brain injury. To assist in the development of effective treatments, a greater understanding of the mechanisms of blast-induced auditory damage and dysfunction, especially in the central nervous system, is critical. To elucidate this area, we subjected rats to a unilateral blast exposure at 22 psi, measured their auditory brainstem responses (ABRs), and histologically processed their brains at 1 day, 1 month, and 3-month survival time points. The left and right auditory cortices was assessed for astrocytic reactivity and axonal degenerative changes using glial fibrillary acidic protein immunoreactivity and a silver impregnation technique, respectively. Although only unilateral hearing loss was induced, astrocytosis was bilaterally elevated at 1 month post-blast exposure compared to shams, and showed a positive trend of elevation at 3 months post-blast. Axonal degeneration, on the other hand, appeared to be more robust at 1 day and 3 months post-blast. Interestingly, while ABR threshold shifts recovered by the 1 and 3-month time-points, a positive correlation was observed between rats' astrocyte counts at 1 month post-blast and their threshold shifts at 1 day post-blast. Taken together, our findings suggest that central auditory damage may have occurred due to biomechanical forces from the blast shockwave, and that different indicators/types of damage may manifest over different timelines.

Neuronal Injury and Glial Changes Are Hallmarks of Open Field Blast Exposure in Swine Frontal Lobe.

With the rapid increase in the number of blast induced traumatic brain injuries and associated neuropsychological consequences in veterans returning from the operations in Iraq and Afghanistan, the need to better understand the neuropathological sequelae following exposure to an open field blast exposure is still critical. Although a large body of experimental studies have attempted to address these pathological changes using shock tube models of blast injury, studies directed at understanding changes in a gyrencephalic brain exposed to a true open field blast are limited and thus forms the focus of this study. Anesthetized, male Yucatan swine were subjected to forward facing medium blast overpressure (peak side on overpressure 224-332 kPa; n = 7) or high blast overpressure (peak side on overpressure 350-403 kPa; n = 5) by detonating 3.6 kg of composition-4 charge. Sham animals (n = 5) were subjected to all the conditions without blast exposure. After a 3-day survival period, the brain was harvested and sections from the frontal lobes were processed for histological assessment of neuronal injury and glial reactivity changes. Significant neuronal injury in the form of beta amyloid precursor protein immunoreactive zones in the gray and white matter was observed in the frontal lobe sections from both the blast exposure groups. A significant increase in the number of astrocytes and microglia was also observed in the blast exposed sections compared to sham sections. We postulate that the observed acute injury changes may progress to chronic periods after blast and may contribute to short and long-term neuronal degeneration and glial mediated inflammation.

Quantitative electroencephalography in a swine model of blast-induced brain injury.

OBJECTIVE: Electroencephalography (EEG) was used to examine brain activity abnormalities earlier after blast exposure using a swine model to develop a qEEG data analysis protocol. METHODS: Anaesthetized swine were exposed to 420-450 Kpa blast overpressure and survived for 3 days after blast. EEG recordings were performed at 15 minutes before the blast and 15 minutes, 30 minutes, 2 hours and 1, 2 and 3 days post-blast using surface recording electrodes and a Biopac 4-channel data acquisition system. Off-line quantitative EEG (qEEG) data analysis was performed to determine qEEG changes. RESULTS: Blast induced qEEG changes earlier after blast exposure, including a decrease of mean amplitude (MAMP), an increase of delta band power, a decrease of alpha band root mean square (RMS) and a decrease of 90% spectral edge frequency (SEF90). CONCLUSIONS: This study demonstrated that qEEG is sensitive for cerebral injury. The changes of qEEG earlier after the blast indicate the potential of utilization of multiple parameters of qEEG for diagnosis of blast-induced brain injury. Early detection of blast induced brain injury will allow early screening and assessment of brain abnormalities in soldiers to enable timely therapeutic intervention.

Biomechanical Responses of the Brain in Swine Subject to Free-Field Blasts.

Blast-induced traumatic brain injury (bTBI) is a signature wound of modern warfare. The current incomplete understanding of its injury mechanism impedes the development of strategies for effective protection of bTBI. Despite a considerable amount of experimental animal studies focused on the evaluation of brain neurotrauma caused by blast exposure, there is very limited knowledge on the biomechanical responses of the gyrenecephalic brain subjected to primary free-field blast waves imposed in vivo. This study aims to evaluate the external and internal mechanical responses of the brain against different levels of blast loading with Yucatan swine in free field. The incident overpressure (IOP) was generated using 3.6 kg of C4 charge placed at three standoff distances from the swine. Five swine were exposed to a total of 19 blasts. The three average peak IOP pressure levels in this study were 148.8, 278.9, and 409.2 kPa as measured by a pencil probe. The duration of the first positive wave was in the range of 2.1-3 ms. Pressure changes in the brain and head kinematics were recorded with intracranial pressure (ICP) sensors, linear accelerometers, and angular rate sensors. The corresponding average peak ICPs were in the range of 79-143, 210-281, and 311-414 kPa designated as low, medium, and high blast level, respectively. Peak head linear accelerations were in the range of 120-412 g. A positive correlation between IOP and its corresponding biomechanical responses of the brain was also observed. These experimental data can be used to validate computer models of bTBI.

Correlation of mechanical impact responses and biomarker levels: A new model for biomarker evaluation in TBI.

A modified Marmarou impact acceleration model was used to help screen biomarkers to assess brain injury severity. Anesthetized male Sprague-Dawley rats were subjected to a closed head injury from 1.25, 1.75 and 2.25 m drop heights. Linear and angular responses of the head were measured in vivo. 24h after impact, cerebrospinal fluid (CSF) and serum were collected. CSF and serum levels of phosphorylated neurofilament heavy (pNF-H), glial fibrillary acidic protein (GFAP), interleukin 6 (IL-6), and amyloid beta (Aß) 1-42 were assessed by enzyme-linked immunosorbent assay (ELISA). Compared to controls, significantly higher CSF and serum pNF-H levels were observed in all impact groups, except between 1.25 m and control in serum. Furthermore, CSF and serum pNF-H levels were significantly different between the impact groups. For GFAP, both CSF and serum levels were significantly higher at 2.25 m compared to 1.75 m, 1.25 m and controls. There was no significant difference in CSF and serum GFAP levels between 1.75 m and 1.25 m, although both groups were significantly higher than control. TBI rats also showed significantly higher levels of IL-6 versus control in both CSF and serum, but no significant difference was observed between each impact group. Levels of Aß were not significantly different between groups. Pearson's correlation analysis showed pNF-H and GFAP levels in CSF and serum had positive correlation with power (rate of impact energy), followed by average linear acceleration and surface righting (p<0.01), which were good predictors for traumatic axonal injury according to histologic assessment in our previous study, suggesting that they are directly related to the injury mechanism. The model used in this study showed a unique ability in elucidating the relationship between biomarker levels and severity of the mechanical trauma to the brain.

Traumatic Brain Injury by a Closed Head Injury Device Induces Cerebral Blood Flow Changes and Microhemorrhages.

OBJECTIVES: Traumatic brain injury is a poly-pathology characterized by changes in the cerebral blood flow, inflammation, diffuse axonal, cellular, and vascular injuries. However, studies related to understanding the temporal changes in the cerebral blood flow following traumatic brain injury extending to sub-acute periods are limited. In addition, knowledge related to microhemorrhages, such as their detection, localization, and temporal progression, is important in the evaluation of traumatic brain injury. MATERIALS AND METHODS: Cerebral blood flow changes and microhemorrhages in male Sprague Dawley rats at 4 h, 24 h, 3 days, and 7 days were assessed following a closed head injury induced by the Marmarou impact acceleration device (2 m height, 450 g brass weight). Cerebral blood flow was measured by arterial spin labeling. Microhemorrhages were assessed by susceptibility-weighted imaging and Prussian blue histology. RESULTS: Traumatic brain injury rats showed reduced regional and global cerebral blood flow at 4 h and 7 days post-injury. Injured rats showed hemorrhagic lesions in the cortex, corpus callosum, hippocampus, and brainstem in susceptibility-weighted imaging. Injured rats also showed Prussian blue reaction products in both the white and gray matter regions up to 7 days after the injury. These lesions were observed in various areas of the cortex, corpus callosum, hippocampus, thalamus, and midbrain. CONCLUSIONS: These results suggest that changes in cerebral blood flow and hemorrhagic lesions can persist for sub-acute periods after the initial traumatic insult in an animal model. In addition, microhemorrhages otherwise not seen by susceptibility-weighted imaging are present in diverse regions of the brain. The combination of altered cerebral blood flow and microhemorrhages can potentially be a source of secondary injury changes following traumatic brain injury and may need to be taken into consideration in the long-term care of these cases.

Acute and subacute effects of the ultrasonic blade and electrosurgery on nerve physiology.

Ultrasonic blades have been shown to cause less acute electrophysiological damage when applied near nerves than monopolar electrosurgery (ES). This study was performed to determine whether the acute nerve damage observed for ES, as well as the relative lack of damage observed for ultrasonic dissection, extends through a subacute timeframe. Muscle incisions were made in rat with the Harmonic(®) Blade (HB) and ES at a distance of 2 mm from the sciatic nerve. Sham surgery was also performed which consisted of similar exposure of the sciatic nerve without use of an energized device. Electrophysiological function was assessed acutely over a 3-h period, and subacutely after a 7-day survival, by monitoring the sciatic nerve compound action potential (CAP), conduction velocity (CV), von Frey hair (VFH) stimulation force, leukocyte infiltration, and impaired axonal transport via ß-amyloid precursor protein (ß-APP) immunocytochemistry. During the acute period, ES produced significantly lower CAP and CV, and higher levels of leukocytes and ß-APP than sham, whereas the ultrasonic blade was not significantly different from sham, and had significantly lower VFH force than ES. After the subacute survival, ES continued to display significantly lower CAP and CV, and higher levels of leukocytes and ß-APP than sham, whereas ultrasonic blade had higher CAP and CV than sham, and lower VFH than ES. This study confirms that incisions made with an ultrasonic blade cause less acute nerve damage than monopolar ES, and are comparable to sham surgery at a distance of 2 mm from the sciatic nerve. The negative effects of electrosurgery extend through at least a 7-day survival period, whereas subacute recovery after application of the ultrasonic blade was comparable to that of sham surgery. For surgical procedures in the vicinity of vital nerves, use of the ultrasonic blade represents a lower risk than ES for both acute and subacute neural trauma.

Blast overpressure induced axonal injury changes in rat brainstem and spinal cord.

INTRODUCTION: Blast induced neurotrauma has been the signature wound in returning soldiers from the ongoing wars in Iraq and Afghanistan. Of importance is understanding the pathomechansim(s) of blast overpressure (OP) induced axonal injury. Although several recent animal models of blast injury indicate the neuronal and axonal injury in various brain regions, animal studies related to axonal injury in the white matter (WM) tracts of cervical spinal cord are limited. OBJECTIVE: The purpose of this study was to assess the extent of axonal injury in WM tracts of cervical spinal cord in male Sprague Dawley rats subjected to a single insult of blast OP. MATERIALS AND METHODS: Sagittal brainstem sections and horizontal cervical spinal cord sections from blast and sham animals were stained by neurofilament light (NF-L) chain and beta amyloid precursor protein immunocytochemistry and observed for axonal injury changes. RESULTS: Observations from this preliminary study demonstrate axonal injury changes in the form of prominent swellings, retraction bulbs, and putative signs of membrane disruptions in the brainstem and cervical spinal cord WM tracts of rats subjected to blast OP. CONCLUSIONS: Prominent axonal injury changes following the blast OP exposure in brainstem and cervical spinal WM tracts underscores the need for careful evaluation of blast induced injury changes and associated symptoms. NF-L immunocytochemistry can be considered as an additional tool to assess the blast OP induced axonal injury.

The effects of ultrasonic and electrosurgery devices on nerve physiology.

BACKGROUND: While the risks associated with the use of electrosurgery near nerves are well known, few studies have examined the neurophysiologic effects of application of the Harmonic Blade, an ultrasonic scalpel, in the vicinity of nerve fibres. This study sought to compare the sub-acute neurophysiologic effects of the Harmonic Blade and electrosurgery after incisions close to the sciatic nerve. METHODS: Incisions were made in rats with the Harmonic Blade and electrosurgery at distances of 1, 2, 3 and 4 mm from the sciatic nerve. Sham surgery was also performed. The compound action potential, conduction velocity and calibrated nylon filament (von Frey hair, VFH) stimulating force were monitored for up to 3 hours after surgery. The sciatic nerve was assessed for inflammation via H&E staining and impaired axonal transport by ß-APP immunohistochemistry. RESULTS: Electrosurgery incisions produced a significantly greater decrease in compound action potential and conduction velocity, and increase in the VFH force than the Harmonic Blade over all time points and distances from the sciatic nerve. The Harmonic Blade was similar to sham surgery for the compound action potential and VFH force. Electrosurgery yielded significantly greater leukocyte infiltration than the Harmonic Blade and produced the highest levels of ß-APP immunoreactive swellings. CONCLUSIONS: Incisions with electrosurgery in the range of 1-4 mm of the sciatic nerve caused substantial changes in neurophysiologic functioning and inflammation. In contrast, the Harmonic Blade was similar to sham surgery in the vicinity of the nerve, producing little observable acute trauma.

Temporal assessment of traumatic axonal injury in the rat corpus callosum and optic chiasm.

Impaired axoplasmic transport (IAT) and neurofilament compaction (NFC), two common axonal pathology processes involved in traumatic axonal injury (TAI), have been well characterized. TAI is found clinically and in animal models in brainstem white matter (WM) tracts and in the corpus callosum (CC), optic chiasm (Och), and internal capsule. Previous published quantitative studies of the time course of TAI expression induced by the Marmarou impact acceleration model have been limited to the brainstem. Accordingly, this study assessed the extent of IAT and NFC in the CC and Och at 8h, 28 h, 3 days and 7 days after traumatic brain injury (TBI) induction by the Marmarou impact acceleration model. IAT peak density was observed at 8h in the CC and 28 h in the Och post-TBI. NFC peak density was observed at 28 h in both structures. The density of IAT and NFC decreased with increasing survival time in both structures. The NFC density time profile followed a similar trend in both the Och and CC, whereas the IAT density time profile was variable between the Och and CC. Furthermore, a strong linear relationship was observed between IAT and NFC in the CC but not in the Och. These findings highlight the heterogeneity of TAI as evidenced by variable IAT and NFC injury time profiles in each anatomical structure. This variability indicates the requirement of multiple markers for a comprehensive TAI evaluation and multiple targeted treatments for TAI polypathology within its therapeutic window time frame.

Impaired axoplasmic transport is the dominant injury induced by an impact acceleration injury device: an analysis of traumatic axonal injury in pyramidal tract and corpus callosum of rats.

Traumatic axonal injury (TAI) involves neurofilament compaction (NFC) and impaired axoplasmic transport (IAT) in distinct populations of axons. Previous quantification studies of TAI focused on limited areas of pyramidal tract (Py) but not its entire length. Quantification of TAI in corpus callosum (CC) and its comparison to that in Py is also lacking. This study assessed and compared the extent of TAI in the entire Py and CC of rats following TBI. TBI was induced by a modified Marmarou impact acceleration device in 31 adult male Sprague Dawley rats by dropping a 450 gram impactor from either 1.25 m or 2.25 m. Twenty-four hours after TBI, TAI was assessed by beta amyloid precursor protein (ß-APP-IAT) and RMO14 (NFC) immunocytochemistry. TAI density (ß-APP and RMO14 axonal swellings, retraction balls and axonal profiles) was counted from panoramic images of CC and Py. Significantly high TAI was observed in 2.25 m impacted rats. ß-APP immunoreactive axons were significantly higher in number than RMO14 immunoreactive axons in both the structures. TAI density in Py was significantly higher than in CC. Based on our parallel biomechanical studies, it is inferred that TAI in CC may be related to compressive strains and that in Py may be related to tensile strains. Overall, IAT appears to be the dominant injury type induced by this model and injury in Py predominates that in CC.

Innervation of cervical ventral facet joint capsule: Histological evidence.

AIM: To assess the presence of nerves in ventral facet joint capsules as facet capsules are generally implicated in neck pain. METHODS: Twenty-four ventral cervical facet joint capsules were harvested from 3 unembalmed cadavers. Paraffin sections from these capsules were processed to identify neurofilament and substance P immunoreactive fibers. Nerve fiber presence was also verified by a silver impregnation method. RESULTS: Neurofilament reactive fibers were observed in sections from 9 capsules. They were observed in areas with collagen fibers and areas with irregular connective tissue. Substance P reactive nerve fibers were found in sections from 7 capsules in similar areas. Silver impregnation also revealed the presence of nerve fibers. The nerve fibers were also found as bundles in the lateral margins of the capsule. A Pacinian corpuscle-like ending was also observed in one specimen. CONCLUSION: Nerve fibers revealed by neurofilament immunoreactivity and silver staining support innervation of the ventral aspect of the facet joint capsule. The presence of substance P reactive fibers supports the potential role of these elements in mediating pain. The presence of a Pacinian-like ending implicates a potential role in joint movement.

Quantitative relationship between axonal injury and mechanical response in a rodent head impact acceleration model.

A modified Marmarou impact acceleration model was developed to study the mechanical responses induced by this model and their correlation to traumatic axonal injury (TAI). Traumatic brain injury (TBI) was induced in 31 anesthetized male Sprague-Dawley rats (392±13 g) by a custom-made 450-g impactor from heights of 1.25 m or 2.25 m. An accelerometer and angular rate sensor measured the linear and angular responses of the head, while the impact event was captured by a high-speed video camera. TAI distribution along the rostro-caudal direction, as well as across the left and right hemispheres, was determined using ß-amyloid precursor protein (ß-APP) immunocytochemistry, and detailed TAI injury maps were constructed for the entire corpus callosum. Peak linear acceleration 1.25 m and 2.25 m impacts were 666±165 g and 907±501 g, respectively. Peak angular velocities were 95±24 rad/sec and 124±48 rad/sec, respectively. Compared to the 2.25-m group, the observed TAI counts in the 1.25-m impact group were significantly lower. Average linear acceleration, peak angular velocity, average angular acceleration, and surface righting time were also significantly different between the two groups. A positive correlation was observed between normalized total TAI counts and average linear acceleration (R(2)=0.612, p<0.05), and time to surface right (R(2)=0.545, p<0.05). Our study suggested that a 2.25-m drop in the Marmarou model may not always result in a severe injury, and TAI level is related to the linear and angular acceleration response of the rat head during impact, not necessarily the drop height.

Endothelin receptor A antagonism reduces the extent of diffuse axonal injury in a rodent model of traumatic brain injury.

OBJECTIVES: While endothelin-1 and its receptors have traditionally been associated with mediating vasoreactivity, we have recently shown that the vast majority of endothelin receptor A expression following traumatic brain injury is localized within the neuron. While it has been suggested that endothelin receptor A plays a role in influencing neuronal integrity, the significance of neuronally expressed endothelin receptor A remains unclear. One report suggests that endothelin-1 signaling mediates diffuse axonal injury. Therefore, this work sought to determine whether treatment with BQ-123, a selective endothelin receptor A antagonist, diminishes the extent of diffuse axonal injury following trauma. METHODS: A total of 12 male Sprague-Dawley rats (350-400 g) were used in this study. Two groups (n = 6 per group) were generated as follows: sham operation and traumatic brain injury+1·0 mg/kg BQ-123 delivered intravenously 30 minutes prior to the injury. Trauma was induced using a weight acceleration impact device. Animals were terminated 24 or 48 hours after trauma, and a series of six coronal sections through the entire anterior-posterior extent of the corpus callosum were selected from each brain for quantification of diffuse axonal injury by beta-amyloid precursor protein immunostaining. RESULTS: Our data indicated that animals treated with BQ-123 30 minutes prior to trauma showed a significant reduction in diffuse axonal injury in corpus callosum at both 24 and 48 hours post-injury. CONCLUSION: The results show that endothelin receptor A antagonism reduced the extent of diffuse axonal injury, demonstrating a potential influence of the endothelin system on the intra-axonal cascade of molecular events underlying diffuse axonal injury.

Injury predictors for traumatic axonal injury in a rodent head impact acceleration model.

A modified Marmarou impact acceleration injury model was developed to study the kinematics of the rat head to quantify traumatic axonal injury (TAI) in the corpus callosum (CC) and brainstem pyramidal tract (Py), to determine injury predictors and to establish injury thresholds for severe TAI. Thirty-one anesthetized male Sprague-Dawley rats (392±13 grams) were impacted using a modified impact acceleration injury device from 2.25 m and 1.25 m heights. Beta-amyloid precursor protein (ß-APP) immunocytochemistry was used to assess and quantify axonal changes in CC and Py. Over 600 injury maps in CC and Py were constructed in the 31 impacted rats. TAI distribution along the rostro-caudal direction in CC and Py was determined. Linear and angular responses of the rat head were monitored and measured in vivo with an attached accelerometer and angular rate sensor, and were correlated to TAI data. Logistic regression analysis suggested that the occurrence of severe TAI in CC was best predicted by average linear acceleration, followed by power and time to surface righting. The combination of average linear acceleration and time to surface righting showed an improved predictive result. In Py, severe TAI was best predicted by time to surface righting, followed by peak and average angular velocity. When both CC and Py were combined, power was the best predictor, and the combined average linear acceleration and average angular velocity was also found to have good injury predictive ability. Receiver operator characteristic curves were used to assess the predictive power of individual and paired injury predictors. TAI tolerance curves were also proposed in this study.

Muscular response to physiologic tensile stretch of the caprine c5/6 facet joint capsule: dynamic recruitment thresholds and latencies.

This study examined the cervical muscle response to physiologic, high-rate (100 mm/s) tensile facet joint capsule (FJC) stretch. Six in-vivo caprine C5/6 FJC preparations were subjected to an incremental tensile loading paradigm. EMG activity was recorded from the right trapezius (TR) and multifidus (MF) muscle groups at the C5 and C6 levels; and from the sternomastoid (SM) and longus colli (LC) muscle groups bilaterally at the C5/6 level; during FJC stretch. Capsule load during the displacement applications was recorded via a miniature load cell, and 3D capsule strains (based on stereoimaging of an array of markers on the capsule surface) were reconstructed using finite element methods. EMG traces from each muscle were examined for onset of muscular activity. Capsule strains and loads at the time of EMG onset were recorded for each muscle, as was the time from the onset of FJC stretch to the onset of muscle activity. All muscles were responsive to physiologic high-rate FJC stretch. The deep muscles (MF and LC) were recruited at significantly smaller capsule loads and onset latencies than the superficial muscles (TR and SM). MF activation strain was significantly smaller than LC and TR activation strains. These data were also compared to previously published low-rate data. MF was the first muscle group to be recruited regardless of the activation criterion under consideration (i.e. strain, load, or latency) or the rate of FJC stretch. LC recruitment occurred significantly sooner under high-rate vs. low-rate FJC stretch. The results of this study provide further evidence of extensive ligamento-muscular reflex pathways between the FJC and the cervical musculature, which are responsive to both low-rate and high-rate FJC stretch. These data add to our knowledge of the dynamic response of paraspinal muscles relative to facet joint motion and provide a unique contribution to enhance the precision of computer-simulated impacts.

Structural and functional changes in nerve roots due to tension at various strains and strain rates: an in-vivo study.

This study investigates the functional and structural responses of spinal nerve roots in vivo to various strains and strain rates. Seventy-two L5 dorsal nerve roots from male Sprague-Dawley rats were each subjected to a predetermined strain (<10%, 10-20%, and >20%; n = 8) and rate (0.01 mm/sec, 1 mm/sec, or 15 mm/sec; n = 24). Neurophysiologic recordings were performed before and after stretch to determine changes in conduction velocity (CV), amplitude, and area of the compound action potential (CAP). Morphological injury as evident by primary and secondary axotomy as well as impaired axoplasmic transport (IAT) was determined using the palmgren silver impregnation technique and betaAPP immunostaining, respectively. The results from neurophysiologic recordings indicate that as strain and rate increased, there was a decrease in CV, amplitude, and area of the CAP. Further, high strains led to a complete conduction block that appeared to be rate dependent. Strains of 16%, 10%, and 9%, at 0.01 mm/sec, 1 mm/sec, and 15 mm/sec, respectively, led to 50% probability of complete conduction block in the nerve roots. Results from histological assessment indicate an increase in periaxonal spacing (secondary axotomy) and torn fibers (primary axotomy), as well as impaired IAT, with increasing strain and rate. Overall, the results from the current study indicate that (1) functional nerve root injuries as evident by changes in the CV, amplitude, and area of the CAP are strain- and rate-dependent; (2) high strains at low rates cause complete conduction block in the roots, while a similar block was observed at lower strains at the high rate; (3) the extent of IAT and primary and secondary axotomy occurred concomitant with functional injury and were strain- and rate-dependent.