Whole-body Vibration at Thoracic Resonance Induces Sustained Pain and Widespread Cervical Neuroinflammation in the Rat.

BACKGROUND: Whole-body vibration (WBV) is associated with back and neck pain in military personnel and civilians. However, the role of vibration frequency and the physiological mechanisms involved in pain symptoms are unknown. QUESTIONS/PURPOSES: This study asked the following questions: (1) What is the resonance frequency of the rat spine for WBV along the spinal axis, and how does frequency of WBV alter the extent of spinal compression/extension? (2) Does a single WBV exposure at resonance induce pain that is sustained? (3) Does WBV at resonance alter the protein kinase C epsilon (PKCε) response in the dorsal root ganglia (DRG)? (4) Does WBV at resonance alter expression of calcitonin gene-related peptide (CGRP) in the spinal dorsal horn? (5) Does WBV at resonance alter the spinal neuroimmune responses that regulate pain? METHODS: Resonance of the rat (410 ± 34 g, n = 9) was measured by imposing WBV at frequencies from 3 to 15 Hz. Separate groups (317 ± 20 g, n = 10/treatment) underwent WBV at resonance (8 Hz) or at a nonresonant frequency (15 Hz). Behavioral sensitivity was assessed throughout to measure pain, and PKCε in the DRG was quantified as well as spinal CGRP, glial activation, and cytokine levels at Day 14. RESULTS: Accelerometer-based thoracic transmissibility peaks at 8 Hz (1.86 ± 0.19) and 9 Hz (1.95 ± 0.19, mean difference [MD] 0.290 ± 0.266, p < 0.03), whereas the video-based thoracic transmissibility peaks at 8 Hz (1.90 ± 0.27), 9 Hz (2.07 ± 0.20), and 10 Hz (1.80 ± 0.25, MD 0.359 ± 0.284, p < 0.01). WBV at 8 Hz produces more cervical extension (0.745 ± 0.582 mm, MD 0.242 ± 0.214, p < 0.03) and compression (0.870 ± 0.676 mm, MD 0.326 ± 0.261, p < 0.02) than 15 Hz (extension, 0.503 ± 0.279 mm; compression, 0.544 ± 0.400 mm). Pain is longer lasting (through Day 14) and more robust (p < 0.01) after WBV at the resonant frequency (8 Hz) compared with 15 Hz WBV. PKCε in the nociceptors of the DRG increases according to the severity of WBV with greatest increases after 8 Hz WBV (p < 0.03). However, spinal CGRP, cytokines, and glial activation are only evident after painful WBV at resonance. CONCLUSIONS: WBV at resonance produces long-lasting pain and widespread activation of a host of nociceptive and neuroimmune responses as compared with WBV at a nonresonance condition. Based on this work, future investigations into the temporal and regional neuroimmune response to resonant WBV in both genders would be useful. CLINICAL RELEVANCE: Although WBV is a major issue affecting the military population, there is little insight about its mechanisms of injury and pain. The neuroimmune responses produced by WBV are similar to other pain states, suggesting that pain from WBV may be mediated by similar mechanisms as other neuropathic pain conditions. This mechanistic insight suggests WBV-induced injury and pain may be tempered by antiinflammatory intervention.

Relevant Anatomic and Morphological Measurements of the Rat Spine: Considerations for Rodent Models of Human Spine Trauma.

STUDY DESIGN: Basic science study measuring anatomical features of the cervical and lumbar spine in rat with normalized comparison with the human. OBJECTIVE: The goal of this study is to comprehensively compare the rat and human cervical and lumbar spines to investigate whether the rat is an appropriate model for spine biomechanics investigations. SUMMARY OF BACKGROUND DATA: Animal models have been used for a long time to investigate the effects of trauma, degenerative changes, and mechanical loading on the structure and function of the spine. Comparative studies have reported some mechanical properties and/or anatomical dimensions of the spine to be similar between various species. However, those studies are largely limited to the lumbar spine, and a comprehensive comparison of the rat and human spines is lacking. METHODS: Spines were harvested from male Holtzman rats (n = 5) and were scanned using micro- computed tomography and digitally rendered in 3 dimensions to quantify the spinal bony anatomy, including the lateral width and anteroposterior depth of the vertebra, vertebral body, and spinal canal, as well as the vertebral body and intervertebral disc heights. Normalized measurements of the vertebra, vertebral body, and spinal canal of the rat were computed and compared with corresponding measurements from the literature for the human in the cervical and lumbar spinal regions. RESULTS: The vertebral dimensions of the rat spine vary more between spinal levels than in humans. Rat vertebrae are more slender than human vertebrae, but the width-to-depth axial aspect ratios are very similar in both species in both the cervical and lumbar regions, especially for the spinal canal. CONCLUSION: The similar spinal morphology in the axial plane between rats and humans supports using the rat spine as an appropriate surrogate for modeling axial and shear loading of the human spine.

Upregulation of BDNF and NGF in cervical intervertebral discs exposed to painful whole-body vibration.

STUDY DESIGN: In vivo study defining expression of the neurotrophins, brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF), in cervical intervertebral discs after painful whole-body vibration (WBV). OBJECTIVE: The goal of this study is to determine if BDNF and NGF are expressed in cervical discs after painful WBV in a rat model. SUMMARY OF BACKGROUND DATA: WBV is a possible source of neck pain and has been implicated as increasing the risk for disc disorders. Typically, aneural regions of painful human lumbar discs exhibit hyperinnervation, suggesting nerve ingrowth as potentially contributing to disc degeneration and pain. BDNF and NGF are upregulated in painfully degenerate lumbar discs and hypothesized to contribute to this pathology. METHODS: Male Holtzman rats underwent 7 days of repeated WBV (15 Hz, 30 min/d) or sham exposures, followed by 7 days of rest. Cervical discs were collected for analysis of BDNF and NGF expression through RT-qPCR and Western blot analysis. Immunohistochemistry also evaluated their regional expression in the disc. RESULTS: Vibration significantly increases BDNF messenger ribonucleic acid (mRNA) levels (P=0.036), as well as total-NGF mRNA (P=0.035). Protein expression of both BDNF (P=0.006) and the 75-kDa NGF (P=0.045) increase by nearly 4- and 10-fold, respectively. Both BDNF mRNA (R=0.396; P=0.012) and protein (R=0.280; P=0.035) levels are significantly correlated with the degree of behavioral sensitivity (i.e., pain) at day 14. Total-NGF mRNA is also significantly correlated with the extent of behavioral sensitivity (R=0.276; P=0.044). Both neurotrophins are most increased in the inner annulus fibrosus and nucleus pulposus. CONCLUSION: The increases in BDNF and NGF in the cervical discs after painful vibration are observed in typically aneural regions of the disc, consistent with reports of its hyperinnervation. Yet, the induction of nerve ingrowth into the disc was not explicitly investigated. Neurotrophin expression also correlates with behavioral sensitivity, suggesting a role for both neurotrophins in the development of disc pain. LEVEL OF EVIDENCE: N/A.

Whole body vibration induces forepaw and hind paw behavioral sensitivity in the rat.

Whole body vibration (WBV) has been linked to neck and back pain, but the biomechanical and physiological mechanisms responsible for its development and maintenance are unknown. A rodent model of WBV was developed in which rats were exposed to different WBV paradigms, either daily for 7 consecutive days (repeated WBV) or two single exposures at Day 0 and 7 (intermittent WBV). Each WBV session lasted for 30 min and was imposed at a frequency of 15 Hz and RMS platform acceleration of 0.56 ± 0.07 g. Changes in the withdrawal response of the forepaw and hind paw were measured, and were used to characterize the onset and maintenance of behavioral sensitivity. Accelerations and displacements of the rat and deformations in the cervical and lumbar spines were measured during WBV to provide mechanical context for the exposures. A decrease in withdrawal threshold was induced at 1 day after the first exposure in both the hind paw and forepaw. Repeated WBV exhibited a sustained reduction in withdrawal threshold in both paws and intermittent WBV induced a sustained response only in the forepaw. Cervical deformations were significantly elevated which may explain the more robust forepaw response. Findings suggest that a WBV exposure leads to behavioral sensitivity.

ProDisc cervical arthroplasty does not alter facet joint contact pressure during lateral bending or axial torsion.

STUDY DESIGN: A biomechanical study of facet joint pressure after total disc replacement using cadaveric human cervical spines during lateral bending and axial torsion. OBJECTIVE: The goal was to measure the contact pressure in the facet joint in cadaveric human cervical spines subjected to physiologic lateral bending and axial torsion before and after implantation of a ProDisc-C implant. SUMMARY OF BACKGROUND DATA: Changes in facet biomechanics can damage the articular cartilage in the joint, potentially leading to degeneration and painful arthritis. Few cadaveric and computational studies have evaluated the changes in facet joint loading during spinal loading with an artificial disc implanted. Computational models have predicted that the design and placement of the implant influence facet joint loading, but limited cadaveric studies document changes in facet forces and pressures during nonsagittal bending after implantation of a ProDisc. As such, little is known about the local facet joint mechanics for these complicated loading scenarios in the cervical spine. METHODS: Seven osteoligamentous C2-T1 cadaveric cervical spines were instrumented with a transducer to measure the C5-C6 facet pressure profiles during physiological lateral bending and axial torsion, before and after implantation of a ProDisc-C at that level. Rotations at that level and global cervical spine motions and loads were also quantified. RESULT.: Global and segmental rotations were not altered by the disc implantation. Facet contact pressure increased after implantation during ipsilateral lateral bending and contralateral torsion, but that increase was not significant compared with the intact condition. CONCLUSION: Implantation of a ProDisc-C does not significantly modify the kinematics and facet pressure at the index level in cadaveric specimens during lateral bending and axial torsion. However, changes in facet contact pressures after disc arthroplasty may have long-term effects on spinal loading and cartilage degeneration and should be monitored in vivo.

Facet joint contact pressure is not significantly affected by ProDisc cervical disc arthroplasty in sagittal bending: a single-level cadaveric study.

BACKGROUND CONTEXT: Total disc arthroplasty is a motion-preserving spinal procedure that has been investigated for its impact on spinal motions and adjacent-level degeneration. However, the effects of disc arthroplasty on facet joint biomechanics remain undefined despite the critical role of these posterior elements on guiding and limiting spinal motion. PURPOSE: The goal was to measure the pressure in the facet joint in cadaveric human cervical spines subjected to sagittal bending before and after implantation of the ProDisc-C (Synthes Spine Company, L.P, West Chester, PA, USA). STUDY DESIGN: A biomechanical study was performed using cadaveric human cervical spines during sagittal bending in the intact and implanted conditions. METHODS: Seven C2-T1 osteoligamentous cadaveric cervical spines were instrumented with a transducer to measure the C5-C6 facet pressure profiles during physiological sagittal bending, before and after implantation of a ProDisc-C at that level. Rotations of the index segment and global cervical spine were also quantified. RESULTS: The mean C5-C6 range of motion significantly increased (p=.009) from 9.6°±5.1° in the intact condition to 16.2°±3.6° after implantation. However, despite such changes in rotation, there was no significant difference in the facet contact pressure during extension between the intact (64±30 kPa) and implanted (44±55 kPa) conditions. Similarly, there was no difference in facet pressure developed during flexion. CONCLUSIONS: Although implantation of a ProDisc-C arthroplasty device at the C5-C6 level increases angular rotations, it does not significantly alter the local facet pressure at the index level in flexion or extension. Using a technique that preserves the capsular ligament, this study provides the first direct measurement of cervical facet pressure in a disc arthroplasty condition.

The potential for salmon fibrin and thrombin to mitigate pain subsequent to cervical nerve root injury.

Nerve root compression is a common cause of radiculopathy and induces persistent pain. Mammalian fibrin is used clinically as a coagulant but presents a variety of risks. Fish fibrin is a potential biomaterial for neural injury treatment because it promotes neurite outgrowth, is non-toxic, and clots readily at lower temperatures. This study administered salmon fibrin and thrombin following nerve root compression and measured behavioral sensitivity and glial activation in a rat pain model. Fibrin and thrombin each significantly reduced mechanical allodynia compared to injury alone (p < 0.02). Painful compression with fibrin exhibited allodynia that was not different from sham for any day using stimulation by a 2 g filament; allodynia was only significantly different (p < 0.043) from sham using the 4 g filament on days 1 and 3. By day 5, responses for fibrin treatment decreased to sham levels. Allodynia following compression with thrombin treatment were unchanged from sham at any time point. Macrophage infiltration at the nerve root and spinal microglial activation were only mildly modified by salmon treatments. Spinal astrocytic expression decreased significantly with fibrin (p < 0.0001) but was unchanged from injury responses for thrombin treatment. Results suggest that salmon fibrin and thrombin may be suitable biomaterials to mitigate pain.

Contact pressure in the facet joint during sagittal bending of the cadaveric cervical spine.

The facet joint contributes to the normal biomechanical function of the spine by transmitting loads and limiting motions via articular contact. However, little is known about the contact pressure response for this joint. Such information can provide a quantitative measure of the facet joint's local environment. The objective of this study was to measure facet pressure during physiologic bending in the cervical spine, using a joint capsule-sparing technique. Flexion and extension bending moments were applied to six human cadaveric cervical spines. Global motions (C2-T1) were defined using infra-red cameras to track markers on each vertebra. Contact pressure in the C5-C6 facet was also measured using a tip-mounted pressure transducer inserted into the joint space through a hole in the postero-inferior region of the C5 lateral mass. Facet contact pressure increased by 67.6 ± 26.9 kPa under a 2.4 Nm extension moment and decreased by 10.3 ± 9.7 kPa under a 2.7 Nm flexion moment. The mean rotation of the overall cervical specimen motion segments was 9.6 ± 0.8° and was 1.6 ± 0.7° for the C5-C6 joint, respectively, for extension. The change in pressure during extension was linearly related to both the change in moment (51.4 ± 42.6 kPa/Nm) and the change in C5-C6 angle (18.0 ± 108.9 kPa/deg). Contact pressure in the inferior region of the cervical facet joint increases during extension as the articular surfaces come in contact, and decreases in flexion as the joint opens, similar to reports in the lumbar spine despite the difference in facet orientation in those spinal regions. Joint contact pressure is linearly related to both sagittal moment and spinal rotation. Cartilage degeneration and the presence of meniscoids may account for the variation in the pressure profiles measured during physiologic sagittal bending. This study shows that cervical facet contact pressure can be directly measured with minimal disruption to the joint and is the first to provide local pressure values for the cervical joint in a cadaveric model.

Metabotropic glutamate receptor-5 and protein kinase C-epsilon increase in dorsal root ganglion neurons and spinal glial activation in an adolescent rat model of painful neck injury.

There is growing evidence that neck pain is common in adolescence and is a risk factor for the development of chronic neck pain in adulthood. The cervical facet joint and its capsular ligament is a common source of pain in the neck in adults, but its role in adolescent pain remains unknown. The aim of this study was to define the biomechanics, behavioral sensitivity, and indicators of neuronal and glial activation in an adolescent model of mechanical facet joint injury. A bilateral C6-C7 facet joint distraction was imposed in an adolescent rat and biomechanical metrics were measured during injury. Following injury, forepaw mechanical hyperalgesia was measured, and protein kinase C-epsilon (PKCɛ) and metabotropic glutamate receptor-5 (mGluR5) expression in the dorsal root ganglion and markers of spinal glial activation were assessed. Joint distraction induced significant mechanical hyperalgesia during the 7 days post-injury (p < 0.001). Painful injury significantly increased PKCɛ expression in small- and medium-diameter neurons compared to sham (p < 0.05) and naïve tissue (p < 0.001). Similarly, mGluR5 expression was significantly elevated in small-diameter neurons after injury (p < 0.05). Spinal astrocytic activation after injury was also elevated over sham (p < 0.035) and naïve (p < 0.0001) levels; microglial activation was only greater than naïve levels (p < 0.006). Mean strains in the facet capsule during injury were 32.8 ± 12.9%, which were consistent with the strains associated with comparable degrees of hypersensitivity in the adult rat. These results suggest that adolescents may have a lower tissue tolerance to induce pain and associated nociceptive response than do adults.

Spinal microglial proliferation is evident in a rat model of painful disc herniation both in the presence of behavioral hypersensitivity and following minocycline treatment sufficient to attenuate allodynia.

Although spinal glia acquire a reactive profile in radiculopathy, glial cell proliferation remains largely unstudied. This study investigated spinal glial proliferation in a model simulating painful disc herniation; the C7 nerve root underwent compression and chromic gut suture exposure or sham procedures. A subset of injured rats received minocycline injections prior to injury. Allodynia was assessed and bromodeoxyuridine (BrdU) was injected 2 hr before tissue harvest on day 1 or 3. Spinal cell proliferation and phenotype identification were assayed by fluorescent colabeling with antibodies to BrdU and either glial fibrillary acidic protein (astrocytes) or Iba1 (microglia). At day 1, ipsilateral allodynia was significantly increased (P < 0.001) for injury over sham. Minocycline treatment significantly decreased ipsilateral allodynia to sham levels at day 1 (P < 0.001). At day 3, ipsilateral allodynia remained and contralateral allodynia was also present for injury (P< 0.003) over sham. The number of BrdU-positive cells in the ipsilateral spinal dorsal horn at day 1 after injury was significantly elevated (P < 0.001) over sham. Approximately 70% of BrdU-positive cells labeled positively for Iba1; dividing microglia were significantly increased (P < 0.004) in the ipsilateral dorsal horn at day 1 following injury compared with sham. Spinal cellular proliferation after injury was not changed by minocycline injection. By day 3, the number of BrdU-positive cells had returned to sham levels bilaterally. Data indicate that spinal microglia proliferate after injury but that proliferation is not abolished by minocycline treatment that attenuates allodynia, indicating that spinal microglial proliferation may be related to injury and may not be linked to changes in sensory perception.