Spinal Cord Injury In Dogs Research Articles

Outcome in paraplegic dogs with or without pain perception due to thoracolumbar fibrocartilaginous embolic myelopathy or acute non-compressive nucleus pulposus extrusion.

Fibrocartilaginous embolic myelopathy (FCEM) and acute non-compressive nucleus pulposus extrusion (ANNPE) are common causes of acute spinal cord injury in dogs. Outcome among paraplegic deep pain positive (DPP) and deep pain negative (DPN) dogs with either condition and factors influencing recovery have not been clearly established. Dogs with thoracolumbar FCEM or ANNPE resulting in paraplegia presenting to university hospitals between 2012 and 2022 were retrospectively included. Diagnosis of FCEM or ANNPE was based on clinical and magnetic resonance imaging findings. Outcome was defined as successful (recovery of independent ambulation) or unsuccessful (non-ambulatory ≥3 months following diagnosis or at the time of death/euthanasia). Logistic regression analysis was performed to investigate associations between clinical or imaging variables and outcome. Thirty-one dogs were included. In total, 14 dogs were initially paraplegic DPP (8 FCEM, 6 ANNPE) and 17 dogs were paraplegic DPN (11 FCEM, 6 ANNPE). Outcome was available for 26 dogs (14 DPP, 12 DPN) with a median follow-up time of 182 days (range 0-2,311) including 2 dogs euthanized at the time of diagnosis; 1 of 12 DPN dogs (8.3%) regained independent ambulation, whereas 9 of 14 DPP dogs (64.3%) regained independent ambulation. DPN dogs had a significantly higher risk of not regaining independent ambulation compared with DPP dogs (OR: 47.40, 95% CI: 2.09-1073.99). No other variables were associated with outcome. While recovery of ambulation was possible, these results confirm that the absence of pain perception is a useful negative prognostic indicator in dogs with severe thoracolumbar FCEM or ANNPE.

Balloon compression-induced spinal cord injury in canines: a large animal model for spinal cord injury research.

Spinal cord injury (SCI) is a life-altering condition that severely impacts an individual's functional capabilities and has significant implications for both the individual and society. Large animal models are crucial for understanding the pathology and biomechanics of SCI. Dogs (Canis lupus familiaris) are promising models for SCI research due to their anatomical and histopathological similarities to humans. Balloon compression is an established method for inducing controlled SCI in canines. In this study, we optimized a balloon compression procedure for inducing SCI in dogs, aiming to develop a reliable model for future in vivo studies. Our methodology successfully induced total motoric loss in canines, observed for seven days, a critical period for therapeutic interventions. Histopathological examinations using Luxol fast blue (LFB) staining revealed total demyelination in intralesional samples, confirming the structural damage caused by balloon compression. We concluded that a balloon compression model at the T10-T11 vertebral level, with an inflated balloon volume of 1.0 ml, induced SCI while minimizing the risk of balloon rupture. Longer duration of compression ensures total paralysis in this model, providing a platform for testing therapeutic interventions during the acute phase of SCI. The canine model generated consistent data and facilitated straightforward observational findings.

Randomized Controlled Trial of Durotomy as an Adjunct to Routine Decompressive Surgery for Dogs With Severe Acute Spinal Cord Injury.

Although many interventions for acute spinal cord injury (SCI) appear promising in experimental models, translation directly from experimental animals to human patients is a large step that can be problematic. Acute SCI occurs frequently in companion dogs and may provide a model to ease translation. Recently, incision of the dura has been highlighted in both research animals and human patients as a means of reducing intraspinal pressure, with a view to improving perfusion of the injured tissue and enhancing functional recovery. Observational clinical data in humans and dogs support the notion that it may also improve functional outcome. Here, we report the results of a multi-center randomized controlled trial of durotomy as an adjunct to traditional decompressive surgery for treatment of severe thoracolumbar SCI caused by acute intervertebral disc herniation in dogs. Sample-size calculation was based on the proportion of dogs recovering ambulation improving from an expected 55% in the traditional surgery group to 70% in the durotomy group. Over a 3.5-year period, we enrolled 140 dogs, of which 128 had appropriate duration of follow-up. Overall, 65 (51%) dogs recovered ambulation. Recovery in the traditional decompression group was 35 of 62 (56%) dogs, and in the durotomy group 30 of 66 (45%) dogs, associated with an odds ratio of 0.643 (95% confidence interval: 0.320-1.292) and z-score of -1.24. This z-score indicates trial futility to reach the target 15% improvement over traditional surgery, and the trial was terminated at this stage. We conclude that durotomy is ineffective in improving functional outcome for severe acute thoracolumbar SCI in dogs. In the future, these data can be compared with similar data from clinical trials on duraplasty in human patients and will aid in determining the predictive validity of the "companion dog model" of acute SCI.

Quantification of spinal ataxia in dogs with thoracolumbar spinal cord injury.

The clinical sign of ataxia is related to several neurological diseases and is seen in conjunction with paresis in dogs with spinal cord injury (SCI). Endeavours to objectify canine spinal ataxia in SCI remain limited. The aim of this clinical study was to determine and quantify differences between gait characteristics of ataxic dogs with thoracolumbar myelopathy and healthy control dogs using a computer-and treadmill-based gait analysis system. Five dogs with spinal ataxia and six healthy dogs underwent video-and computer-assisted gait analysis while walking on a four-ground reaction force plate treadmill system (maximum speed of 0.7 m/s). Spatio-temporal and kinetic gait characteristics regarding the dogs' locomotion were analysed with a focus on the individual coefficient of variation (CV), as a potential measure for quantification of the level of ataxia. Ataxic dogs with thoracolumbar SCI showed no effect on symmetry indices but higher variability in spatio-temporal and kinetic gait parameters mainly in the pelvic, but also in the thoracic limbs. Double support phase of the individual limb was prolonged in SCI dogs at the cost of the single support and swing phase. Reduced peaks of ground reaction forces (GRF) could potentially be explained by reduction of muscle strength, as a strategy of avoiding falling by taking enthusiastic steps, or by alteration of the rhythmogenic spinal circuits between the pelvic and thoracic limb pattern generators.

A 50-step walking test for analysis of recovery after decompressive surgery for thoracolumbar disc herniation in dogs.

BackgroundDespite its importance, there is no agreed definition of recovery of ambulation in dogs with spinal cord injury.ObjectivesTo validate a new walking test in dogs recovering from thoracolumbar spinal cord injury.AnimalsTwo hundred twenty‐four dogs weighing <20 kg: 120 normally ambulatory dogs, plus 104 dogs undergoing decompressive surgery for acute thoracolumbar intervertebral disc herniation.MethodsProspective cohort studies. The distance each freely‐ambulatory dog walked during 50 step cycles was regressed on ulna length. For each postsurgical dog, we recorded when the calculated 50‐step distance was completed without falling, or their inability to complete this distance by 4 months or more after surgery. Bayesian analysis compared outcomes for presurgical neurologic categories; association of recovery with several preoperative variables was explored using logistic and time‐to‐event regression.ResultsFor control dogs, 50‐step distance (m) = 1.384 × ulnar length (cm) + 2.773. In postsurgical dogs, the 50‐step test provided decisive evidence that deep pain‐negative dogs were less likely to recover ambulation than dogs with intact pain perception (12/29 recovered vs 71/75; Bayes factor [BF] = 5.9 × 106) and, if they did recover, it took much longer (median 91 days vs median 14 days; BF = 1.5 × 103). Exploratory analysis suggested that presurgical neurologic status (subhazard ratio [SHR] = 0.022; P < .001) and duration of presurgical anesthesia (SHR = 0.740; P = .04) were associated with rapidity of recovery.Conclusions and Clinical ImportanceThis straightforward 50‐step walking test provides robust data on ambulatory recovery well‐suited to large scale pragmatic trials on treatment of thoracolumbar spinal cord injury in dogs.

Application of mesenchymal stem cells for treating spinal cord injury in dogs: Mechanisms and their therapeutic efficacy

Despite progress in the treatment of spinal cord injury (SCI), recovery of the spinal cord with normal motor andsensory activities remains a challenge due to the complex anatomy of the spine, and its limited regeneration potential in mammals. Recently, the clinical application of mesenchymal stem cells (MSCs) in SCI led to promising results in both human and veterinary medicine. The mechanism by which MSCs might promote wound healing of SCI has been extensively investigated. Previous reports have suggested that transplanted MSCs enhance the numbers of neurons and glial cells, prevent neuronal apoptosis, inhibit inflammation, stimulate vascular angiogenesis, axonal myelination, and neuro-regeneration at the lesion site. It has also been reported that therapeutic applicability depends on the source of derivation of MSCs and their differentiation capability into specific cell lineages at the transplanted site. Therefore, this review is focussed on precise mechanisms by which transplantation of MSCs promote functional recovery and also addresses the challenges to improve the therapeutic efficacy of MSCs for treating SCI in dogs.

Heat-Shock Proteins Can Potentiate the Therapeutic Ability of Cryopreserved Mesenchymal Stem Cells for the Treatment of Acute Spinal Cord Injury in Dogs.

Mesenchymal stem cells (MSCs) are applied in the treatment of spinal cord injury (SCI) because of their neural tissue restoring ability. In the clinical setting, intravenous injection of cryopreserved cells is essential for the immediate treatment of SCI, exhibiting the disadvantage of reduced cell properties. In this study, we potentiated the characteristics of cryopreserved MSCs by heat-shock (HS) treatment to induce the expression of HS protein (HSP) HSP70/HSP27 and further improved antioxidant capacity by overexpressing HSP32 (heme oxygenase-1 [HO-1]). We randomly assigned 12 beagle dogs with acute SCI into three groups and transplanted cells intravenously: (i) F-MSCs (MSCs in frozen/thawed conditions); (ii) F-HSP-MSCs (HS-treated MSCs in frozen/thawed conditions); and (iii) F-HSP-HO-MSCs (HO-1-overexpressing and HS-treated MSCs in frozen/thawed conditions). The potentiated MSCs exhibited increased growth factor-, anti-inflammatory-, antioxidant-, homing- and stemness-related gene expression. In the animal experiments, the HSP-induced groups showed significant improvement in hind-limb locomotion, highly expressed neural markers, less intervened fibrotic changes, and improved myelination. In particular, the HO-1-overexpression group was more prominent, controlling the initial inflammatory response with high antioxidant capabilities, suggesting that antioxidation was important to prevent secondary injury. Accordingly, HSPs not only successfully increased the ability of frozen MSCs but also demonstrated excellent neural protection and regeneration capacity in the case of acute SCI. The application of HSP-induced cryopreserved MSCs in first-aid treatment for acute SCI is considered to help early neural sparing and further hind-limb motor function restoration.

Treatment of Chronic Spinal Cord Injury in Dogs Using Amniotic Membrane-Derived Stem Cells: Preliminary Results.

IntroductionIntervertebral disc diseases (IVDD) represent the majority of neurological attendance and responsible for the most cases of paralysis in dogs. Treatments currently used do not show satisfactory results in patients with more severe and chronic neurological manifestations.MethodsTo promote nerve and muscular recovery, as well as improve quality of life, we aimed to create a double-blind test method, associating spinal decompression surgery and allogeneic transplantation of amniotic membrane-derived stem cells (AMSCs) in dogs with chronic IVDD. Cells were characterized as fetal mesenchymal cells and safe for application. Eight animals completed the experiment: stem cell applications were made in four animals that had previously undergone an unsuccessful surgical procedure (“SC group”, n = 4); two animals were submitted to surgery, followed by applications of stem cells (“Surgery + SC”, n = 2); two other animals were submitted to surgery, followed by the application of saline solution (“Surgery + placebo”, n = 2). During the surgical procedure, a topical application was performed on the lesion and after fifteen and forty-five days another two applications were made via epidural. Animals were monitored biweekly and reassessed three months after surgery, by functional tests and magnetic resonance exams.ResultsSome animals presented significant neurological improvement, such as the recovery of nociception and ability to remain on station. Despite the need further studies, until the present moment, cell therapy has been feasible and has no harmful effects on animals.ConclusionThe protocol of preclinical trial showed the association with decompressive surgery and cell transplantation in dogs with thoracolumbar IVDD proved feasible, and it was possible to observe neurological improvement after treatment. No tissue improvement through MRI was found. The double-blind test guaranteed reliability of the evaluations and results obtained that, even with a small sample size, generated satisfactory results for the animals and owners.

Current Insights Into the Pathology of Canine Intervertebral Disc Extrusion-Induced Spinal Cord Injury.

Spinal cord injury (SCI) in dogs is commonly attributed to intervertebral disc extrusion (IVDE). Over the last years substantial progress was made in the elucidation of factors contributing to the pathogenesis of this common canine disease. A detailed understanding of the underlying histopathological and molecular alterations in the lesioned spinal cord represents a prerequisite to translate knowledge on the time course of secondary injury processes into the clinical setting. This review summarizes the current state of knowledge of the underlying pathology of canine IVDE-related SCI. Pathological alterations in the spinal cord of dogs affected by IVDE-related SCI include early and persisting axonal damage and glial responses, dominated by phagocytic microglia/macrophages. These processes are paralleled by a pro-inflammatory microenvironment with dysregulation of cytokines and matrix metalloproteinases within the spinal cord. These data mirror findings from a clinical and therapeutic perspective and can be used to identify biomarkers that are able to more precisely predict the clinical outcome. The pathogenesis of progressive myelomalacia, a devastating complication of SCI in dogs, is not understood in detail so far; however, a fulminant and exaggerating secondary injury response with massive reactive oxygen species formation seems to be involved in this unique neuropathological entity. There are substantial gaps in the knowledge of pathological changes in IVDE with respect to more advanced and chronic lesions and the potential involvement of demyelination. Moreover, the role of microglia/macrophage polarization in IVDE-related SCI still remains to be investigated. A close collaboration of clinical neurologists and veterinary pathologists will help to facilitate an integrative approach to a more detailed understanding of the molecular pathogenesis of canine IVDE and thus to identify therapeutic targets.

Ambulation in Dogs With Absent Pain Perception After Acute Thoracolumbar Spinal Cord Injury

Acute thoracolumbar spinal cord injury (SCI) is common in dogs frequently secondary to intervertebral disc herniation. Following severe injury, some dogs never regain sensory function to the pelvic limbs or tail and are designated chronically “deep pain negative.” Despite this, a subset of these dogs develop spontaneous motor recovery over time including some that recover sufficient function in their pelvic limbs to walk independently without assistance or weight support. This type of ambulation is commonly known as “spinal walking” and can take up to a year or more to develop. This review provides a comparative overview of locomotion and explores the physiology of locomotor recovery after severe SCI in dogs. We discuss the mechanisms by which post-injury plasticity and coordination between circuitry contained within the spinal cord, peripheral sensory feedback, and residual or recovered supraspinal connections might combine to underpin spinal walking. The clinical characteristics of spinal walking are outlined including what is known about the role of patient or injury features such as lesion location, timeframe post-injury, body size, and spasticity. The relationship between the emergence of spinal walking and electrodiagnostic and magnetic resonance imaging findings are also discussed. Finally, we review possible ways to predict or facilitate recovery of walking in chronically deep pain negative dogs. Improved understanding of the mechanisms of gait generation and plasticity of the surviving tissue after injury might pave the way for further treatment options and enhanced outcomes in severely injured dogs.

Recombinant canine basic fibroblast growth factor-induced differentiation of canine bone marrow mesenchymal stem cells into voltage- and glutamate-responsive neuron-like cells.

IntroductionBasic fibroblast growth factor (bFGF) is a promising cytokine in regenerative therapy for spinal cord injury. In this study, recombinant canine bFGF (rc-bFGF) was synthesized for clinical use in dogs, and the ability of rc-bFGF to differentiate canine bone marrow mesenchymal stem cells (BMSCs) into functional neurons was investigated.MethodsThe rc-bFGF was synthesized using a wheat germ cell-free protein synthesis system. The expression of rc-bFGF mRNA in the purification process was confirmed using a reverse transcription-polymerase chain reaction (RT-PCR). Western blotting was performed to confirm the antigenic property of the purified protein. To verify function of the purified protein, phosphorylation of extracellular signal-regulated kinase (ERK) was examined by in vitro assay using HEK293 cells. To compare the neuronal differentiation capacity of canine BMSCs in response to treatment with rc-bFGF, the cells were divided into the following four groups: control, undifferentiated, rh-bFGF, and rc-bFGF groups. After neuronal induction, the percentage of cells that had changed to a neuron-like morphology and the mRNA expression of neuronal markers were evaluated. Furthermore, to assess the function of the canine BMSCs after neuronal induction, changes in the intracellular Ca2+ concentrations after stimulation with KCl and l-glutamate were examined.ResultsThe protein synthesized in this study was rc-bFGF and functioned as bFGF, from the results of RT-PCR, western blotting, and the expression of pERK in HEK293 cells. Canine BMSCs acquired a neuron-like morphology and expressed mRNAs of neuronal markers after neuronal induction in the rh-bFGF and the rc-bFGF groups. These results were more marked in the rc-bFGF group than in the other groups. Furthermore, an increase in intracellular Ca2+ concentrations was observed after the stimulation of KCl and l-glutamate in the rc-bFGF group, same as in the rh-bFGF group.ConclusionsA functional rc-bFGF was successfully synthesized, and rc-bFGF induced the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells. Our purified rc-bFGF may contribute, on its own, or in combination with canine BMSCs, to regenerative therapy for spinal cord injury in dogs.

Long-term postoperative pain evaluation in dogs with thoracolumbar intervertebral disk herniation after hemilaminectomy.

BackgroundChronic neuropathic pain is a common complication in people with spinal cord injury (SCI) but has not been investigated in dogs.ObjectiveTo determine the reliability of measuring spinal mechanical sensory thresholds (MSTs) in dogs and to compare MSTs of healthy dogs and dogs with SCI caused by acute thoracolumbar intervertebral disk extrusion after hemilaminectomy over a 1‐year period.Study DesignProspective study.AnimalsThirty‐two healthy and 40 SCI dogs.MethodsDogs were divided into group 1 (healthy Dachshunds), group 2 (healthy dogs including several breeds), and SCI group. The MSTs were measured using algometry at an incision (thoracolumbar) and control site. Dogs in group 1 were tested once; those in group 2 were tested for 5 consecutive days; and SCI dogs were tested on days 7, 14, 28, 42, 180, and 365 postoperatively. The MSTs were compared among days in healthy and SCI dogs and between SCI and healthy dogs using mixed effect models. P < .05 was considered significant.ResultsAt the incision site of SCI dogs, MST was significantly lower than in healthy dogs for 42 days postoperatively, but not subsequently. However, 4/27 dogs had control site MST below the reference range 1 year after surgery.Conclusions and Clinical ImportanceMechanical sensory thresholds normalize by 6 months after surgery in most dogs with SCI. Approximately 15% of SCI dogs may develop chronic neuropathic pain. Improving long‐term pain assessment of SCI dogs is important for offering treatment options and advising owners.

Retrospective evaluation of prazosin and diazepam after thoracolumbar hemilaminectomy in dogs

Management of urinary retention after spinal cord injury in dogs is often needed and can include use of medications to relax the urethral sphincter. This was a retrospective study evaluating two such medications, prazosin and diazepam, and whether dogs treated with these medications had different lengths of hospitalization, urinary continence levels, or development of bacteriuria compared to dogs not receiving these medications after thoracolumbar hemilaminectomy for intervertebral disc herniation (IVDH). Electronic medical records were searched for dogs that underwent CT or MRI followed by a hemilaminectomy between the 3rd thoracic and 3rd lumbar vertebra for treatment of IVDH. Dogs were grouped based on whether or not they received a medication to aid in urethral sphincter relaxation (either prazosin, diazepam, or both medications). The total length of hospitalization, urinary continence at the time of discharge, and presence of bacteriuria were recorded from the medical file. Medical records from 71 dogs were included in the analysis.There were no significant associations between administration of prazosin and/or diazepam and length of hospitalization or urinary continence scores at the time of discharge from the hospital (P > 0.05).

Development of a Novel Gait Analysis Tool Measuring Center of Pressure for Evaluation of Canine Chronic Thoracolumbar Spinal Cord Injury

Gait evaluation after spinal cord injury (SCI) is an important component of determining functional status. Analysis of center of pressure (COP) provides a dynamic reflection of global locomotion and postural control and has been used to quantify various gait abnormalities. We hypothesized that COP variability would be greater for SCI versus normal dogs and that COP would be able to differentiate varying injury severity. Our objective was to investigate COP, COP variability, and body weight support percentage in dogs with chronic SCI. Eleven chronically non-ambulatory dogs after acute severe thoracolumbar SCI were enrolled. COP measurements in x (right-to-left, COPx) and y (craniocaudal, COPy) directions were captured while dogs walked on a pressure-sensitive treadmill with pelvic limb sling support. Root mean square values (RMS_COPx and RMS_COPy) were calculated to assess variability in COP. Body weight support percentage was measured using a load cell. Gait also was quantified using an open field scale (OFS) and treadmill-based stepping and coordination scores (SS, RI). Mean COPx, COPy, RMS_COPx, and RMS_COPy were compared between dogs with SCI and previously evaluated healthy controls. RMS measurements and support percentage were compared with standard gait scales (OFS, SS, RI). Mean COPy was more cranial and RMS_COPx and RMS_COPy were greater in SCI versus normal dogs (p < 0.001). Support percentage moderately correlated with SS (p = 0.019; R2 = 0.47). COP analysis and body weight support measurements offer information about post-injury locomotion. Further development is needed before consideration as an outcome measure to complement validated gait analysis methods in dogs with SCI.

Improved Healing after the Co-Transplantation of HO-1 and BDNF Overexpressed Mesenchymal Stem Cells in the Subacute Spinal Cord Injury of Dogs

Abundant expression of proinflammatory cytokines after a spinal cord injury (SCI) creates an inhibitory microenvironment for neuroregeneration. The mesenchymal stem cells help to mitigate the inflammation and improve neural growth and survival. For this purpose, we potentiated the function of adipose-derived mesenchymal stem cells (Ad-MSCs) by transfecting them with brain-derived neurotrophic factor (BDNF) and heme oxygenase-1 (HO-1), through a lentivirus, to produce BDNF overexpressed Ad-MSCs (BDNF-MSCs), and HO-1 overexpressed Ad-MSCs (HO-1-MSCs). Sixteen SCI beagle dogs were randomly assigned into four treatment groups. We injected both HO-1 and BDNF-overexpressed MSCs as a combination group, to selectively control inflammation and induce neuroregeneration in SCI dogs, and compared this with BDNF-MSCs, HO-1-MSCs, and GFP-MSCs injected dogs. The groups were compared in terms of improvement in canine Basso, Beattie, and Bresnahan (cBBB) score during 8 weeks of experimentation. After 8 weeks, spinal cords were harvested and subjected to western blot analysis, immunofluorescent staining, and hematoxylin and eosin (H&E) staining. The combination group showed a significant improvement in hindlimb functions, with a higher BBB score, and a robust increase in neuroregeneration, depicted by a higher expression of Tuj-1, NF-M, and GAP-43 due to a decreased expression of the inflammatory markers interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), and an increased expression of interleukin-10 (IL-10) (P ≤ 0.05). H&E staining showed more reduced intraparenchymal fibrosis in the combination group than in other groups (P ≤ 0.05). It was thus suggested that the cotransplantation of HO-1 and BDNF-MSCs is more effective in promoting the healing of SCI. HO-1-MSCs reduce inflammation, which favors BDNF-induced neuroregeneration in SCI of dogs.

Diffusion Tensor Imaging of Scarring, Necrosis, and Cavitation Based on Histopathological Findings in Dogs with Chronic Spinal Cord Injury: Evaluation of Multiple Diffusion Parameters and Their Correlations with Histopathological Findings.

The use of diffusion tensor imaging (DTI) for the characterization of various lesion types in dogs with spinal cord injury (SCI) has not been investigated. The aim of this study was to characterize scarring (loose immature scarring [LIMS], intermediate mature scarring [IMS], and dense mature scarring [DMS]), necrosis, cavitation, and acute hemorrhage using multiple DTI parameters and determine the correlations between the DTI parameters and histopathological finding in dogs with controlled SCI. All imaging data were obtained from the lumbar spinal cord (from L1 to L3) of normal and SCI dogs using a 3-Tesla magnetic resonance imaging scanner. Transverse multi-shot echo planar imaging (EPI) sequences (12 directions; b-value, 0 and 800 s/mm2) were used for DTI. Regions of interest on DTI maps were selected according to the areas of normal white matter (NWM), normal gray matter (NGM), LIMS, IMS, DMS, necrosis, and cavitation on histopathological images and the area of acute hemorrhage with a hypointense signal on T2*-weighted images obtained 24 h post-SCI. There were statistically significant differences in DTI parameters among NWM, NGM, multi-grade scarring, necrosis, and cavitation and between mature scarring and acute hemorrhage. The maturation grade of scarring demonstrated a positive linear correlation with fractional anisotropy and the planar index and a negative linear correlation with the spherical index and the radial, mean, and axial diffusivities. These results suggest the feasibility of using DTI for detailed noninvasive monitoring of SCI. DTI can provide critical information for guiding therapeutic strategies and determining the prognosis of SCI patients.

Transplantation of canine olfactory ensheathing cells producing chondroitinase ABC promotes chondroitin sulphate proteoglycan digestion and axonal sprouting following spinal cord injury.

Olfactory ensheathing cell (OEC) transplantation is a promising strategy for treating spinal cord injury (SCI), as has been demonstrated in experimental SCI models and naturally occurring SCI in dogs. However, the presence of chondroitin sulphate proteoglycans within the extracellular matrix of the glial scar can inhibit efficient axonal repair and limit the therapeutic potential of OECs. Here we have used lentiviral vectors to genetically modify canine OECs to continuously deliver mammalian chondroitinase ABC at the lesion site in order to degrade the inhibitory chondroitin sulphate proteoglycans in a rodent model of spinal cord injury. We demonstrate that these chondroitinase producing canine OECs survived at 4 weeks following transplantation into the spinal cord lesion and effectively digested chondroitin sulphate proteoglycans at the site of injury. There was evidence of sprouting within the corticospinal tract rostral to the lesion and an increase in the number of corticospinal axons caudal to the lesion, suggestive of axonal regeneration. Our results indicate that delivery of the chondroitinase enzyme can be achieved with the genetically modified OECs to increase axon growth following SCI. The combination of these two promising approaches is a potential strategy for promoting neural regeneration following SCI in veterinary practice and human patients.

Prognostic Factors Associated with Recovery of Ambulation and Urinary Continence in Dogs with Acute Lumbosacral Spinal Cord Injury.

BackgroundLimited information is available about prognostic factors for recovery after spinal cord injury (SCI) to the L4‐S3 segments. Previous research suggests that L4‐S3 SCI does not have a worse prognosis than T3‐L3 SCI.Hypothesis/ObjectivesTo elucidate prognostic factors for regaining urinary continence and ambulation in dogs with L4‐S3 SCI and compare prognosis to T3‐L3 SCI.Animals/MethodsA retrospective study on 61 nonambulatory dogs with L4‐S3 SCI, matched to dogs with T3‐L3 SCI, compared 3 weeks after onset. Prognostic factors explored using logistic regression and used for matching: nonchondrodystrophic dogs >15 kg versus dogs that were chondrodystrophic or <15 kg; compressive versus noncompressive lesions; presence versus absence of conscious pain perception (CPP); and lower vs upper motor neuron (LMN/UMN) incontinence.ResultsFewer L4‐S3 dogs regained continence compared to T3‐L3 dogs (64 vs 85%, P = .0033), but no difference existed for regaining ambulation (66 vs 75%, P = .1306). In L4‐S3 SCI dogs, fewer dogs regained continence with loss of CPP (P < .001), LMN incontinence (P = .004), and noncompressive lesions (P = .006). Negative prognostic factors for regaining ambulation included absent CPP (P < .001) and large nonchondrodystrophic breed (P = .022).Conclusions and Clinical ImportanceDogs with L4‐S3 SCI have a poorer short‐term prognosis than do dogs with T3‐L3 SCI. Dogs with L4‐S3 SCI had a poor prognosis with loss of CPP, or noncompressive lesions combined with LMN incontinence. Small‐breed or chondrodystrophic dogs with retained CPP, compressive lesions, and UMN incontinence had an excellent prognosis. These findings may help guide decision‐making in L4‐S3 SCI.

Arachidonic acid pathway alterations in cerebrospinal fluid of dogs with naturally occurring spinal cord injury.

BackgroundCanine intervertebral disc πherniation causes a naturally-occurring spinal cord injury (SCI) that bears critical similarities to human SCI with respect to both injury pathomechanisms and treatment. As such, it has tremendous potential to enhance our understanding of injury biology and the preclinical evaluation of novel therapies. Currently, there is limited understanding of the role of arachidonic acid metabolites in canine SCI.ResultsThe CSF concentrations of PLA2 and PGE2 were higher in SCI dogs compared to control dogs (p = 0.0370 and 0.0273, respectively), but CSF LCT4 concentration in SCI dogs was significantly lower than that in control dogs (p < 0.0001). Prostaglandin E2 concentration in the CSF was significantly and positively associated with increased severity of SCI at the time of sampling (p = 0.041) and recovery 42 days post-injury (p = 0.006), as measured by ordinal behavioral scores.ConclusionArachidonic acid metabolism is altered in dogs with SCI, and these data suggest that these AA metabolites reflect injury severity and recovery, paralleling data from other model systems.

Adaptation of the Basso–Beattie–Bresnahan locomotor rating scale for use in a clinical model of spinal cord injury in dogs

BackgroundNaturally occurring acute spinal cord injury (SCI) in pet dogs provides an important clinical animal model through which to confirm and extend findings from rodent studies; however, validated quantitative outcome measures for dogs are limited. New methodWe adapted the Basso Beattie Bresnahan (BBB) scale for use in a clinical dog model of acute thoracolumbar SCI. Based on observation of normal dogs, modifications were made to account for species differences in locomotion. Assessments of paw and tail position, and trunk stability were modified to produce a 19 point scale suitable for use in dogs, termed the canine BBB scale (cBBB). Pet dogs with naturally occurring acute SCI were assigned cBBB scores at 3, 10 and 30days after laminectomy. ResultsScores assigned via the cBBB were stable across testing sessions in normal dogs but increased significantly between days 3 and 30 in SCI-affected dogs (p=0.0003). The scale was highly responsive to changes in locomotor recovery over a 30day period, with a standardized response mean of 1.34. Comparison with existing methodsConcurrent validity was good, with strong correlations observed between the cBBB and two other locomotor scales, the OSCIS (r=0.94; p<0.001) and the MFS (r=0.85; p<0.0001). cBBB scores inversely correlated with other assessments of recovery including mechanical sensory threshold (r=−0.68; p<0.0001) and coefficient of variation of stride length (r=−0.49; p<0.0001). ConclusionsThese results support the use of the cBBB to assess locomotor recovery in canine clinical translational models of SCI.