Hendrik-Jan C. Kranenburg

The Dog as an Animal Model for Intervertebral Disc Degeneration

Study Design. Prospective observational and analytic study. Objective. To investigate whether spontaneous intervertebral disc degeneration (IVDD) occurring in both chondrodystrophic (CD) and nonchondrodystrophic dogs (NCD) can be used as a valid translational model for human IVDD research. Summary of Background Data. Different animal models are used in IVDD research, but in most of these models IVDD is induced manually or chemically rather than occurring spontaneously. Methods. A total of 184 intervertebral discs (IVDs) from 19 dogs of different breeds were used. The extent of IVDD was evaluated by macroscopic grading, histopathology, glycosaminoglycan content, and matrix metalloproteinase 2 activity. Canine data were compared with human IVD data acquired in this study or from the literature. Results. Gross pathology of IVDD in both dog types (CD and NCD) and humans showed many similarities, but the cartilaginous endplates were significantly thicker and the subchondral cortices significantly thinner in humans than in dogs. Notochordal cells were still present in the IVDs of adult NCD but were not seen in the CD breeds or in humans. Signs of degeneration were seen in young dogs of CD breeds (<1 year of age), whereas this was only seen in older dogs of NCD breeds (5–7 years of age). The relative glycosaminoglycan content and metalloproteinase 2 activity in canine IVDD were similar to those in humans: metalloproteinase 2 activity increased and glycosaminoglycan content decreased with increasing severity of IVDD. Conclusion. IVDD is similar in humans and dogs. Both CD and NCD breeds may therefore serve as models of spontaneous IVDD for human research. However, as with all animal models, it is important to recognize interspecies differences and, indeed, the intraspecies differences between CD and NCD breeds (early vs. late onset of IVDD, respectively) to develop an optimal canine model of human IVDD.

Intervertebral disc disease in dogs – Part 2: Comparison of clinical, magnetic resonance imaging, and histological findings in 74 surgically treated dogs

The relationship between intervertebral disc (IVD) disease and IVD degeneration remains unclear. The aim of the present study was to compare the clinical severity of IVD herniation (IVDH), determined with a neurological grading system, with findings of magnetic resonance imaging (MRI) and histology using grading systems for IVD degeneration in chondrodystrophic (CD; n=37) and non-chondrodystrophic (NCD; n=37) dogs. This study is the second part of a two-part investigation, where the first part involved the development and validation of a histological grading scheme for classification of canine IVD degeneration. IVD degeneration graded on MRI correlated significantly with IVD degeneration graded on histology, but not with pre-operative clinical signs. Hansen type 1 hernias were more common in the cervical and thoracolumbar segments and Hansen type 2 hernias were more common in the lumbosacral segment. Type 1 hernias occurred more often in CD dogs than in NCD dogs, and CD dogs were clinically more severely affected than NCD dogs. The grade of IVD degeneration on MRI was higher in CD dogs than in NCD dogs, but there was no difference between dogs with type 1 and type 2 hernias. No significant differences in histological grade were found between CD and NCD dogs or between type 1 and type 2 hernias. It was possible to conclude that IVD degeneration did not correlate with the neurological severity of IVDH. The extent of degeneration identified on MRI correlated with degeneration seen histologically. Although the MRI grading system reflected the severity of IVD degenerative changes as confirmed by histopathology, it appeared less useful in predicting the clinical implications.

An annulus fibrosus closure device based on a biodegradable shape-memory polymer network

Injuries to the intervertebral disc caused by degeneration or trauma often lead to tearing of the annulus fibrosus (AF) and extrusion of the nucleus pulposus (NP). This can compress nerves and cause lower back pain. In this study, the characteristics of poly(D,L-lactide-co-trimethylene carbonate) networks with shape-memory properties have been evaluated in order to prepare biodegradable AF closure devices that can be implanted minimally invasively. Four different macromers with (D,L-lactide) to trimethylene carbonate (DLLA:TMC) molar ratios of 80:20, 70:30, 60:40 and 40:60 with terminal methacrylate groups and molecular weights of approximately 30 kg mol(-1) were used to prepare the networks by photo-crosslinking. The mechanical properties of the samples and their shape-memory properties were determined at temperatures of 0 °C and 40 °C by tensile tests- and cyclic, thermo-mechanical measurements. At 40 °C all networks showed rubber-like behavior and were flexible with elastic modulus values of 1.7-2.5 MPa, which is in the range of the modulus values of human annulus fibrosus tissue. The shape-memory characteristics of the networks were excellent with values of the shape-fixity and the shape-recovery ratio higher than 98 and 95%, respectively. The switching temperatures were between 10 and 39 °C. In vitro culture and qualitative immunocytochemistry of human annulus fibrosus cells on shape-memory films with DLLA:TMC molar ratios of 60:40 showed very good ability of the networks to support the adhesion and growth of human AF cells. When the polymer network films were coated by adsorption of fibronectin, cell attachment, cell spreading, and extracellular matrix production was further improved. Annulus fibrosus closure devices were prepared from these AF cell-compatible materials by photo-polymerizing the reactive precursors in a mold. Insertion of the multifunctional implant in the disc of a cadaveric canine spine showed that these shape-memory devices could be implanted through a small slit and to some extent deploy self-sufficiently within the disc cavity.

The dog as an animal model for DISH

Diffuse idiopathic skeletal hyperostosis (DISH) is a systemic disorder of the axial and peripheral skeleton in humans and has incidentally been described in dogs. The aims of this retrospective radiographic cohort study were to determine the prevalence of DISH in an outpatient population of skeletally mature dogs and to investigate if dogs can be used as an animal model for DISH. The overall prevalence of canine DISH was 3.8% (78/2041). The prevalence of DISH increased with age and was more frequent in male dogs, similar to findings in human studies. In the Boxer breed the prevalence of DISH was 40.6% (28/69). Dog breeds represent closed gene pools with a high degree of familiar relationship and the high prevalence in the Boxer may be indicative of a genetic origin of DISH. It is concluded that the Boxer breed may serve as an animal model for DISH in humans.

Diffuse idiopathic skeletal hyperostosis (DISH) and spondylosis deformans in purebred dogs: a retrospective radiographic study.

A retrospective radiographic study was performed to investigate the prevalence of diffuse idiopathic skeletal hyperostosis (DISH) and spondylosis deformans (spondylosis) in 2041 purebred dogs and to determine association with age, gender and breed. Four cases of DISH provided information on the appearance of canine DISH. The prevalence of DISH and spondylosis was 3.8% (78/2041) and 18.0% (367/2041), respectively. Of dogs with DISH, 67.9% (53/78) also had spondylosis, whereas 14.0% (53/367) of dogs with spondylosis also had DISH. Dogs with DISH and/or spondylosis were significantly older than those without spinal exostosis. The prevalence of DISH and spondylosis was 40.6% (28/69) and 55.1% (38/69), respectively, in Boxer dogs. Nineteen smaller breeds were not affected by DISH, but showed signs of spondylosis; only standard Poodles appeared not to be affected by either disorder. Radiography, computed tomography (CT), magnetic resonance imaging (MRI), and/or histopathology were used to investigate four DISH cases. It was concluded that spondylosis and DISH can co-occur in dogs. DISH has probably been previously under-diagnosed and mistaken for severe spondylosis. The diagnosis can be made using radiography, CT or MRI. On histology, DISH can be distinguished from spondylosis by the location (ventral longitudinal ligament) and extent of new bone formation.

Biocompatibility and intradiscal application of a thermoreversible celecoxib-loaded poly-N-isopropylacrylamide MgFe-layered double hydroxide hydrogel in a canine model.

IntroductionChronic low back pain due to intervertebral disc (IVD) degeneration is associated with increased levels of inflammatory mediators. Current medical treatment consists of oral anti-inflammatory drugs to alleviate pain. In this study, the efficacy and safety of a novel thermoreversible poly-N-isopropylacrylamide MgFe-layered double hydroxide (pNIPAAM MgFe-LDH) hydrogel was evaluated for intradiscal controlled delivery of the selective cyclooxygenase (COX) 2 inhibitor and anti-inflammatory drug celecoxib (CXB).MethodsDegradation, release behavior, and the ability of a CXB-loaded pNIPAAM MgFe-LDH hydrogel to suppress prostaglandin E2 (PGE2) levels in a controlled manner in the presence of a proinflammatory stimulus (TNF-α) were evaluated in vitro. Biocompatibility was evaluated histologically after subcutaneous injection in mice. Safety of intradiscal application of the loaded and unloaded hydrogels was studied in a canine model of spontaneous mild IVD degeneration by histological, biomolecular, and biochemical evaluation. After the hydrogel was shown to be biocompatible and safe, an in vivo dose–response study was performed in order to determine safety and efficacy of the pNIPAAM MgFe-LDH hydrogel for intradiscal controlled delivery of CXB.ResultsCXB release correlated to hydrogel degradation in vitro. Furthermore, controlled release from CXB-loaded hydrogels was demonstrated to suppress PGE2 levels in the presence of TNF-α. The hydrogel was shown to exhibit a good biocompatibility upon subcutaneous injection in mice. Upon intradiscal injection in a canine model, the hydrogel exhibited excellent biocompatibility based on histological evaluation of the treated IVDs. Gene expression and biochemical analyses supported the finding that no substantial negative effects of the hydrogel were observed. Safety of application was further confirmed by the absence of clinical symptoms, IVD herniation or progression of degeneration. Controlled release of CXB resulted in a nonsignificant maximal inhibition (approximately 35 %) of PGE2 levels in the mildly degenerated canine IVDs.ConclusionsIn conclusion, this study showed biocompatibility and safe intradiscal application of an MgFe LDH-pNIPAAM hydrogel. Controlled release of CXB resulted in only limited inhibition of PGE2 in this model with mild IVD degeneration, and further studies should concentrate on application of controlled release from this type of hydrogel in animal models with more severe IVD degeneration.

Prevalence of spondylosis deformans in the feline spine and correlation with owner-perceived behavioural changes

OBJECTIVES The primary objective was to determine the prevalence, spinal distribution, and association with the signalment of cats suffering from different grades of feline spondylosis deformans (spondylosis). The secondary objective was to document behavioural changes associated with spondylosis by owner observation. METHODS A cross-sectional study was performed to determine the prevalence of feline spondylosis (group 1). A prospective study was performed to determine the association between radiographic abnormalities of the lumbosacral region (L3-S1) and owner perceived behavioural changes based on a completed questionnaire (group 2). The radiographs were reviewed using a grading system (0-3) for spondylosis. RESULTS The prevalence of spondylosis in group 1 was 39.4% (158/402). Cats with spondylosis were significantly older than cats without spondylosis (p <0.001). The thoracic (T) vertebrae T4-T10 were most often affected by spondylosis, but spondylosis was most severe in the T10-S1 vertebrae. In group 2, spondylosis of the lumbosacral region was significantly correlated with owner-reported behavioural changes, such as a decreased willingness to greet people and to being petted, increased aggressiveness, and a poor perceived quality of life (p = 0.037). CLINICAL SIGNIFICANCE This study found that feline spondylosis is common and that spondylosis of the lumbosacral region may be accompanied by behavioural changes.

Spinal hyperostosis in humans and companion animals

Both spondylosis and diffuse idiopathic skeletal hyperostosis (DISH) are prevalent in humans and are considered distinct entities. Nowadays, the term spondylosis is in the biomedical literature mostly used when also degenerative disc disease is present. In companion animals, many reports on spondylosis, often without intervertebral disc degeneration, are described. The nomenclature and the definitions of both spondylosis and DISH in the biomedical and veterinary literature should be more in line to facilitate comparison. DISH occurs in dogs but has not been described in cats yet. DISH and spondylosis can co-occur in dogs in one animal. Boxers may serve as translational disease models for the elucidation of the gene(s) involved in the (etio)pathogenesis of DISH or serve as a test population for newly developed treatment options.

Design, synthesis, imaging, and biomechanics of a softness-gradient hydrogel nucleus pulposus prosthesis in a canine lumbar spine model

A hydrogel nucleus pulposus prosthesis (NPP) was designed to swell in situ, have intrinsic radiopacity, and restore intervertebral disc height and biomechanical functionality. These features were examined using an ex vivo canine lumbar model. Nine NPPs were implanted in five spines and their visibility was assessed on radiography, computed tomography (CT), and magnetic resonance imaging (MRI). The NPPs were visible on all imaging modalities and 8/9 NPPs stayed intact and in situ. Six other NPPs were tested biomechanically in six canine lumbar spines. Removal of the nucleus pulposus (nuclectomy) caused significant changes in biomechanical parameters. After implantation and swelling of the NPP, values were not significantly different from the native state for range of motion (ROM) of flexion-extension (FE) and lateral bending (LB), the neutral zone (NZ) of all motion directions, and the NZ stiffness (NZS) of FE. Biomechanical restoration by the NPP compared with the nuclectomized state was significant for the ROM of FE and axial rotation, the NZ of FE and LB, and the NZS of FE and LB. Disc height was significantly restored and 6/6 NPPs stayed intact and in situ. In conclusion, the NPPs swell in situ, have intrinsic radiopacity and restored disc height and aforementioned biomechanical properties.

Naturally Occurring Spinal Hyperostosis in Dogs as a Model for Human Spinal Disorders

Both spondylosis and diffuse idiopathic skeletal hyperostosis (DISH) are prevalent in humans and are considered distinct entities. Nowadays, the term spondylosis is in the biomedical literature mostly used when concurrently degenerative disc disease is present. In companion animals, many reports on spondylosis, often without intervertebral disc degeneration, are described. The nomenclature and the definitions of both spondylosis and DISH in biomedical and veterinary literature should be more in line to facilitate comparison. Spondylosis and DISH occur in dogs spontaneously and can co-occur in one animal. Specifically, Boxers may serve as translational disease models for the elucidation of the gene(s) involved in the (etio)pathogenesis of spondylosis and DISH or serve as a test population for newly developed treatment options.