Genome-wide association study identifies genomic regions of association for cruciate ligament rupture in Newfoundland dogs.

Cranial cruciate ligament rupture (CCLR) is the most common cause of pelvic limb lameness in dogs. To investigate the genetic basis of canine CCLR, we conducted a genome-wide association study using a canine SNP array in Newfoundland pedigree dogs with and without CCLR (n = 96). We identified three main chromosomal regions of CCLR association (on chromosomes 1, 3 and 33). Each of these regions was confirmed by Sequenom genotyping in a further cohort of Newfoundlands (n = 271). The results, particularly SNPs identified in the SORCS2 and SEMA5B genes, suggest that there may be neurological pathways involved in susceptibility to canine CCLR.

Prevalence of radiographic appendicular osteoarthritis and associated clinical signs in young dogs.

This study aimed to determine the prevalence of osteoarthritis (OA) and associated clinical signs in young dogs. Owners of dogs aged 8 months-4 years from a single practice, were contacted in random order, to participate in a general health screen. Clinical and orthopedic examinations were performed. Each joint was scored for pain reactions (0-4). Orthogonal radiographs of all joints were made under sedation. Each joint was scored for radiographic OA (rOA) severity on an 11-point scale. Clinical OA (cOA) was defined as an overlap of rOA and joint pain in ≥ 1 joint. Owners completed OA questionnaires. The owners of 123 dogs agreed to participate. Overall, 39.8% (49/123) of dogs had rOA in ≥ 1 joint, and 16.3% (20/123) or 23.6% (29/123) dogs had cOA, depending on the cut-off value of joint pain; moderate (2), or mild (1), respectively. Owners of dogs with cOA observed signs of impairment in approximately 30% of cases. Only 2 dogs with cOA were receiving OA pain management. The most commonly affected joints in descending order of frequency were elbow, hip, tarsus, and stifle. Radiographically visible OA is common in young dogs, and 40-60% of dogs with rOA had cOA. However, OA-pain appears underdiagnosed and undertreated in young dogs.

Observer variation in the evaluation and classification of severe central tarsal bone fractures in racing Greyhounds.

OBJECTIVES: To determine observer agreement on radiographic evaluation of central tarsal bone (CTB) fractures and compare this with evaluation of the same fractures using computed tomography (CT). METHODS: Radiographs and CT scans were obtained of the right tarsi from limbs of Greyhounds euthanatized after sustaining severe CTB fracture during racing. Four observers described and classified each fracture. Inter- and intra-observer agreements were calculated. RESULTS: Inter-observer agreement was higher for assessment of fractures using CT. Several fractures assessed by radiography were misclassified as a less severe type. Intra-observer agreement for assessment and classification of CTB fractures via radiography versus CT was variable. Overall agreement among all four observers was higher for CT than radiography. Additionally, when identifying fractures of the adjacent tarsal bones, observer agreement was higher for CT than radiography. CLINICAL SIGNIFICANCE: Computed tomography improved observer ability to correctly evaluate CTB fracture and detect the degree of displacement and extent of any comminution. Identification of fractures of adjacent tarsal bones was also improved when tarsi were assessed using CT. These data suggest that treatment decisions based solely on radiographic assessment of CTB fractures may not produce the expected outcome.

Demineralised bone matrix in veterinary orthopaedics: a review.

Demineralised bone matrix (DBM) is commonly used in human orthopaedics as an allograft prepared from cortical bone. As such, there is a background of literature on the basic science, experimental animal studies and clinical human use of DBM. Because canine DBM is now increasingly available and used in veterinary orthopaedics, this review aims to update the veterinary orthopaedic specialist with the properties and activities of this bone allograft product.

A humeral intracondylar repair system for the management of humeral intracondylar fissure and humeral condylar fracture.

OBJECTIVES: To report complications, clinical outcomes and CT-imaging outcomes of a surgical system designed for the management of humeral intracondylar fissures and humeral condylar fractures. MATERIALS AND METHODS: Retrospective review of fracture healing from medical records, direct owner contact and an online data-submission service. Follow-up included CT scans and a calculated "bone-opacity continuity index" to quantify bone healing. RESULTS: There was one major surgical complication and one major medical complication out of 34 fissure cases, and two major surgical and one major medical complication out of 14 fractures. Follow-up times ranged from 29 to 1268 days. All cases with CT follow-up had some continuity of bone opacity across the condyle. CLINICAL SIGNIFICANCE: In the cases included in this study, this repair system was associated with low complication rates and favourable healing rates, particularly for humeral intracondylar fissure.

Systematic review of the management of canine osteoarthritis.

This review assesses the evidence for the efficacy of therapies used in the management of osteoarthritis in dogs on the basis of papers published in peer-reviewed journals in English between 1985 and July 2007. Sixty-eight papers were identified and evaluated. They considered four alternative therapies, one use of functional food, two intra-articular agents, six nutraceutical agents, 21 pharmacological agents, two physical therapies, three surgical techniques and two combinations of weight control. There was a high level of comfort (strong evidence) for the efficacy of carprofen, firocoxib and meloxicam, and a moderate level of comfort for the efficacy of etodolac in modifying the signs of osteoarthritis. There was a moderate level of comfort for the efficacy of glycosaminoglycan polysulphate, licofelone, elk velvet antler and a functional food containing green-lipped mussel for the modification of the structures involved in the disease. There was weak or no evidence in support of the use of doxycycline, electrostimulated acupuncture, extracorporeal shockwave therapy, gold wire acupuncture, hyaluronan, pentosan polysulphate, P54FP (extract of turmeric), tiaprofenic acid or tibial plateau levelling osteotomy.

High-intensity exercise induces structural, compositional and metabolic changes in cuboidal bones--findings from an equine athlete model.

Fatigue fracture of cuboidal bones occurs in the human foot as well as the equine carpus. The racehorse provides a naturally-occurring model to study the effects of high-intensity exercise on the morphology and metabolism of cuboidal bones. We studied both the mineral and the collagenous matrix of the third (C(3)) and radial (C(r)) carpal bones of raced and non-raced Thoroughbred (TB) horses. We hypothesised that racehorses would show increases in the mineral component of these bones and post-translational modifications of the collagenous matrix alongside changes in markers of collagen remodelling and bone formation. C(3) and C(r) carpal bones were retrieved from raced TB horses (n=14) and non-raced TB horses (n=11). Standardised proximal-distal sections were taken from each bone and these were sliced transversely to study the proximal-distal differences in bone metabolism from the subchondral plate through to trabecular bone. Histomorphometry and bone mineral density measurements were performed in parallel with biochemical analyses including total collagen, collagen synthesis and cross-links, matrix metalloproteinases-2 and 9 and their inhibitors, calcium and phosphate, and bone alkaline phosphatase. The results of this study show that, while there is a net increase in bone formation in the racehorses, there is additionally an increase in bone collagen synthesis and remodelling, particularly within the trabecular regions of the bone. The increase in bone density would lead to greater stiffness, particularly in the cortical bone, and failure of this 'stiffer' cortical bone may result from its lack of support from the rapidly remodelling and structurally weakened underlying trabecular bone.

Arthroscopic management of a lateral glenohumeral ligament rupture in two dogs.

Tears to the lateral glenohumeral ligament appear to be uncommon in dogs. Two Labrador Retrievers were arthroscopically diagnosed with lateral glenohumeral ligament tears of the shoulder. Both cases failed to respond to conservative management. Surgical stabilisation of the lateral aspect of the shoulder joint was achieved using an arthroscopically placed suture using a hanging limb technique. Lameness resolved in both cases.

Partial tarsal arthrodesis and botulinum toxin A injection for correction of tarsal arthrogryposis in a cat.

An 11-week-old, domestic shorthair cat was presented for evaluation of a congenital right tarsal deformity. The cat was non-weightbearing lame on the right hindlimb. There was severe tarsal hyperextension and concurrent spasticity/contracture/shortening of the gastrocnemius muscle-tendon. The cat was otherwise healthy. Radiographically there was luxation of the proximal intertarsal joint and talocrural hyperextension. Treatment with 10 mu/kg botulinum toxin A (Botox; Allergan) intramuscular injections directly into the gastrocnemius muscle and passive physiotherapy marginally improved the range of motion in the talocrural joint. Definitive surgical treatment with partial tarsal arthodesis produced an excellent clinical outcome. This case report demonstrates the successful treatment of a severe congenital tarsal abnormality and also documents the first clinical use of botulinum toxin in a feline patient.

Ultrastructural differences in cranial cruciate ligaments from dogs of two breeds with a differing predisposition to ligament degeneration and rupture.

Cranial (anterior) cruciate ligament (CCL) samples were obtained from dogs of the Labrador retriever (LR) and greyhound (GH) breeds, of which the former but not the latter is predisposed to CCL rupture. Electron microscopy revealed that the collagen fibril diameters of GHs were larger than those of LRs (P=0.03). Histological examination revealed a "fibrocartilaginous" appearance of CCLs in seven of eight GHs, and, to a lesser extent, in three of eight LRs. The formation of fibrocartilage is clearly not a disadvantage to the healthy racing GH, and cannot be regarded as a pathological degeneration in this breed. It is suggested that fibrocartilage is formed as a beneficial physiological adaptation to the compression of CCLs caused by tensile stress as a result of the tightening of two twisted bands. Fibrocartilage would appear to protect CCLs in the GH, but it may be indicative of a mild degenerative change, which may eventually lead to rupture in the LR.

Infection of a total hip prosthesis in a dog caused by Achromobacter(Alcaligenes) xylosoxidans.

A four-year-old male, neutered Labrador retriever was presented with progressive left hindlimb lameness 10 months following total hip replacement. Radiography revealed changes consistent with infection and culture of joint fluid from the left coxofemoral joint revealed Achromobacter (Alcaligenes) xylosoxidans. The prosthesis was removed. Culture of the acetabular cup confirmed Achromobacter xylosoxidans. Achromobacter xylosoxidans is an uncommon but serious cause of nosocomial epidemics in hospitals for human beings. To the authors' knowledge, this is the first report of total hip prosthetic infection with Achromobacter xylosoxidans. Little is reported about its pathogenicity in human beings and the authors failed to retrieve any reports of its clinical significance in animals.

Use of biochemical markers of osteoarthritis to investigate the potential disease-modifying effect of tibial plateau levelling osteotomy.

OBJECTIVES: To evaluate the hypothesis that the concentration of the 1/20/5D4 epitope of keratan sulphate, cartilage oligomeric matrix protein and total sulphated glycosaminoglycans in synovial fluids from dogs with cranial cruciate ligament disease would be affected by tibial plateau levelling osteotomy. In addition, to evaluate the hypothesis that medial meniscal release or meniscal injury would alter the expression of these candidate biomarkers. METHODS: Forty-one dogs with naturally occurring cranial cruciate ligament disease were recruited prospectively. Synovial fluids were collected from the index joint before surgery and six weeks and six months postsurgery. Following tibial plateau levelling osteotomy, synovial fluids were assayed for 1/20/5D4 epitope of keratan sulphate and cartilage oligomeric matrix protein concentration using an inhibition ELISA and for sulphated glycosaminoglycans using a direct dye-binding assay. RESULTS: The sulphated glycosaminoglycans ratio did not change significantly during the study. Medial meniscal injury at entry was associated with lower concentrations of synovial fluid cartilage oligomeric matrix protein (P<0.05, unpaired t test). There was no association between medial meniscal release and the changes in marker concentrations, either from 0 to six weeks or 0 to six months. CLINICAL SIGNIFICANCE: Tibial plateau levelling osteotomy did not significantly alter the expression of the named candidate biomarkers. These findings reflect the limited nature of the arthrotomy or indicate that tibial plateau levelling osteotomy does not influence the progression of osteoarthritis (OA). From these studies, there is no evidence that tibial plateau levelling osteotomy affects cartilage metabolism.

Canine mesenchymal stem cells are neurotrophic and angiogenic: An in vitro assessment of their paracrine activity.

Mesenchymal stem cells (MSCs) have been used in cell replacement therapies for connective tissue damage, but also can stimulate wound healing through paracrine activity. In order to further understand the potential use of MSCs to treat dogs with neurological disorders, this study examined the paracrine action of adipose-derived canine MSCs on neuronal and endothelial cell models. The culture-expanded MSCs exhibited a MSC phenotype according to plastic adherence, cell morphology, CD profiling and differentiation potential along mesenchymal lineages. Treating the SH-SY5Y neuronal cell line with serum-free MSC culture-conditioned medium (MSC CM) significantly increased SH-SY5Y cell proliferation (P <0.01), neurite outgrowth (P = 0.0055) and immunopositivity for the neuronal marker ßIII-tubulin (P = 0.0002). Treatment of the EA.hy926 endothelial cell line with MSC CM significantly increased the rate of wound closure in endothelial cell scratch wound assays (P = 0.0409), which was associated with significantly increased endothelial cell proliferation (P <0.05) and migration (P = 0.0001). Furthermore, canine MSC CM induced endothelial tubule formation in EA.hy926 cells in a soluble basement membrane matrix. Hence, this study has demonstrated that adipose-derived canine MSC CM stimulated neuronal and endothelial cells probably through the paracrine activity of MSC-secreted factors. This supports the use of canine MSC transplants or their secreted products in the clinical treatment of dogs with neurological disorders and provides some insight into possible mechanisms of action.

Bilateral fixation of Y-T humeral condyle fractures via medial and lateral approaches in 29 dogs.

OBJECTIVES: To describe bilateral fixation of Y-T fractures of the humeral condyle via combined medial and lateral approaches, and to determine the technique's clinical and radiographic short-term outcomes. METHODS: Details of 30 consecutive fractures in 29 dogs were reviewed. These included signalment, method of fixation, complications, and follow-up limb function and range of elbow joint motion. RESULTS: The age of the dogs ranged from three months to nine years, and bodyweight ranged from 1.9 to 48 kg. The humeral condyle was reattached to the shaft using medial and lateral bone plates in 18 fractures, a medial plate and lateral Kirschner wire(s) in six fractures, and medial and lateral Kirschner wire(s) in six fractures. Major complications were recorded in four fractures and minor complications in two fractures. Limb function at follow-up was graded as excellent in 12, good in 15 and fair in three fractures. The range of elbow flexion was normal in seven, mildly reduced in 18, moderately reduced in four and severely reduced in one fracture. CLINICAL SIGNIFICANCE: In contrast to the caudal approach, combined medial and lateral approaches decrease the extent of periarticular soft tissue dissection, avoid complications associated with olecranon osteotomy and enable exposure of the entire humeral diaphysis for fixation. Bilateral fixation is likely to be better at counteracting bending and torsional forces compared with unilateral fixation.

Mechanical performance in axial compression of a titanium polyaxial locking plate system in a fracture gap model.

OBJECTIVE: To evaluate the bending strength of the VetLOX® polyaxial locking plate system. MATERIALS AND METHODS: Thirty-five 3.5 mm 12-hole titanium VetLOX® plates were used to stabilize seven different construct designs in a 1 cm fracture gap simulation model. Each construct was subjected to axial compression. Mean bending stiffness (BS) and yield load (YL) of each construct design were analysed using a one-way ANOVA and Tukey post-hoc analysis. Screw angulation was measured on reconstructed computed tomography (CT) images. RESULTS: Reducing plate working length for fixed-angle constructs significantly increased BS (p <0.01) and YL (p <0.01). For a constant plate working length, increasing screw number did not significantly affect BS (p = 1.0) or YL (p = 0.86). Screw angulation measurement technique was validated by intra-class correlation coefficients (ICC) (ICC >0.9 for inter- and intra-observer measurements). An average screw angle of 13.2° did not significantly affect mechanical performance although incomplete screw head-plate engagement was noted on some reconstructed CT images when angulation exceeded 10°. Prefabricated screw-head inserts did not significantly increase mechanical performance. A 4 mm bone-plate stand-off distance significantly reduced BS and YL by 63% and 69% respectively. CLINICAL RELEVANCE: The VetLOX® system allows the benefits of polyaxial screw insertion whilst maintaining comparable bending properties to fixed angle insertion. The authors recommend accurate plate contouring to reduce the risk of plate bending.

Ex vivo kinematic studies of a canine unlinked semi-constrained hybrid total elbow arthroplasty system.

OBJECTIVES: Introduction of the Sirius® canine total elbow arthroplasty system, and presentation of the results of a passive range-of-motion analysis based on ex vivo kinematic studies pre-and post-implantation. MATERIALS AND METHODS: Thoracic limbs (n = 4) of medium sized dogs were harvested by forequarter amputation. Plain orthogonal radiographs of each limb were obtained pre- and post-implantation. Limbs were prepared by placement of external fixator pins and Kirschner wires into the humerus and radius. Each limb was secured into a custom-made box frame and retro-reflective markers were placed on the exposed ends of the pins and wires. Each elbow was manually moved through five ranges-of-motion manoeuvres. Data collected included six trials of i) full extension to full flexion and ii) pronation and supination in 90° flexion; a three-dimensional motion capture system was used to collect and analyse the data. The Sirius elbow prosthesis was subsequently implanted and the same measurements were repeated. Data sets were tested for normality. Paired t-tests were used for comparison of pre- and post-implantation motion parameters. RESULTS: Kinematic analysis showed that the range-of-motion (mean and SD) for flexion and extension pre-implantation was 115° ± 6 (range: 25° to 140°). The range-of-motion in the sagittal plane post-implantation was 90° ± 4 (range: 36° to 130°) and this reduction was significant (p = 0.0001). The ranges-of-motion (mean and SD) for supination and pronation at 90° were 50° ± 5, whereas the corresponding mean ranges-of-motion post-implantation were 38° ± 6 (p = 0.0188). CONCLUSION: Compared to a normal elbow, the range-of-motion was reduced. Post-implantation, supination and pronation range-of-motion was significantly reduced at 90° over pre-implantation values. CLINICAL RELEVANCE: These results provide valuable information regarding the effect of the Sirius system on ex vivo kinematics of the normal canine elbow joint. Further, this particular ex vivo model allowed for satisfactory and repeatable kinematic analysis.

Glucose transport and metabolism in chondrocytes: a key to understanding chondrogenesis, skeletal development and cartilage degradation in osteoarthritis.

Despite the recognition that degenerative cartilage disorders like osteoarthritis (OA) and osteochondritis dissecans (OCD) may have nutritional abnormalities at the root of their pathogenesis, balanced dietary supplementation programs have played a secondary role in their management. This review emphasizes the importance and role of nutritional factors such as glucose and glucose-derived sugars (i.e. glucosamine sulfate and vitamin C) in the development, maintenance, repair, and remodeling of cartilage. Chondrocytes, the cells of cartilage, consume glucose as a primary substrate for ATP production in glycolysis and utilize glucosamine sulfate and other sulfated sugars as structural components for extracellular matrix synthesis and are dependent on hexose uptake and delivery to metabolic and biosynthetic pools. Data from several laboratories suggests that chondrocytes express multiple isoforms of the GLUT/SLC2A family of glucose/polyol transporters. These facilitative glucose transporter proteins are expressed in a tissue and cell-specific manner, exhibit distinct kinetic properties, and are developmentally regulated. They may also be regulated by endocrine factors like insulin and insulin-like growth factor I (IGF-I) and cytokines such as interleukin 1 beta (IL-1 beta) and tumour necrosis factor alpha (TNF-alpha). Recent studies suggest that degeneration of cartilage may be triggered by metabolic disorders of glucose balance and that OA occurs coincident with metabolic disease, endocrine dysfunction and diabetes mellitus. Based on these metabolic, endocrine and developmental considerations we present a novel hypothesis regarding the role of glucose transport and metabolism in cartilage physiology and pathophysiology and speculate that supplementation with sugar-derived vitamins and nutraceuticals may benefit patients with degenerative joint disorders.

Pressure profilometry of the lumbosacral disk in dogs.

OBJECTIVE: To investigate the biomechanical behavior of the lumbosacral disk under compressive load in dogs, using pressure profilometry, and to investigate the relationship between pressure profile features and background and disease variables. SAMPLE POPULATION: 23 lumbosacral disks and adjacent vertebrae harvested from medium and large breed dogs. PROCEDURE: A 1.3-mm unidirectional needle-mounted pressure transducer was inserted into the disk in a ventral-to-dorsal manner while the disk was loaded in compression by a materials testing machine. Withdrawal of the transducer resulted in a pressure profile for cranial and lateral stress. Pressure profiles were analyzed, and relationships to age and gross evidence of degeneration were investigated. RESULTS: There was a moderate positive correlation between age and degree of nuclear degeneration (r(s) = 0.420, P = 0.046), but no relationship between age and mean nuclear pressure was detected. Mean nuclear pressure correlated negatively with severity of degenerative changes in the nucleus pulposus. Receiver operator characteristic curves to evaluate mean nuclear pressure as a diagnostic test for nuclear degeneration revealed a sensitivity and specificity of 82 and 83%, respectively. In addition, age was moderately correlated with the magnitude of stress peaks (r(s) = -0.571, P = 0.004). Stress peaks were not related to the severity of nuclear degeneration. CONCLUSIONS AND CLINICAL RELEVANCE: Determination of the mean nuclear pressure by disk profilometry provides information on the severity of lumbosacral disk degeneration with a high degree of sensitivity and specificity. The magnitude of single stress peaks within the dorsal annulus fibrosus is correlated with age and may not necessarily reflect advancing degeneration.

Changes in concentrations of biochemical markers of osteoarthritis following surgical repair of ruptured cranial cruciate ligaments in dogs.

OBJECTIVE: To investigate longitudinal changes in concentrations of the 1/20/5D4 epitope (5D4) of keratan sulfate and total sulfated glycosaminoglycans (S-GAG) in synovial fluid and serum of dogs with cranial cruciate ligament (CCL) rupture that was repaired via intra-articular surgery. ANIMALS: 58 dogs with a ruptured CCL and osteoarthritis of the affected (index) joint. PROCEDURE: Prior to surgical repair of the ruptured CCL, 5D4 concentration was measured in serum and synovial fluid samples by use of an inhibition ELISA, and total S-GAG concentration was measured in synovial fluid samples by use of a direct dye-binding assay. Ruptured CCL were repaired surgically, using an intra-articular fascial graft. Dogs were reexamined 1.5, 7, and 13 months after surgery, and 5D4 and S-GAG concentrations in synovial fluid and serum were measured again. RESULTS: Serum 5D4 concentrations did not change significantly during the study. Concentrations of 5D4 in synovial fluid (expressed as a ratio of S-GAG concentration) did change significantly with time. In the index joint, the 5D4:S-GAG decreased from 0.19 at the beginning of the study to 0.09 1.5 months after surgery, but 7 months after surgery, the ratio increased again to 0.20. CONCLUSIONS AND CLINICAL RELEVANCE: Results support the hypothesis that serum concentration of 5D4 is not a useful marker of osteoarthritis in dogs. Surgical intervention transiently reduced the concentration of 5D4 in synovial fluid but had no effect on S-GAG concentration.

Relations between biochemical markers of osteoarthritis and other disease parameters in a population of dogs with naturally acquired osteoarthritis of the genual joint.

OBJECTIVE: To investigate the hypothesis that concentrations of the keratan sulfate epitope, 1/20/5D4 (5D4) and total sulfated glycosaminoglycans (S-GAG) in synovial fluids, and of 5D4 in serum of dogs with naturally acquired osteoarthritis of the genual joint (stifle), secondary to cranial cruciate ligament deficiency, are associated with other disease parameters of osteoarthritis. ANIMALS: 58 dogs with stifle osteoarthritis secondary to naturally acquired cranial cruciate ligament (CCL) deficiency. PROCEDURE: All dogs were examined clinically, radiographically and, in some instances, scintigraphically. Serum and synovial fluid from both stifles were assayed for 5D4 concentration, using an inhibition ELISA, and for S-GAG, using a direct dye-binding assay. RESULTS: Serum 5D4 concentration was not significantly associated with other disease features. Total S-GAG values were high, and 5D4 values were low, in synovial fluid from clinically active, compared with contralateral joints. The S-GAG concentration in synovial fluid from clinically active joints was negatively correlated with radiographic severity score (rs = -0.389, P = 0.004, Spearman's rank correlation). Other associations between marker concentrations and disease parameters could not be detected. CONCLUSIONS: Serum 5D4 concentration is not a useful marker of stifle osteoarthritis in dogs; however, synovial fluid S-GAG and 5D4 values may hold more promise as disease measures.