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Abstract

Objective—To determine whether a single contusive impact injury to the palmar aspect of the metacarpus would progress to post-traumatic osteoarthritis or palmar osteochondral disease in horses.

Animals—12 horses.

Procedures—In each horse, an impact injury was created on the palmar aspect of the medial metacarpal condyle of 1 randomly chosen limb with an impactor device under arthroscopic and fluoroscopic guidance. The opposite limb was sham operated as a control. A low to moderate amount of forced exercise was instituted, and horses were evaluated clinically via lameness examinations weekly for 5 months, then biweekly until endpoint, with synovial fluid analysis performed at 0, 1, 2, 3, 4, 6, 8, and 10 months and radiography at baseline and endpoint. Macroscopic examination, micro-CT, and sample collection for cartilage viability and sulfated glycosaminoglycan content, histologic evaluation, immunohistochemical analysis, and fluorochrome analysis were performed following euthanasia at 1 (3 horses), 4 (4), and 8 to 10 (5) months after surgery.

Results—There was variability in impact lesion location, depth, and area on macroscopic inspection, but on histologic evaluation, cartilage defects were less variable. Mean sulfated glycosaminoglycan concentration from cartilage at the impact site was significantly lower than that at a similar site in control limbs. Higher concentrations of cartilage oligomeric matrix protein were observed in synovial fluid from impact-injured joints.

Conclusions and Clinical Relevance—The impact injury method caused mild focal osteoarthritic lesions in the metacarpophalangeal joint, but did not progress to palmar osteochondral disease at this site. Repeated injury is probably required for the development of palmar osteochondral disease.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To investigate the relationship between runt-related transcription factor 2 (RUNX2) expression in canine nucleus pulposus (NP) cells and intervertebral disk aging in chondrodystrophoid dogs.

Animals—7 healthy Beagles (mean age, 35.6 months) and 11 Dachshunds with herniated disks (mean age, 61 months).

Procedures—All dogs underwent MRI examination of the thoracic and lumbar vertebral column immediately before sample collection under general anesthesia. The disk center–to–CSF T2-weighted signal intensity ratio was determined for healthy Beagles. Samples of NP were obtained from nonherniated disks in healthy Beagles and from herniated disks during surgical treatment of hospitalized Dachshunds. Samples were evaluated for RUNX2 and matrix metalloproteinase 13 transcript expression via reverse transcriptase PCR assay; RUNX2 protein expression was evaluated via immunohistochemical analysis, and correlation between these variables and age of dogs was evaluated. A 3′ and 5′ rapid amplification of cDNA ends method was used to identify the RUNX2 coding region.

Results—RUNX2 cDNA had > 97% conservation with the human cDNA sequence and approximately 95% conservation with the mouse cDNA sequence; RUNX2 and matrix metalloproteinase 13 mRNA expression and RUNX2 protein expression in NP cells were positively correlated with age. The disk center–to–CSF T2-weighted signal intensity ratio was negatively correlated with RUNX2 protein expression in the NP of healthy dogs.

Conclusions and Clinical Relevance—Results indicated that RUNX2 mRNA and protein expression in the NP are enhanced in aging intervertebral disks in dogs.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate transduction efficiency of gene therapy for treatment of osteoarthritis in horses.

Sample—Cartilage and synovial tissues were aseptically collected from the stifle joints of 3 Thoroughbreds; horses were 3, 7, and 12 years old and free from sepsis and long-term drug treatment and were euthanized for reasons unrelated to joint disease.

Procedures—Gene transfer experiments were performed with 8 recombinant adeno-associated viral vector (rAAV) serotypes in monolayer-cultured equine chondrocytes, synovial cells, and mesenchymal stromal cells and in cartilage and synovial tissues.

Results—Serotypes rAAV2/5 and rAAV2/2 yielded the highest transduction efficiency in cultured cells 6 days after transduction. Synovial cells and mesenchymal stromal cells were more readily transduced than were chondrocytes. Serotype rAAV2/6.2 yielded the highest rate of gene expression in both cartilage and synovial tissues at 6 days after inoculation. However, at 30 and 60 days after inoculation, gene expression of serotypes rAAV2/2 and rAAV2/5 surpassed that of rAAV2/6.2 and all other serotypes.

Conclusions and Clinical Relevance—Maximally expressing serotypes changed between 6 and 30 days in tissues; however, the most efficient serotypes for transduction of joint cells over time were also the most efficient serotypes for transduction of joint tissues. In addition, the low transduction efficiency of articular cartilage tissue was paralleled by a low transduction efficiency of isolated chondrocytes. This suggested that the typically low transduction efficiency of articular cartilage may be attributable in part to the low transduction efficiency of the chondrocytes and not solely a result of the dense cartilage matrix.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To investigate the effects of gonadectomy on collagen homeostasis in cranial cruciate ligaments of male rabbits.

Animals—30 sexually immature (16-week-old) male New Zealand White rabbits.

Procedures—Rabbits were randomly assigned to 5 groups of 6 rabbits each: sexually intact, placebo (control group); castrated, placebo; castrated, testosterone; castrated, dihydrotestosterone; and castrated, 17β-estradiol (E2). Control rabbits underwent a sham operation, and all other rabbits underwent gonadectomy. At the time of gonadectomy, the placebo and sex hormones were administered via slow-release pellets implanted subcutaneously as assigned. After 21 days of hormone supplementation, measurements were obtained of serum testosterone and E2 concentrations, ligament collagen characteristics, and androgen receptor, estrogen receoptor α, and matrix metalloproteinase expression.

Results—Following gonadectomy and hormone supplementation, the treatment groups differed in serum testosterone and E2 concentrations to various degrees. Collagen concentrations were lower and fiber diameters higher in the absence of sex hormones, in association with the degrees of estrogen receptor a and androgen receptor expression. Although differences were detected among the groups in matrix metalloproteinase expression, these differences were not significant.

Conclusions and Clinical Relevance—Sex hormones appeared to play a role in cranial cruciate ligament homeostasis in male rabbits. Physiologic changes triggered by the lack of sex hormones following gonadectomy in sexually immature rabbits may potentially predispose those rabbits to orthopedic injuries.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To assess clinical, radiographic, histologic, and biochemical effects of sodium pentosan polysulfate (NaPPS) administered IM for treatment of experimentally induced osteoarthritis in horses.

Animals—18 horses.

Procedures—Osteoarthritis was induced arthroscopically in 1 middle carpal joint of all horses. Nine horses received NaPPS (3 mg/kg, IM) on study days 15, 22, 29, and 36. Nine control horses received the same volume of saline (0.9% NaCl) solution IM on study days 15, 22, 29, and 36. Clinical, radiographic, gross, histologic, histochemical, and biochemical findings as well as findings of synovial fluid analysis were evaluated.

Results—No adverse treatment-related events were detected. Induced osteoarthritis caused a substantial increase in lameness, response to flexion, joint effusion, radiographic findings, synovial membrane inflammation, and articular cartilage fibrillation. Articular cartilage fibrillation was substantially reduced by NaPPS treatment, and concentrations of chondroitin sulfate 846 epitope were significantly increased in the synovial fluid of osteoarthritic and nonosteoarthritic joints of treated horses.

Conclusions and Clinical Relevance—Results indicated that NaPPS has some beneficial disease-modifying effects and may be a therapeutic option for osteoarthritis in horses.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To describe and measure histologic features of midcarpal joint cartilage defects in Thoroughbreds and evaluate the influence of early conditioning exercise on defect development.

Sample—24 midcarpal joints from twelve 18-month-old Thoroughbreds.

Procedures—Midcarpal joints from 12 horses (6 exercised spontaneously at pasture only and 6 given additional conditioning exercise beginning at a mean age of 3 weeks were evaluated. Gross cartilage defects were assessed histologically. Third and radial carpal bones were categorized with regard to the presence or absence of calcified cartilage (CC) abnormalities at the dorsoproximal and dorsodistal articular surfaces, respectively; histomorphometric assessment and statistical analysis were conducted for the third carpal bone.

Results—Number and severity of defects did not appear different between exercise groups. Nine third or radial carpal bones had thickened CC with microcracks, matrix and osteochondral junction changes, and increased vascularity, without histologic changes in the hyaline cartilage. Third carpal bones with CC abnormalities had significantly thicker CC (452 vs 228 μm) than did those without CC abnormalities in the evaluated region. However, in the same region, there were no significant differences in hyaline cartilage thickness (681 vs 603 μm), vascular channel area in the subchondral bone (624,894 vs 490,320 μm2), or number of vascular channels (15.9 vs 18.0).

Conclusions and Clinical Relevance—Early exercise did not appear to influence the distribution or severity of cartilage defects in the midcarpal joint. Calcified cartilage abnormalities beneath the undisrupted hyaline cartilage in the dorsoproximal aspect of the third carpal bone may represent the first changes in the pathogenesis of midcarpal osteochondral disease.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate and compare bone modeling and remodeling in fractured and non-fractured central tarsal bones (CTBs) of racing Greyhounds.

Sample—Paired cadaveric tarsi from 6 euthanized racing Greyhounds with right CTB fractures and 6 racing Greyhounds with other nontarsal injuries.

Procedures—CTBs were dissected and fractured CTBs were reconstructed. Central tarsal bones were evaluated through standard and nonscreen high-detail radiography, computed tomography, and histologic examination. The bone mineral density (BMD) was calculated adjacent to fracture planes and as a gradient on sagittal computed tomographic images. Sagittal and transverse plane sections of bone were obtained and submitted for subjective histologic assessment. Linear mixed-effects models were used to compare findings.

Results—Fractured right CTBs had greater BMD in the dorsal and midbody regions of the sagittal plane sections than did nonfractured CTBs. The BMD ratios from bone adjacent to the dorsal slab fracture planes were not different between fractured and nonfractured right CTBs.

Conclusions and Clinical Relevance—Findings supported the existence of site-specific bone adaptation in CTBs of Greyhounds, with modeling and remodeling patterns that were unique to fractured right CTBs. The dorsal and midbody regions of fractured bones had greater BMD, and fractures occurred through these zones of increased BMD.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To investigate the relationship between inflammatory responses of the temporomandibular joint (TMJ) and the metacarpophalangeal (MCP) joint in clinically normal horses.

Animals—7 mature horses.

Procedures—In each horse, 1 TMJ and 1 MCP joint were injected with lipopolysaccharide (LPS; 0.0025 μg). The contralateral TMJ and MCP joint were injected with saline (0.9% NaCl) solution. Synovial fluid samples were collected from all 4 joints over 24 hours after injection. Concentrations of interleukin-6, tumor necrosis factor-α, transforming growth factor-β, and total protein were measured via immunoassay. Horses were assessed for clinical signs of joint inflammation at each time point.

Results—Concentrations of interleukin-6 were not significantly different between LPS-injected MCP joints and TMJs at any time point. Transforming growth factor-β concentrations were significantly increased in MCP joints, compared with concentrations in TMJs, at 12 and 24 hours after injection. Tumor necrosis factor-α concentrations were significantly higher in LPS-injected TMJs than in LPS-injected MCP joints at 1 and 6 hours after injection. Total protein concentration did not differ significantly between LPS-injected MCP joints and TMJs. Injection of LPS induced clinical inflammation at all time points; additionally, 2 MCP joints (but no TMJs) had an inflammatory response to injection of saline solution.

Conclusions and Clinical Relevance—The inflammatory response to LPS appeared to be attenuated more quickly in TMJs than in MCP joints of horses. The difference in response suggested that a lack of clinical osteoarthritis in the TMJ of horses could be attributable to a difference in cytokine response.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate cartilage thickness of the talus (especially at sites predisposed to osteochondrosis dissecans [OCD]) in growing and adult dogs not affected with OCD.

Sample—Tarsocrural joints from cadavers of 34 juvenile (approx 3 months old) and 10 adult dogs.

Procedures—Tarsal cartilage thickness was examined via a stereophotography microscopic system. Articular cartilage thickness was determined at 11 locations on longitudinal slices of the trochlear ridges and the sulcus between the ridges and at 2 locations in the cochlea tibiae. Cartilage thickness was measured at the proximal, proximodorsal, dorsal, and distal aspects of the trochlear ridges; proximodorsal, dorsal, and distal aspects of the trochlear sulcus; and craniolateral and caudomedial aspects of the cochlea tibiae. Differences within a joint and between sexes were evaluated.

Results—Mean cartilage thickness decreased from proximal to distal in juvenile (lateral trochlear ridge, 1.52 to 0.41 mm; medial trochlear ridge, 1.10 to 0.40 mm) and from proximal to dorsal in adult (lateral trochlear ridge, 0.41 to 0.34 mm; medial trochlear ridge, 0.33 to 0.23 mm) dogs. Cartilage was thickest at the proximal aspect of the lateral trochlear ridge in both groups. Differences in proximodorsal, dorsal, and distal aspects of the ridges were not evident.

Conclusions and Clinical Relevance—Healthy tarsocrural joints did not have thicker cartilage in sites predisposed to development of OCD. Evaluation of affected tarsocrural joints is necessary to exclude influences of cartilage thickness. These data are useful as a reference for distribution of cartilage thickness of the trochlea of the talus in dogs.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To assess the effect of computed tomography (CT) scan protocols (radiation amounts) and fabrication methods on biomodel accuracy and variability.

Sample—Cadaveric femur of a Basset Hound.

Procedures—Retroreconstructions (n = 158) were performed of 16 original scans and were visually inspected to select 17 scans to be used for biomodel fabrication. Biomodels of the 17 scans were made in triplicate by use of 3 freeform fabrication processes (stereolithography, fused deposition modeling, and 3-D printing) for 153 models. The biomodels and original bone were measured by use of a coordinate measurement machine.

Results—Differences among fabrication methods accounted for 2% to 29% of the total observed variation in inaccuracy and differences among method-specific radiation configurations accounted for 4% to 44%. Biomodels underestimated bone length and width and femoral head diameter and overestimated cortical thickness. There was no evidence of a linear association between thresholding adjustments and biomodel accuracy. Higher measured radiation dose led to a decrease in absolute relative error for biomodel diameter and for 4 of 8 cortical thickness measurements.

Conclusions and Clinical Relevance—The outside dimensions of biomodels have a clinically acceptable accuracy. The cortical thickness of biomodels may overestimate cortical thickness. Variability among biomodels was caused by model fabrication reproducibility and, to a lesser extent, by the radiation settings of the CT scan and differences among fabrication methods.

Full access
in American Journal of Veterinary Research