Objective—To compare biodegradable magnesium phosphate cement (Mg-cement), calcium phosphate cement (Ca-cement), and no cement on bone repair, biocompatibility, and bone adhesive characteristics in vivo in horses.
Animals—8 clinically normal adulthorses.
Procedures—Triangular fragments (1-cm-long arms) were created by Y-shaped osteotomy of the second and fourth metatarsal bones (MTII and MTIV, respectively). Fragments were replaced in pairs to compare Mg-cement (MTII, n = 8; MTIV, 8) with Ca-cement (MTIV, 8) or with no cement (MTII, 8). Clinical and radiographic evaluations were performed for 7 weeks, at which time osteotomy sites were harvested for computed tomographic measurement of bone density and callus amount, 3-point mechanical testing, and histologic evaluation of healing pattern and biodegradation.
Results—All horses tolerated the procedure without clinical problems. Radiographically, Mg-cement secured fragments significantly closer to parent bone, compared with Ca-cement or no treatment. Callus amount and bone remodeling and healing were significantly greater with Mg-cement, compared with Ca-cement or no cement. Biomechanical testing results and callus density among treatments were not significantly different. Significantly greater woven bone was observed adjacent to the Mg-cement without foreign body reac-tion, compared with Ca-cement or no cement. The Mg-cement was not fully degraded and was still adhered to the fragment.
Conclusions and Clinical Relevance—Both bone cements were biocompatible in horses, and Mg-cementmay assistfracture repair by osteogenesis and fragmentstabilization. Fur ther studies are warranted on other applications and to define degradation characteristics.
Objective—To assess the clinical, biochemical, and histologic effects of intra-articular administration of autologous conditioned serum (ACS) in the treatment of experimentally induced osteoarthritis in horses.
Procedures—Osteoarthritis was induced arthroscopically in 1 middle carpal joint of all horses. In 8 placebo- and 8 ACS-treated horses, 6 mL of PBS solution or 6 mL of ACS was injected into the osteoarthritis-affected joint on days 14, 21, 28, and 35, respectively; PBS solution was administered in the other sham-operated joints. Evaluations included clinical assessment of lameness and synovial fluid analysis (performed biweekly); gross pathologic and histologic examinations of cartilage and synovial membrane samples were performed at necropsy.
Results—No adverse treatment-related events were detected. Horses that were treated with ACS had significant clinical improvement in lameness, unlike the placebo-treated horses. Among the osteoarthritis-affected joints, ACS treatment significantly decreased synovial membrane hyperplasia, compared with placebo-treated joints; although not significant, the ACS-treated joints also appeared to have less gross cartilage fibrillation and synovial membrane hemorrhage. The synovial fluid concentration of interleukin-1 receptor antagonist (assessed by use of mouse anti–interleukin-1 receptor antagonist antibody) was increased following treatment with ACS.
Conclusions and Clinical Relevance—Results of this controlled study indicated that there was significant clinical and histologic improvement in osteoarthritis-affected joints of horses following treatment with ACS, compared with placebo treatment. On the basis of these findings, further controlled clinical trials to assess this treatment are warranted, and investigation of the mechanisms of action of ACS should be pursued concurrently.
Objective—To evaluate anticollagen type I antibodies in synovial fluid of the affected stifle joint, the contralateral stifle joint, and the left shoulder joint of dogs with unilateral cranial cruciate ligament (CrCL) rupture during an extended period of 12 to 18 months.
Animals—13 client-owned dogs with CrCL rupture and 2 sham-operated dogs.
Procedures—All dogs were examined and arthrocentesis of all 3 joints was performed every 6 months after surgery. Synovial fluid samples were tested for anticollagen type I antibodies by use of an ELISA.
Results—Dogs with partial CrCL rupture had higher antibody titers than dogs with complete rupture. Six of 13 dogs ruptured the contralateral CrCL during the study, whereby higher antibody titers were found for the stifle joints than for the shoulder joint. Seronegative dogs or dogs with extremely low antibody titers and 2 dogs with high antibody titers did not sustain a CrCL rupture in the contralateral stifle joint.
Conclusions and Clinical Relevance—In most dogs that had a CrCL rupture of the contralateral stifle joint, a distinct antibody titer gradient toward the stifle joints was detected, suggesting that there was a local inflammatory process in these joints. However, only a small number of sham-operated dogs were used to calculate the cutoff values used to determine the anticollagen type I antibody titers in these patients. Synovial fluid antibodies against collagen type I alone do not initiate CrCL rupture because not all dogs with high antibody titers sustained a CrCL rupture in the contralateral stifle joint.
Objective—To evaluate changes in serum cartilage oligomeric matrix protein (COMP) concentrations in response to exercise in horses.
Animals—15 horses in experiment 1 and 27 horses in experiment 2.
Procedures—In experiment 1, 15 Thoroughbreds free of orthopedic disease underwent a standardized exercise protocol. Running velocity and heart rate (HR) were recorded, and blood samples were collected immediately before (baseline) and 1, 5, and 24 hours after a single episode of exercise. In experiment 2, 27 horses underwent 9 stages of a training program in which each stage consisted of 4 to 8 consecutive daily workouts followed by a rest day. Blood samples were collected immediately before the first and final daily workouts in each stage. Serum COMP concentrations were measured via inhibition ELISA with a monoclonal antibody (14G4) against equine COMP.
Results—In experiment 1, mean serum COMP concentration was significantly higher than baseline 1 and 5 hours after exercise and returned to baseline concentrations 24 hours after exercise. Mean serum baseline COMP concentration increased as the velocity of running at maximum HR and at an HR of 200 beats/min increased, being significantly higher during the third and fourth exercise tests than during the first. In experiment 2, mean baseline COMP concentration at the final workout of each stage was significantly higher than that at the first workout, beginning with stage 3.
Conclusions and Clinical Relevance—Serum COMP concentrations changed significantly in response to exercise. Exercise may enhance movement of COMP into the circulation as well as change the basal turnover rate of COMP.
Objective—To evaluate the effects of continuous oral administration of phenylbutazone on serum and synovial fluid biomarkers of skeletal matrix metabolism in horses.
Animals—11 adult female horses without clinical or radiographic evidence of joint disease.
Procedures—Horses were randomly assigned to control or treatment groups. Phenylbutazone was administered orally twice daily at a dose of 4.4 mg/kg for 3 days to the treatment group and subsequently at a dose of 2.2 mg/kg for 7 days. Serum and radiocarpal synovial fluid samples were obtained at baseline and thereafter at regular intervals for 4 weeks. Biomarkers of cartilage aggrecan synthesis (chondroitin sulfate 846) and type II collagen synthesis (procollagen type II C-propeptide) and degradation (collagen type II cleavage) were assayed. Biomarkers of bone synthesis (osteocalcin) and resorption (C-terminal telopeptide of type I collagen) were also measured.
Results—No significant differences were found between control and treatment groups or temporally for the biomarkers chondroitin sulfate 846, procollagen type II C-propeptide, collagen type II cleavage, and C-terminal telopeptide of type I collagen in serum or synovial fluid. A significant increase in osteocalcin concentration occurred in synovial fluid during treatment in the treated group. No treatment effect was detected for serum osteocalcin concentration.
Conclusions and Clinical Relevance—Results suggested that continuous phenylbutazone administration at recommended doses altered some biomarkers in healthy equine joints after short periods of administration. Increased osteocalcin concentration may indicate an undetermined anabolic effect of phenylbutazone administration on periarticular bone or transient induction of osteogenesis in articular chondrocytes or a mesenchymal subpopulation of synoviocytes.
Objective—To evaluate the quantitative inheritance of secondary hip joint osteoarthritis in a canine pedigree.
Animals—137 Labrador Retrievers, Greyhounds, and mixed-breed dogs.
Procedures—Necropsy scores ranging from 0 to 4 were obtained for each hip joint. Seven unaffected Greyhounds with normal hip joint conformation were also used for genetic modeling, but were not euthanized. Sixty-six male and 71 female dogs were allocated to 2 groups (≤ 12 months of age and > 12 months of age). Statistical models were developed to establish the inheritance pattern of hip joint osteoarthritis that developed secondary to hip dysplasia.
Results—62 dogs had evidence of osteoarthritis in a hip joint, and 75 had no evidence of osteoarthritis. After sex was adjusted for, the necropsy score was found to be inherited additively but without dominance. Each Labrador Retriever allele increased the necropsy score by 0.7 to 0.9 points, compared with the Greyhound allele, and male sex increased the necropsy score 0.74 over female sex. Approximately 10% of the variation in necropsy score was attributable to the litter of puppies' origin.
Conclusions and Clinical Relevance—Because secondary hip joint osteoarthritis is inherited additively, selection pressure could be applied to reduce its incidence. Similar statistical models can be used in linkage and association mapping to detect the genes in the underlying quantitative trait loci that contribute to hip joint osteoarthritis.
Objective—To compare measurements obtained by use of a universal plastic goniometer (UG) and an electrogoniometer (EG) and from radiographs and to compare joint motion in German Shepherd Dogs and Labrador Retrievers.
Animals—12 healthy adult German Shepherd Dogs and data previously collected from 16 healthy adult Labrador Retrievers.
Procedures—German Shepherd Dogs were sedated. One investigator then measured motion of the carpal, cubital (elbow), shoulder, tarsal, stifle, and hip joints of the sedated dogs. Measurements were made in triplicate with a UG and an EG. Radiographs were taken of all joints in maximal flexion and extension. Values were compared between the UG and EG and with values previously determined for joints of 16 Labrador Retrievers.
Results—An EG had higher variability than a UG for all dogs. The EG variability appeared to result from the technique for the EG. German Shepherd Dogs had lower values in flexion and extension than did Labrador Retrievers for all joints, except the carpal joints. German Shepherd Dogs had less motion in the tarsal joints, compared with motion for the Labrador Retrievers, but had similar motion in all other joints.
Conclusions and Clinical Relevance—A UG is reliable for obtaining measurements in German Shepherd Dogs. There was higher variability for the EG than for the UG, and an EG cannot be recommended for use.
Objective—To evaluate the involvement of various collagen genes in the development of fragmented medial coronoid process (FCP) in Labrador Retrievers.
Sample Population—93 dogs originating from 13 litters were used in the study; FCP was diagnosed in 35 dogs, and each affected dog had at least 1 sibling that was also affected. Twelve dams and sires were included in the analysis. All dogs were purebred Labrador Retrievers except for 2 litters (offspring of a female Golden Retriever-Labrador Retriever mixed-breed dog).
Procedures—For each dog, DNA was isolated from blood samples. Polymorphic microsatellite markers adjacent to 14 candidate genes (ie, COL1A1, COL1A2, COL2A1, COL3A1, COL5A1, COL5A2, COL6A3, COL9A1, COL9A2, COL9A3, COL10A1, COL11A1, COL11A2, and COL24A1) were analyzed by use of PCR assays; genotypes were determined via automated detection of DNA products. The level of allele sharing between pairs of affected siblings was assessed.
Results—Among the 93 dogs, allele sharing of the 14 collagen genes was determined as follows: COL1A1, 45%; COL1A2, 47%; COL2A1, 37%; COL3A1, 32%; COL5A1, 43%; COL5A2, 32%; COL6A3, 36%; COL9A1, 45%; COL9A2, 49%; COL9A3, 38%; COL10A1, 46%; COL11A1, 52%; COL11A2, 47%; and COL24A1, 47%.
Conclusions and Clinical Relevance—Because siblings share 50% of their genome at random, the fact that the percentages of allele sharing among the analyzed collagen genes were not significantly > 50% indicates that these genes are not determinant candidates for FCP in Labrador Retrievers. The gene for the vitamin D receptor could also be excluded because of its proximity to COL2A1.
Objective—To determine the critical temperature that reduces chondrocyte viability and evaluate the ability of chondrocytes to recover after exposure to the critical temperature.
Sample Population—Cartilage explants obtained from the humeral heads of 30 sheep.
Procedures—In a randomized block design, 318 full-thickness cartilage explants were collected from 30 humeral heads of sheep and cultured for up to 14 days. On the first day of culture (day 0), explants were subjected to temperatures of 37°, 45°, 50°, 55°, 60°, or 65°C for 5 minutes by heating culture tubes in a warming block. The ability for chondrocytes to recover after exposure to the critical temperature was determined by evaluating viability at days 0, 1, 3, 7, and 14 days after heating. Images were analyzed by use of confocal laser microscopy.
Results—Analysis of images revealed a significant decrease in live cells and a significant increase in dead cells as temperature increased. Additionally, the deepest layer of cartilage had a significantly lower percentage of live cells, compared with values for the 3 most superficial layers. Chondrocytes did have some ability to recover temporarily after the initial thermal insult.
Conclusions and Clinical Relevance—A strong relationship exists between increasing temperature and cell death, with a sharp increase in chondrocyte death between 50° and 55°C. Chondrocytes in the deepest cartilage layer are most susceptible to thermal injury. The threshold of chondrocyte recovery from thermal injury is much lower than temperatures reached during chondroplasty by use of most radiofrequency energy devices.
Objective—To quantify and compare the microscopic changes in articular cartilage (AC), zone of calcified cartilage (ZCC), and subchondral bone plate in femoral heads from clinically normal dogs and dogs with moderate or severe osteoarthritis.
Sample Population—Femoral heads from clinically normal dogs (n = 16) and dogs with moderate (24) or severe (14) osteoarthritis.
Procedures—Femoral heads were allocated to 3 categories (normal, moderate, or severe osteoarthritis) on the basis of radiographic findings, macroscopic findings, and histologic grade determined by use of a modified Mankin scale. Equally spaced 2-mm sections were cut in each femoral head in a coronal or transverse plane. Thickness of the AC, ZCC, and subchondral bone plate was recorded.
Results—Mean thickness of AC was significantly greater in samples with moderate and severe osteoarthritis than those considered normal. Mean thickness of the ZCC was significantly greater in samples with moderate and severe osteoarthritis than those considered normal. Mean thickness of the subchondral bone plate in samples with severe osteoarthritis was significantly greater than those with moderate osteoarthritis and those considered normal. A significant decrease in AC thickness was detected in the proximomedial area of femoral heads with severe osteoarthritis, compared with those considered normal.
Conclusions and Clinical Relevance—A cause and effect association between thickening of subchondral structures and thinning and loss of the overlying AC was not detected. Changes in AC were associated with changes in the subchondral bone plate, which is compatible with the theory of adaptation in response to altered load distribution.