Technical note: Use of marker-based relationships with multiple-trait derivative-free restricted maximal likelihood.

The widespread use of the set of multiple-trait derivative-free REML programs for prediction of breeding values and estimation of variance components has led to significant improvement in traits of economic importance. The initial version of this software package, however, was generally limited to pedigree-based relationships. With continued advances in genomic research and the increased availability of genotyping, relationships based on molecular markers are obtainable and desirable. The addition of a new program to the set of multiple-trait derivative-free REML programs is described that allows users the flexibility to calculate relationships using standard pedigree files or an arbitrary relationship matrix based on genetic marker information. The strategy behind this modification and its design is described. An application is illustrated in a QTL association study for canine hip dysplasia.

Analysis of allele fidelity, polymorphic information content, and density of microsatellites in a genome-wide screening for hip dysplasia in a crossbreed pedigree.

Recent advances in genomics resources and tools are facilitating quantitative trait locus mapping. We developed a crossbreed pedigree for mapping quantitative trait loci for hip dysplasia in dogs by crossing dysplastic Labrador Retrievers and normal Greyhounds. We show that one advantage to using a crossbreed pedigree is the increased marker informativeness in the backcross/F2 population relative to the founder populations. We also discuss three factors that affect the detection power in the context of this crossbreed pedigree: being able to detect and correct genotyping errors, increasing marker density for chromosomes with a sparse coverage, and adding individuals to the mapping population as soon as they become available.

Genes in canine articular cartilage that respond to mechanical injury: gene expression studies with Affymetrix canine GeneChip.

The Affymetrix canine GeneChip with 23,836 probe sets was used to look for cartilage genes that are significantly altered in response to mechanical impact. The model using canine articular cartilage explants loaded in vitro has been described previously (Chen et al., J Orthop Res 19:703-711, 2001). It is our hypothesis that genes that are activated or repressed in articular cartilage after impact injury initiate cartilage degeneration, leading to osteoarthritis in dogs. Gene expression of known cartilage genes was generally consistent with cartilage biology. A total of 528 genes were significantly (P < .01) up- or down- regulated in response to mechanical damage. After applying the strict Bonferroni correction, 172 remained significantly affected. One of these genes, MIG-6/gene 33, was chosen for verification by real- time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). A 3.8- fold increase in expression was confirmed, consistent with the microarray chip data. Deficiencies in the current annotation of the canine chip are discussed. Gene expression studies with the Affymetrix canine GeneChip are potentially valuable, but await more complete annotation.

In vitro biomechanical comparison of limited contat dynamic compression plate and locking compression plate.

The locking compression plate (LCP) supports biological osteosynthesis by functioning as an internal fixator, rather than as a full or limited contact bone plate which must be adequately contoured and affixed directly to the bone for stable internal fixation of the fracture. In order to help justify the use of the LCP in our veterinary patients, in vitro biomechanical testing was performed comparing the LCP to the conventional limited contact dynamic compression plate (LC-DCP) in canine femurs. We hypothesized that the LCP construct would be at least as stiff under bending and torsional loads as the LC-DCP. The LCP and LC-DCP were applied over a 20-mm osteotomy gap to contralateroal bones within each pair of 14 femora. Non-destructive four-point bending and torsion, and cyclical testing in torsion were performed. The constructs were then loaded to failure in torsion. In medial-lateral and lateral-medial structural bending, significant differences were not found between the LCP and LC-DCP, however, at the gap, the LCP construct was stiffer than the LC-DCP in lateral-medial bending. Significant differences in behaviour over time were not noted between the plate designs during cyclical testing. When loading the constructs to failure in internal rotation, the LC-DCP failed at a significantly lower twist angle (P = .0024) than the LCP. Based on the similar performance with loading, the locking compression plate is a good alternative implant for unstable diaphyseal femoral fracture repair in dogs.

Genetic structure of susceptibility traits for hip dysplasia and microsatellite informativeness of an outcrossed canine pedigree.

An outcrossed canine pedigree was developed for quantitative trait locus (QTL) mapping of hip dysplasia by breeding dysplastic Labrador retrievers to trait-free greyhounds. Measured susceptibility traits included age at onset of femoral capital chondroepiphyseal ossification (OSS), maximum hip distraction (laxity) index (DI), and the dorsolateral subluxation (DLS) score. The pedigrees consisted of 147 dogs representing four generations. For 59 dogs genotyped with 65 microsatellites, the median heterozygosity and polymorphic information content (PIC) values of the F(1) generation were 0.82 and 0.68, respectively. Seventy-seven percent of microsatellites had a PIC greater than 0.59 in the F(1)s. Ninety-six percent of alleles showed Mendelian inheritance. Based on marker informativeness, approximately 350 randomly selected markers would be required for genome-wide screening to obtain an average interval between informative markers of 10 cM. Heritability was estimated as 0.43, 0.5, and 0.61 for OSS, DI, and the DLS score, respectively. Biometric estimates of the mean (+/- variance) effective number of segregating QTLs was 1.2 (+/- 0.05), 0.8 (+/- 0.02), and 1.0 (+/- 0.03) for OSS, DI, and the DLS score, respectively. The distributions of simulated backcross trait data suggested that the loci controlling these traits acted additively and that the DI may be controlled by a major locus. When combined with previous power and quantitative genetic analyses, these estimates indicate that this pedigree is informative for QTL mapping of hip dysplasia traits.

Repeatability of dorsolateral subluxation scores in dogs and correlation with macroscopic appearance of hip osteoarthritis.

OBJECTIVE: To determine whether dorsolateral subluxation (DLS) scores in young dogs could be used to reliably predict which dogs would develop evidence of hip osteoarthritis and whether DLS scores measured at various ages correlated with each other. ANIMALS: 129 Labrador Retrievers, Greyhounds, and Labrador Retriever-Greyhound crossbreds. PROCEDURES: DLS scores were measured on radiographs taken at 4, 8, and 12 months of age and at necropsy (8 to 36 months of age). At necropsy, the hip joints were examined macroscopically and a score assigned for degree of cartilage degeneration. RESULTS: DLS scores at 4 (n = 35, r(s) = -0.62), 8 (n = 106, r(s) = -0.54), and 12 (n = 15, r(s) = -0.87) months of age were significantly correlated with cartilage degeneration scores, and DLS scores at 8 months of age were significantly correlated with scores obtained at the time of necropsy (n = 39, r(s) = 0.87). The DLS scores at 4 months of age were significantly different from scores at 8 months of age, but scores did not differ significantly thereafter. Likelihood ratios for cartilage lesions for low (< 45%), intermediate (> or = 45 but < or = 55%), and high (> 55%) DLS scores at 8 months of age were 8.0, 2.6, and 0.2, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that DLS score at 8 months of age was a reasonable, albeit imperfect, predictor of the condition of the hip joint cartilage at necropsy. Thus, the DLS method might be useful for early identification of dogs with hip dysplasia.

Comparison of three radiographic methods for diagnosis of hip dysplasia in eight-month-old dogs.

OBJECTIVE: To compare the accuracy of the extended-hip radiographic (EHR) score, the distraction index (DI), and the dorsolateral subluxation (DLS) score for identifying hip dysplasia in dogs at 8 months of age. DESIGN: Cohort study ANIMALS: 129 Labrador Retrievers, Greyhounds, and Labrador Retriever-Greyhound crossbreds. PROCEDURE: Radiography was performed when dogs were 8 months of age. Dogs were euthanatized at 8 to 36 months of age; hip dysplasia was diagnosed at the time of necropsy on the basis of results of a gross examination of the articular cartilage of the hip joints for signs of osteoarthritis. RESULTS: The EHR score, DI, and DLS score at 8 months of age were all significantly correlated with degree of cartilage degeneration at necropsy. Sensitivity and specificity of using EHR score at 8 months of age to diagnose hip dysplasia (scores > 3 were considered abnormal) were 38 and 96%, respectively; sensitivity and specificity of using DI (values > 0.7 were considered abnormal) were 50 and 89%; and sensitivity and specificity of using DLS score (scores < 55% were considered abnormal) were 83 and 84%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that specificities of the 3 methods for diagnosing hip dysplasia in dogs at 8 months of age were similar. However, the DLS score had higher sensitivity, indicating that there were fewer false-negative results.

Corticosteroids alter the differentiated phenotype of articular chondrocytes.

Experimental evidence suggests that recommended dosages of some corticosteroids used clinically as antiinflammatory agents for treating arthropathies damage articular cartilage, but low dosages may be chondroprotective. The purpose of this study was to evaluate how different concentrations of methylprednisolone affect chondrocyte function and viability. Articular cartilage and chondrocytes were obtained from young adult horses, 1.5-3.5 years of age. Corticosteroid-induced changes in collagen expression were studied at the transcriptional level by Northern blot analyses and at the translational level by measuring [3H]-proline incorporation into [3H]-hydroxyproline. Fibronectin mRNA splicing patterns were evaluated with ribonuclease protection assays. Cytotoxicity was studied using erythrosin B dye exclusion. Steady-state levels of type II procollagen mRNA decreased without concurrent changes in type I procollagen expression as the medium methylprednisolone concentrations were increased from 1 x 10(1) to 1 x 10(8) pg/ml, dropping below 10% of control values by 1 x 10(5) pg/ml. Cytotoxicity occurred as methylprednisolone levels were increased further from 1 x 10(8) to 1 x 10(9) pg/ml. Changes in total collagen (protein) synthesis were less pronounced, but also demonstrated significant suppression between 1 x 10(4) and 1 x 10(8) pg/ml. Corticosteroid-induced changes in fibronectin isoform levels were evaluated in articular cartilage samples without in vitro culture. The cartilage-specific (V + C)(-) isoform was suppressed in both normal and inflamed joints by a single intraarticular injection (0.1 mg/kg) of methylprednisolone. Combined, these data indicate that methylprednisolone suppresses matrix protein markers of chondrocytic differentiation. Decreased and altered chondrocyte expression of matrix proteins likely contributes to the pathogenesis of corticosteroid-induced cartilage degeneration.

The effects of methylprednisolone on normal and monocyte-conditioned medium-treated articular cartilage from dogs and horses.

OBJECTIVE: To study in vitro (1) the dose-response relationships between proteoglycan metabolism in normal and corticosteroid-treated articular cartilage; (2) long-term proteoglycan metabolism after treatment of articular cartilage with corticosteroids; and (3) the effect of corticosteroids on proteoglycan metabolism in articular cartilage treated with monocyte-conditioned medium (MCM). STUDY DESIGN: Equine and canine articular cartilage explants were treated with corticosteroids and MCM. Proteoglycan synthesis and degradation were measured by radioactive labeling in short-term culture, and the long-term effect of corticosteroid treatment on proteoglycan metabolism was studied in normal explants. ANIMALS: Two young cross-breed horses and 3 young Labrador retrievers. METHODS: Equine articular cartilage explants were incubated in medium containing methylprednisolone sodium succinate (MPS) at 0, .001, .01, .1, 1, and 10 mg/mL (final concentration) for 1 day and then in fresh medium without MPS. Proteoglycan synthesis was measured by incorporation of sodium [35S]sulfate at 1, 3, 7, 10, and 13 days after initial treatment with MPS. Proteoglycan release was measured from separate explants prelabeled with sodium [35S]sulfate and treated similarly. Equine articular cartilage explants were treated with equine MCM simultaneously with, and 24 hours before MPS, at 0, 0.01, 0.1, 1, or 5 mg/mL for 72 hours. Proteoglycan synthesis and degradation in these explants was compared. Proteoglycan synthesis and degradation were measured similarly in canine articular cartilage explants treated simultaneously with canine MCM and MPS at 0, 0.001, 0.01, 0.1, 1 and 10 mg/mL for 72 hours. Equine articular cartilage explants treated with 0, 0.01, 0.1, 1, and 5 mg/mL of MPS for 72 hours were evaluated histologically. RESULTS: Proteoglycan synthesis in normal equine articular cartilage was severely depressed by 10 mg/mL MPS for 24 hours, and proteoglycan synthesis failed to recover after 13 days of culture in medium without MPS. Cartilage treated with 5 mg/mL MPS had pyknotic chondrocyte nuclei and empty lacunae. Concentrations of 1 and 0.1 mg/mL MPS depressed proteoglycan synthesis in normal equine cartilage explants. For these 2 concentrations, proteoglycan synthesis recovered 2 days after MPS removal and increased significantly (P < .05) 7 days after treatment with MPS compared with controls without MPS. Concentrations of 0.001 and 0.01 mg/mL MPS did not significantly affect proteoglycan synthesis in normal equine cartilage explants. Cumulative proteoglycan loss over 13 days in culture from normal equine explants treated for 24 hours with different concentrations of MPS was not significantly different between treatment groups at any time point. MCM significantly depressed proteoglycan synthesis in both canine and equine articular cartilage explants and significantly increased proteoglycan release. These effects were prevented in the canine explants by simultaneous treatment with MPS at 1 and 0.1 mg/mL, and proteoglycan release induced by MCM in equine articular cartilage was inhibited by 1 mg/mL MPS. CONCLUSIONS: Concentrations of 1.0 and 0.1 mg/mL MPS alleviated articular cartilage degradation in MCM-treated articular cartilage in vitro. These concentrations of MPS in contact with normal cartilage explants for 24 hours are unlikely to be detrimental in the long term to proteoglycan synthesis. The response of articular cartilage to MPS was affected by treatment with MCM so that results of experiments with normal articular cartilage explants may not reflect results obtained with abnormal cartilage. CLINICAL RELEVANCE: It may be possible to find an intraarticular concentration of corticosteroid that protects articular cartilage against cytokine-induced matrix degradation yet not have prolonged or permanent detrimental effects on chondrocyte matrix synthesis.

Effect of polysulfated glycosaminoglycan on DNA content and proteoglycan metabolism in normal and osteoarthritic canine articular cartilage explants.

OBJECTIVE: To study the effect of polysulfated glycosaminoglycan (PSGAG) on proteoglycan metabolism and DNA content of control and osteoarthritic (OA) cartilage. STUDY DESIGN: An in vitro study comparing the effects of PSGAG on articular cartilage explants from canine stifle joints with and without chronic OA after transection of the left cranial cruciate ligament. SAMPLE POPULATION: Five large cross-breed dogs. METHODS: Cartilage explants (6 to 13 per treatment group) from the medial side of the femoral trochlea and medial femoral condyle from both stifles of each dog were incubated in a defined medium containing 0, 0.05, 0.5, or 5 mg/mL of PSGAG. After 72 hours in culture, explants were pulsed for 6 hours with sodium [35S]sulfate. Aminophenylmercuric acetate (APMA) was used to activate endogenous neutral matrix metalloproteinases (MMPs) and induce proteoglycan degradation in the radiolabeled explants. DNA content and radioactivity were measured in papain-digested explants, and radioactivity was measured in the medium by liquid scintillation counting. Proteoglycan synthesis and degradation were calculated. Cartilage was examined histologically for signs of OA. A mixed model analysis of variance and linear contrasts were used to test for significant (P < .05) effects of OA and treatment with PSGAG. RESULTS: Transection of the cranial cruciate ligament produced OA in operated joints. DNA content and proteoglycan synthesis of OA cartilage were significantly lower than in cartilage from control joints. For both DNA content and proteoglycan synthesis, significant interactions occurred between the concentration of PSGAG and whether the articular cartilage was from OA or control joints. The two lower concentrations of PSGAG (0.05 and 0.5 mg/mL) predominantly increased DNA content in OA cartilage (7 and 18%, respectively, compared with 0 mg/mL PSGAG) while the highest concentration (5 mg/mL) predominantly increased DNA content in control cartilage (30% compared with 0 mg/mL PSGAG). PSGAG at .05 mg/mL predominantly decreased proteoglycan synthesis in OA cartilage (19% reduction compared with 0 mg/mL PSGAG) while PSGAG at .5 and 5 mg/mL predominantly decreased proteoglycan synthesis in control cartilage (17 and 55% reduction, respectively, compared with 0 mg/mL PSGAG). Following activation of MMPs, PSGAG caused a dose-dependent decrease in degradation of radiolabeled proteoglycan in both OA and control cartilage. CONCLUSIONS: OA cartilage was responsive to treatment with PSGAG at 100-fold lower concentration than control cartilage. When treated with PSGAG, articular cartilage explants maintained or increased DNA content at the expense of proteoglycan synthesis. Following MMP activation, proteoglycan degradation was inhibited in OA and control explants in a dose-dependent manner. CLINICAL RELEVANCE: If the results of this study extend to in vivo use, treatment with PSGAG may modify the progression of OA in articular cartilage by maintaining chondrocyte viability or stimulating chondrocyte division as well as protecting against extracellular matrix degradation.

Comparison of the trotting gaits of Labrador Retrievers and Greyhounds.

OBJECTIVE: To compare the trotting gaits of Labrador Retrievers and Greyhounds to determine whether differences in locomotion are attributable to differences in their manner of moving or to body size and shape differences between these 2 breeds. ANIMALS: 8 healthy 5-month-old Greyhounds and 5 healthy Labrador Retrievers between 6 and 18 months old. PROCEDURE: A series of 4 force platforms was used to record independent ground reaction forces on the forelimbs and hind limbs during trotting. Values of stride parameters were compared between breeds before and after normalization for size differences. Standard values of absolute and normalized stride period and stride length were determined from linear regressions of these parameters on relative (normalized) velocity. Forces were normalized to body weight and compared at the same relative velocity. RESULTS: Greyhounds used fewer, longer strides than the Labrador Retrievers to travel at the same absolute speed. After normalization for body size differences, most measurable differences between breeds were eliminated. Subtle differences that did persist related to proportion of the stride that the forefoot was in contact with the ground, timing of initial hind foot contact relative to initial forefoot contact, and distribution of vertical force between the forelimbs and hind limbs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that apparent differences in the trotting gait between Labrador Retrievers and Greyhounds are mainly attributable to differences in size, and that dogs of these 2 breeds move in a dynamically similar manner at the trot.

Acceleration and balance in trotting dogs.

During quadrupedal trotting, diagonal pairs of limbs are set down in unison and exert forces on the ground simultaneously. Ground-reaction forces on individual limbs of trotting dogs were measured separately using a series of four force platforms. Vertical and fore-aft impulses were determined for each limb from the force/time recordings. When mean fore-aft acceleration of the body was zero in a given trotting step (steady state), the fraction of vertical impulse on the forelimb was equal to the fraction of body weight supported by the forelimbs during standing (approximately 60 %). When dogs accelerated or decelerated during a trotting step, the vertical impulse was redistributed to the hindlimb or forelimb, respectively. This redistribution of the vertical impulse is due to a moment exerted about the pitch axis of the body by fore-aft accelerating and decelerating forces. Vertical forces exerted by the forelimb and hindlimb resist this pitching moment, providing stability during fore-aft acceleration and deceleration.

Prognostic factors affecting survival of 507 horses with joint disease: (1983 to 1990).

Between July 1, 1983 and December 31, 1990, risk factors were determined for all horses with joint disease presented to a referral center, of being discharged, of ever becoming sound, or of being alive at 3 mo follow-up. Logistic multiple-regression models were done separately for foals (< or = 4 mo), yearlings (> 4-24 mo) and racing or nonracing adult horses (> 24 mo). The breakdown in this study was 53 foals, 87 yearlings, 141 nonracing adults, and 226 racing adults. Thirty-one foals (58%), 68 yearlings (78%), 119 non-racing adults (84%), and 213 racing adults (94%) were discharged. Foals with a less severe lameness, duration of illness of > 1 d, and infectious arthritis had increased odds of discharge. At follow-up, 12 of 18 (67%) were alive, 10 (56%) of which were sound. Yearlings with osteochondrosis had higher odds of discharge; at follow-up, 38 of 49 (78%) were alive, 32 (65%) of which were sound. For non-racing adults, horses with less severe lameness, without a miscellaneous diagnosis, or intended for pleasure use had increased odds of discharge. At follow-up, 55 of 78 (70%) were alive and 33 of 58 (57%) with soundness data became sound. Risk factors for higher odds of being alive at follow-up were carpal lameness, arthroscopic surgery, a prognosis other than poor, became sound, above-median hospitalization costs, and duration of follow-up. The 161 racing adults (76% of discharges), with follow-up, were more likely to have had osteoarthritis, higher hospital costs, hospitalization > 1 d, and arthroscopy. Sixty-four (60%) of these became sound; the odds increased if the horse was not severely lame at admission or was hospitalized for > 1 d. Risk factors and prognosis differed by age-use group among horses seen at our hospital.

Site-specific variation in femoral head cartilage composition in dogs at high and low risk for development of osteoarthritis: insights into cartilage degeneration.

OBJECTIVE: To determine which characteristics of cartilage lesion pathology are detected in dogs at high risk to develop osteoarthritis prior to diagnosis by standard radiographs or macroscopic cartilage abnormality on necropsy. METHODS: Fourteen disease-free dogs were assigned to risk groups based on hip distraction index. For seven dogs, three dimensional images of hip joints from computed tomography were available. At necropsy, ligamentum capitis femoris volumes were measured and articular cartilage was harvested and analyzed for percent water, swelling, glycosaminoglycan, and fibronectin. Comparisons were made with nine dogs with macroscopic cartilage lesions (OA group). RESULTS: Ligament volumes were greater in the high distraction index group (P=0.000). Water content was elevated in the lesion area in both low and high risk dogs (P=0.000); no additional increase was noted in the high risk group. Glycosaminoglycan content was slightly elevated in the surrounding area in both groups (P< 0.02) but loss was noted histologically in the lesion area of the high risk group. Fibronectin was increased in the lesion area and in the high risk group (P=0.000). The magnitude of this increase was greatest in the lesion area (P=0.000) in explants. Computed tomography indicated dorsal acetabular rim impingement on the lesion area in high risk dogs. CONCLUSIONS: Water content and swelling suggest matrix structure is weaker at the site of lesion predilection in all dogs regardless of risk status. Computed tomography imaging is consistent with site specific initiation of lesions by mechanical factors.

Biomechanical comparison of two plating techniques for fixation of acetabular osteotomies in dogs.

OBJECTIVE: To compare the failure properties of a 5-hole, 2.7-mm curved acetabular plate (AP) to a 5-hole, 3.5-mm reconstruction plate (RP) when applied to acetabular osteotomies. STUDY DESIGN: Cadaver study. ANIMALS OR SAMPLE POPULATION: Pelves of 8 mature, large-breed dogs. METHODS: A 5-hole, 2.7-mm AP and a 5-hole, 3.5-mm RP were contoured and applied to the dorsal acetabulum of each pelvis. A central acetabular fracture was simulated after plate application by a transverse osteotomy with a fine saw. Each acetabulum was loaded in a weight-bearing direction. A load-deformation curve was produced for each construct, and biomechanical properties of the AP and RP were compared with the Student's paired t-test. A P value of < .05 was considered significant. RESULTS: For the AP and RP composite respectively, the mean +/- SD maximum load to failure was 2,721 +/- 632 N and 2,488 +/- 800 N, the stiffness was 4.8 +/- 1.8 N/m and 5.3 +/- 1.9 N/m, and the energy absorbed was 15.1 +/- 5.2 Nm and 16.3 +/- 8.3 Nm. None of these differences was statistically significant. CONCLUSIONS: Both fixation techniques provided comparable strength, stiffness, and energy absorbed under the loading conditions of this study. CLINICAL RELEVANCE: Because of the relative ease of application, the 2.7-mm curved AP may be the practical choice for acetabular fracture repair in large dogs.

An outcrossed canine pedigree for linkage analysis of hip dysplasia.

Canine hip dysplasia (CHD) is a prevalent, debilitating, polygenic disease characterized by hip subluxation and laxity which results in osteoarthritis. We are developing an informative pedigree for linkage analysis of CHD. The seven greyhound founders had excellent hip conformation with high dorsolateral subluxation scores (percentage of femoral head covered by the dorsal acetabulum in a weight-bearing position) of 66 +/- 4% (mean +/- SD averaged over both hips) and low hip distraction (laxity) indices of 0.14 +/- 0.08. Nine greyhounds bred on site had radiographic evidence of ossification in the capital femoral chondroepiphysis at 7.7 +/- 0.9 days of age. At 8 months of age they had a mean distraction index of 0.24 +/- 0.08 and dorsolateral subluxation score of 76 +/- 1%. Of the four dysplastic Labrador retriever founders, three had mean age at onset of capital femoral chondroeplphyseal ossification of 20 +/- 7 days of age n = 3) and a mean distraction index of 0.46 +/- 0.1 accompanied by hip osteoarthritis. Thirty-four F1s had mean onset of capital femoral ossification (10.7 +/- 4.0 days of age) and mean dorsolateral subluxation scores (61 +/- 12%) similar to the greyhound founders, but distraction indices (0.42 +/- 0.2) more similar to the Labrador retriever founders. One F1 had CHD radiographically but none of 20 F1s had osteoarthritis at necropsy at 10 months of age. These data suggested that maximum passive laxity (as measured by the distraction index) and normal osseous conformation (as indicated by a high dorsolateral subluxation score) were both dominant traits and were controlled by separate quantitative trait loci (QTL). Forty-three back-crosses between F1s with the highest hip laxity and greyhound founders had mean onset of capital femoral ossification at 9.9 +/- 2.6 days of age. Of 10 dogs in the backcross generation that have reached 8 months of age, 2 had palpable subluxation without marked CHD radiographically. The mean distraction index of these dogs was 0.36 +/- 0.16 and the dorsolateral subluxation score was 65 +/- 5%. Although dogs in the backcross generation that were three-quarter greyhound had a broad range of hip laxity, a protective effect of the greyhound QTLs for good osseous conformation has mitigated thus far against subluxation and CHD.

Comparison of measurements of dorsolateral subluxation of the femoral head and maximal passive laxity for evaluation of the coxofemoral joint in dogs.

OBJECTIVE: To determine whether dorsolateral subluxation (DLS) of the femoral head reflects osseous conformation of the coxofemoral (hip) joint and represents a property distinct from maximum passive laxity of the hip joint in dogs. ANIMALS: 14 Labrador Retrievers, 16 Greyhounds, 58 Greyhound-Labrador Retriever mixed-breed dogs, and 1 Rottweiler. PROCEDURES: DLS of the femoral head (DLS score) and passive laxity of the hip joint (distraction index) were determined radiographically in 3 groups of dogs: not treated (167 joints of 84 dogs); before and after injecting 2 ml of hyaluronan into 25 hip joints of 13 dogs; and before and after unilateral triple pelvic osteotomy in 5 dogs. Results of the 2 methods were compared for each group. RESULTS: In untreated dogs, the correlation coefficient (r) of DLS score versus distraction index was -0.73 and -0.69 for 84 left and 83 right hip joints, respectively. Mean coefficient of determination (r2) for both hips was 0.5. Mean DLS score did not differ before and after intra-articular injection of hyaluronan into either hip joint, whereas mean distraction index increased significantly after intra-articular injection. Unilateral triple pelvic osteotomy resulted in a significant increase in DLS score, compared with values obtained before surgery. However, distraction index before and after surgery did not differ significantly. CONCLUSIONS AND CLINICAL RELEVANCE: The DLS test assesses the congruity of the acetabulum and the femoral head in a canine hip joint and thus represents a characteristic distinct from maximum passive laxity. The DLS score and the distraction index evaluate different components of hip joint stability.

Dorsolateral subluxation of hip joints in dogs measured in a weight-bearing position with radiography and computed tomography.

OBJECTIVE: develop a radiographic procedure to measure dorsolateral subluxation (DLS) of the femoral head in canine coxofemoral (hip) joints in a weight-bearing position. STUDY DESIGN: DLS measured on a radiographic projection was compared with DLS measured on computed tomography (CT) images of hip joints in a weight-bearing position. ANIMALS: A total of 24 dogs of varying ages were examined including Labrador retrievers, greyhounds, and Labrador-greyhound crossbreeds. METHODS: Anesthetized dogs were placed in sternal recumbency in a kneeling position in a foam rubber mold. The stifles were flexed and adducted with the femora perpendicular to, and in contact with, the table. To test for DLS, dogs were imaged in this weight-bearing position (DLS test) with routine radiography and CT. For each hip, the DLS score was determined by measuring the percentage of the femoral head medial to the lateralmost point of the cranial acetabular rim on the dorsoventral radiographic projection and the lateralmost point of the central, dorsal acetabular rim on the CT image. Higher DLS scores indicated better coverage of the femoral head by the acetabulum. DLS scores were compared with the distraction index (DI) by grouping joints according to their probability of developing osteoarthritis (OA) as predicted by the DI. RESULTS: The DLS score in the new position ranged from 29% to 71% for radiography and 15% to 59% for CT. Joints classified as OA unsusceptible had a mean score of 64% +/- 1.5% for radiography and 55% +/- 0.8% for CT (n = 10); hip joints having a high probability of developing OA had a score of 39% +/- 2.6% for radiography and 26% +/- 1.9% for CT (n = 8). When the DLS test was repeated on the same dogs at a different time, the intraclass correlation coefficient for the DLS score on the radiographs was 0.85 (left hip) and 0.89 (right hip). There was a strong correlation (r = .89 for both hips) between the DLS score measured on the weight-bearing radiograph and the CT image. A strong correlation also was observed between the DLS score and the DI (r = -.87). The DLS scores for OA unsusceptible joints and joints with a high probability of developing OA were significantly different (P < .05). CONCLUSIONS AND CLINICAL RELEVANCE: The DLS test can be performed with CT or routine radiography to measure variable amounts of DLS in weight-bearing hip joints oriented similarly to those of a standing dog. After additional long-term follow-up studies evaluating the development of OA and breed effects are performed, the DLS method may prove useful in studies of normal and abnormal hip joint development related to canine hip dysplasia.

Acute synovitis and intra-articular methylprednisolone acetate in ponies.

OBJECTIVE: To determine how acute synovitis, with and without intra-articular methylprednisolone acetate (MPA), affect synthesis of proteoglycan, total protein, and collagen in articular cartilage and total protein synthesis in synovial membrane. DESIGN: Synovitis was induced in 10 ponies by the injection of 0.5 ng lipopolysaccharide (LPS) into the left radiocarpal and midcarpal joints every 2 days for a total of four treatments. Synovitis was documented by clinical examination and synovial fluid analyses. Two days before euthanasia, MPA (0.1 mg/kg) was injected with the last dose of LPS into both the left and right radiocarpal and midcarpal joints of five of these ponies. Proteoglycan synthesis in articular cartilage explants from these joints was measured by incorporation of sodium [35S]sulfate. The size of the proteoglycan monomers and their aggregation with hyaluronan was assessed by size-exclusion chromatography. Protein synthesis in articular cartilage was measured by incorporation of [3H]proline and collagen synthesis by conversion of [3H]proline into [3H]hydroxyproline. Protein synthesis was measured in synovial membrane explants by incorporation of [35S]methionine. RESULTS: Ponies developed carpal effusion and mild lameness accompanied by increased total nucleated cell count and total solids in synovial fluid in response to the LPS injections. Moderate to severe synovial membrane proliferation and inflammation were observed histopathologically in joints injected with LPS but no consistent light-microscopical changes were observed in the articular cartilage from these joints. Intra-articular MPA alone was associated with decreased proteoglycan synthesis and increased protein and collagen synthesis in the cartilage explants. Total protein synthesis by synovial membrane was also increased by MPA alone. In contrast, no differences in protein or proteoglycan synthesis were observed in explants from the joints with synovitis, with or without intra-articular MPA. Treatment with MPA, LPS, and LPS/MPA did not alter proteoglycan aggregate size, but LPS-induced synovitis resulted in an increase in the second largest population of monomers. MPA increased the synthesis of small proteoglycan monomers. CONCLUSION: Based on the methods used, acute synovitis prevented changes induced by intra-articular MPA alone. Results suggested that the effect of intra-articular MPA on joint metabolism was different between inflamed and normal joints. Experimental studies must consider the effect of inflammation, as well as the potential to introduce in vitro culture artifacts when investigating the effect of intra-articular corticosteroids on chondrocyte function.

Effect of synovitis and corticosteroids on transcription of cartilage matrix proteins.

OBJECTIVE: To determine whether steady-state levels of type-II procollagen, aggrecan core protein, or fibronectin mRNA in articular chondrocytes are altered by synovitis or administration of methylprednisolone acetate (MPA). SAMPLE POPULATION: Articular cartilage specimens collected from 10 ponies, 2.5 to 3.5 years old and 200 to 300 kg. PROCEDURE: 4 experimental groups were compared, using the cartilage specimens: control, MPA-treated, lipopolysaccharide-induced synovitis, and lipopolysaccharide-induced synovitis with MPA treatment. RNA was isolated from articular cartilage and compared by northern blot analysis, using equine-specific cDNA probes. RESULTS: Synovitis increased steady-state levels of type-II procollagen mRNA fivefold and of aggrecan mRNA twofold. Administration of a single intra-articular injection of MPA (0.1 mg/kg of body weight) decreased type-II procollagen transcripts in normal cartilage sixfold, without significant effect on aggrecan or total fibronectin mRNA values. MPA treatment of inflamed joints decreased type-II procollagen and aggrecan mRNA to levels that were not significantly different from those in untreated control specimens. CONCLUSIONS: Articular chondrocytes increase type-II procollagen and aggrecan synthesis in response to synovitis. MPA alters chondrocyte function in normal and inflamed cartilage, suggesting that potential changes in cartilage matrix protein synthesis should be considered when evaluating the therapeutic value of intra-articular administration of corticosteroids. CLINICAL RELEVANCE: Knowledge of how synovitis and corticosteroids (independently and in combination) affect synthesis of cartilage matrix proteins is relevant to understanding pathogenesis of traumatic osteoarthritis and improving therapeutic strategies.