OBJECTIVE To compare results obtained with a handheld reference point indentation instrument for bone material strength index (BMSi) measurements in the equine third metacarpal bone for various testing conditions.
SAMPLE 24 third metacarpal bones.
PROCEDURES Third metacarpal bones from both forelimbs of 12 horses were obtained. The dorsal surface of each bone was divided into 6 testing regions. In vivo and ex vivo measurements of BMSi were obtained through the skin and on exposed bone, respectively, to determine effects of each testing condition. Difference plots were used to assess agreement between BMSi obtained for various conditions. Linear regression analysis was used to assess effects of age, sex, and body weight on BMSi. A mixed-model ANOVA was used to assess effects of age, sex, limb, bone region, and testing condition on BMSi values.
RESULTS Indentation measurements were performed on standing sedated and recumbent anesthetized horses and on cadaveric bone. Regional differences in BMSi values were detected in adult horses. A significant linear relationship (r 2 = 0.71) was found between body weight and BMSi values. There was no difference between in vivo and ex vivo BMSi values. A small constant bias was detected between BMSi obtained through the skin, compared with values obtained directly on bone.
CONCLUSIONS AND CLINICAL RELEVANCE Reference point indentation can be used for in vivo assessment of the resistance of bone tissue to microfracture in horses. Testing through the skin should account for a small constant bias, compared with results for testing directly on exposed bone.
OBJECTIVE To investigate the role of the major equine acute phase protein serum amyloid A (SAA) in inflammation of equine intraarticular tissues.
SAMPLE Articular chondrocytes and fibroblast-like synoviocytes (FLSs) from 8 horses (4 horses/cell type).
PROCEDURES Chondrocytes and FLSs were stimulated in vitro for various periods up to 48 hours with cytokines (recombinant interleukin [IL]-1β, IL-6, tumor necrosis factor-α, or a combination of all 3 [IIT]) or with recombinant SAA. Gene expression of SAA, IL-6, matrix metalloproteinases (MMP)-1 and −3, and cartilage-derived retinoic acid-sensitive protein were assessed by quantitative real-time PCR assay; SAA protein was evaluated by immunoturbidimetry and denaturing isoelectric focusing and western blotting.
RESULTS All cytokine stimulation protocols increased expression of SAA mRNA and resulted in detectable SAA protein production in chondrocytes and FLSs. Isoforms of SAA in lysed chondrocytes and their culture medium corresponded to those previously detected in synovial fluid from horses with joint disease. When exposed to SAA, chondrocytes and FLSs had increased expression of IL-6, SAA, and MMP3, and chondrocytes had increased expression of MMP-1. Chondrocytes had decreased expression of cartilage-derived retinoic acid-sensitive protein.
CONCLUSIONS AND CLINICAL RELEVANCE Upregulation of SAA in chondrocytes and FLSs stimulated with proinflammatory cytokines and the proinflammatory effects of SAA suggested that SAA may be involved in key aspects of pathogenesis of the joint inflammation in horses.
OBJECTIVE To compare regional proportions and spatial distributions of volumetric bone mineral density (BMDv) of the palmar aspect of the distal epiphysis of the third metacarpal bone (McIII) in limbs with or without a condylar fracture from Thoroughbred racehorses.
SAMPLE McIIIs from cadavers of Thoroughbred racehorses with (n = 6 bones) and without (8) a condylar fracture.
PROCEDURES BMDv and spatial distributions of BMDv in peripheral quantitative CT images of the distal epiphysis of McIIIs were quantitatively assessed with spatial analysis software. Relative proportions of voxels within 9 threshold categories of BMDv and spatial statistics for BMDv distribution were compared between fractured and nonfractured limbs.
RESULTS No significant differences in BMDv characteristics were identified between fractured and nonfractured limbs, although fractured limbs had a lower proportion of voxels in the BMDv thresholds 700 to < 800 mg/cm3 and 800 to < 900 mg/cm3 but a higher proportion of voxels in the BMDv threshold 1,000 to < 1,100 mg/cm3 for the central condylar region of the medial condyle. Results of spatial analysis reflected the response of bone to race training rather than differences between fractured and nonfractured limbs. In both limb groups, uniform clusters of low BMDv with areas of high BMDv were identified.
CONCLUSIONS AND CLINICAL RELEVANCE BMDv characteristics of the distal epiphysis of McIII reflected training load, and fracture characteristics were subtle. Serial imaging techniques in conjunction with detailed training data are required to elucidate the onset of the pathological response to load in horses.
OBJECTIVE To determine by use of an in vitro model the potential for translocating sufficient numbers of bacteria into a joint during arthrocentesis through cellulitic tissue to cause sepsis.
SAMPLE Culture media containing 4 concentrations of Staphylococcus aureus and needles of 3 sizes.
PROCEDURES Needles (22, 20, and 19 gauge) were inserted through Mueller-Hinton agar that contained known concentrations of S aureus (103,104,105, and 106 CFUs/mL). After a needle exited through the medium, any agar plug within the needle bore was ejected into a sterile syringe and the contaminated portion of the needle was harvested. Sterile saline (0.9% NaCl) solution was used to emulsify the agar plug and wash the contaminated portion of the needle. The resulting solution was cultured to determine the number of bacterial CFUs that could be deposited into a joint during arthrocentesis through contaminated tissue.
RESULTS Needle gauge and bacterial concentration were both associated with the number of bacterial CFUs deposited after insertion through contaminated agar. Although all needle sizes were capable of bacterial translocation sufficient to cause septic arthritis, ORs for 20- and 22-gauge needles translocating > 33 CFUs of S aureus were significantly higher than the OR for a 19-gauge needle. The ORs for 20- or 22-gauge needles translocating > 33 CFUs of S aureus (the minimum population of S aureus known to cause joint sepsis) were 0.22.
CONCLUSIONS AND CLINICAL RELEVANCE Results for this in vitro model indicated that caution should be used when performing arthrocentesis through cellulitic tissue.
OBJECTIVE To compare serum concentrations of biomarkers of cartilage and bone metabolism between racehorses with a carpal or metacarpophalangeal or metatarsophalangeal (ie, fetlock) joint injury and matched uninjured control horses, determine changes in biomarker concentrations following joint injury, and establish the biomarkers’ diagnostic test performance.
ANIMALS 50 Thoroughbred racehorses with a carpal or fetlock joint injury and 50 matched uninjured horses (control horses).
PROCEDURES Serum concentrations of 2 cartilage synthesis biomarkers (carboxy-terminal propeptide of type II collagen [CPII] and chondroitin sulfate epitope 846 [CS846]), 2 cartilage degradation biomarkers (neoepitope generated by collagenase cleavage of type II collagen [C2C] and cross-linked carboxy-terminal telopeptide fragments of type II collagen [CTX-II]), and serum activity of a bone formation marker (bone-specific alkaline phosphatase [BAP]) were measured around the time of injury diagnosis and monthly thereafter for as long as possible.
RESULTS Injured horses as a group and horses specifically with fetlock joint injuries had significantly lower serum CPII concentrations and significantly higher serum BAP activities than matched control horses. Concentrations of CTX-II were decreased between 2 and 4 months following joint injury. Measurement of CPII concentration at baseline could distinguish between injured horses and control horses with a sensitivity of 82% and specificity of 50%.
CONCLUSIONS AND CLINICAL RELEVANCE Although significant differences in specific biomarker concentrations between horses with carpal and fetlock joint injuries and matched control horses were identified, there was no convincing evidence of the suitability of these biomarkers as diagnostic or prognostic tools in a clinical setting.
OBJECTIVE To quantify concentrations of cartilage oligomeric matrix protein (COMP) and fibromodulin in synovial fluid from the tarsocrural joints (TCJs) of horses with osteochondritis dissecans (OCD) of the distal intermediate ridge of the tibia and determine whether concentrations would change following arthroscopic removal of osteochondral fragments.
ANIMALS 115 client-owned horses with OCD of the TCJ and 29 control horses euthanized for unrelated reasons.
PROCEDURES COMP and fibromodulin concentrations were measured in synovial fluid from the TCJs of the affected horses before and after osteochondral fragments were removed arthroscopically and in synovial fluid from the TCJs of the control horses after euthanasia. Synovial biopsy specimens from the TCJs of affected and control horses were examined histologically for evidence of inflammation.
RESULTS Synovial fluid COMP and fibromodulin concentrations prior to surgery in horses with OCD were not significantly different from concentrations in control horses. Fibromodulin, but not COMP, concentration in horses with OCD was significantly decreased after surgery, compared with the concentration before surgery. Fibromodulin concentration was significantly correlated with joint effusion score but not with lameness score or results of a flexion test and was correlated with histologic score for number of synoviocytes on the surface of the synovium but not with score for degree of infiltration of inflammatory cells in the synovium. Synovial fluid COMP concentration was not significantly correlated with clinical or histologic findings.
CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that fibromodulin, but not COMP, could potentially be a biomarker of joint inflammation in horses with OCD of the TCJ.
OBJECTIVE To test ex vivo mechanical properties of 4 allograft fixation techniques for cranial cruciate ligament (CCL) replacement.
SAMPLE 30 stifle joints from canine cadavers.
PROCEDURES CCL-deficient stifle joints repaired by 1 of 4 techniques (n = 6/group) and CCL-intact stifle joints (control group; 6) were mechanically tested. Three repair techniques involved a patella-patella ligament segment (PPL) allograft: a tibial and femoral interference screw (PPL-2S), a femoral interference screw and the patella seated in a tapering bone tunnel in the tibia (PPL-1S), or addition of a suture and a bone anchor to the PPL-1S (PPL-SL). The fourth technique involved a deep digital flexor tendon (DDFT) allograft secured with transverse femoral fixation and stabilized with a tibial interference screw and 2 spiked washers on the tibia (DDFT-TF). The tibia was axially loaded at a joint angle of 135°. Loads to induce 3, 5, and 10 mm of femoral-tibia translation; stiffness; and load at ultimate failure with the corresponding displacement were calculated. Group means were compared with a multivariate ANOVA.
RESULTS Mean ± SD load for the intact (control) CCL was 520.0 ± 51.3 N and did not differ significantly from the load needed to induce 3 mm of femoral-tibial translation for fixation techniques PPL-SL (422.4 ± 46.3 N) and DDFT-TF (654.2 ± 117.7 N). Results for the DDFT-TF were similar to those of the intact CCL for all outcome measures.
CONCLUSIONS AND CLINICAL RELEVANCE The DDFT-TF yielded mechanical properties similar to those of intact CCLs and may be a viable technique to test in vivo.
OBJECTIVE To evaluate associations of measures assessed by radiography, 2-D CT, and 3-D CT of the hip joints of immature dogs with osteoarthritis in the same joints at maturity.
ANIMALS 46 hound-type dogs from a colony predisposed to osteoarthritis.
PROCEDURES Images of hip joints (1/dog) were obtained at 16, 32, and 104 weeks of age. Radiographic measures included Norberg angle, distraction index, and osteoarthritis score. Two-dimensional CT measures included acetabular index, percentage of femoral head coverage, and center edge, horizontal toit externe, acetabular anteversion, and ventral, dorsal, and horizontal acetabular sector angles. Three-dimensional CT measures were femoral head and neck volume, femoral neck angle, and femoral head and neck radius. Differences among measures at 16 and 32 weeks in dogs with different osteoarthritis scores at later time points, relationships among variables at each time point, and relationships of single and combined measures with the presence of osteoarthritis at 104 weeks were evaluated.
RESULTS The 16- and 32-week distraction index, center edge angle, dorsal acetabular sector angle, horizontal acetabular sector angle, percentage of femoral head coverage, acetabular index, and Norberg angle and the 32-week femoral neck angle varied significantly with osteoarthritis severity at 104 weeks. Presence of osteoarthritis in mature dogs was most strongly associated with 16-week combined measures of distraction index and center edge angle and 32-week combined measures of dorsal acetabular sector angle and Norberg angle.
CONCLUSIONS AND CLINICAL RELEVANCE Changes in hip joint morphology associated with radiographic signs of osteoarthritis were detectable as early as 16 weeks of age and varied with osteoarthritis severity in adult dogs. The use of combined hip joint measures may improve early identification of dogs predisposed to hip joint osteoarthritis.
OBJECTIVE To evaluate whether a low-dosage regimen of prednisolone induces bone loss and whether administration of alendronate sodium prevents glucocorticoid-induced osteopenia in dogs by measuring trabecular bone mineral density (BMD) with quantitative CT.
ANIMALS 8 healthy Beagles.
PROCEDURES In 4 dogs, prednisolone was administered PO at a dosage of 2 mg/kg once daily for 2 weeks, 1 mg/kg once daily for 4 weeks, and 0.5 mg/kg once daily for 3 weeks. In the other 4 dogs, alendronate sodium (2 mg/kg, PO, q 24 h) was whether administered for 9 weeks in addition to the same dosage of prednisolone used in the prednisolone-treated dogs. Before (day 0 [baseline]) and 21, 42, 63, and 150 days after the start of treatment, BMD of the lumbar vertebrae was measured by quantitative CT.
RESULTS BMD in the prednisolone treatment group decreased to 84.7% of the baseline value on day 42, increased to 87.9% on day 63, and recovered to 91.6% on day 150. In the prednisolone-alendronate treatment group, BMD decreased to 91% of the baseline value on day 21, increased to 93.8% on day 63, and then recovered to 96.7% on day 150. Bone mineral density in the prednisolone treatment group was generally lower, albeit not significantly, than that of the prednisolone-alendronate treatment group on each examination day.
CONCLUSIONS AND CLINICAL RELEVANCE BMD temporarily decreased after low-dosage prednisolone administration; however, it gradually improved during tapering of the prednisolone dosage. These results have suggested that a low dosage of prednisolone can be used with little concern for development of osteopenia in dogs.
Objective—To determine whether stromal cell-derived factor-1 (SDF-1) concentrations in serum, plasma, and synovial fluid differed among untrained, race-trained, and osteochondral-injured Thoroughbred racehorses.
Animals—22 racehorses without osteochondral injury and 37 racehorses with osteochondral injury.
Procedures—Horses without osteochondral injury were examined before and after 5 to 6 months of race training. Horses with osteochondral injury were undergoing arthroscopic surgery for removal of osteochondral fragments from carpal or metacarpophalangeal or metatarsophalangeal joints (fetlock joints). Serum, plasma, and fetlock or carpal synovial fluid samples were obtained and analyzed for SDF-1 concentration by use of an ELISA.
Results—In horses with fetlock or carpal joint injury, mean synovial fluid SDF-1 concentrations were significantly higher, serum SDF-1 concentrations were significantly lower, and synovial fluid-to-serum SDF-1 ratios were significantly higher than in untrained and trained horses. Synovial fluid SDF-1 concentrations were not significantly different between trained and untrained horses. Plasma SDF-1 concentrations were not different among the 3 groups. Results obtained with serum, compared with synovial fluid and plasma, had better sensitivity for differentiating between osteochondral-injured horses and uninjured horses. In horses with fetlock joint osteochondral injury, serum SDF-1 concentrations were correlated with radiographic and arthroscopic inflammation scores, but not arthroscopic cartilage scores.
Conclusions and Clinical Relevance—Results suggested that serum SDF-1 concentrations were more sensitive than plasma and synovial fluid concentrations for detection of osteochondral injury in the fetlock or carpal joint of racehorses. Analysis of serum and synovial SDF-1 concentrations in horses with experimentally induced joint injury may help define the onset and progression of post-traumatic osteoarthritis and aid in the evaluation of anti-inflammatory treatments.