Evaluation of the repair process of cartilage defects of the equine third carpal bone with and without subchondral bone perforation

To determine the effect of subchondral bone drilling (forage) on the cartilage repair process after injury has occurred, a cartilage defect (1 cm in diameter) was created on the radial facet of the proximal surface of each third carpal bone in 6 adult horses. In one of the third carpal bones (right or left thoracic limb) of each horse, a 1-cm cartilage defect was created, and 5 holes (1 mm in diameter and 10 mm deep) were drilled through the subchondral bone into the cancellous bone. In the other thoracic limb, an identical defect was created, but not drilled. Analyses of cell numbers and types in the synovia and the mucin precipitate quality were done before, at 1 week after, and 3 weeks after surgical manipulation was done and showed no significant difference between the joint environment of drilled carpi and those of nondrilled carpi. At 21 weeks after surgical manipulation was done, each joint was examined radiographically, macroscopically, and microscopically to compare the condition of the joints and the state of repair of the cartilage in each defect. The amount of surface of the defect covered by the dense fibrous and fibrocartilagenous repair tissue and the thickness of the repair tissue were significantly greater (P less than 0.05 and P less than 0.01, respectively) in the drilled carpal bones. In addition, the attachment of the repair tissue to underlying chondro-osseous tissue was better in the drilled carpal bones. Fibrocartilage was resurfacing the drilled defects, whereas only fibrous tissue was present in the nondrilled defects.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of subchondral drilling on repair of partial-thickness cartilage defects of third carpal bones in horses

Arthrotomies of middle carpal joints were done on 13 horses, and a 1-cm partial thickness, round defect was made on the radial facet of both third carpal bones. In one joint, 1-mm diameter 1-cm deep holes were drilled within the defect, and one joint was used as a control. horses were assigned to 2 groups--group 1 (n = 6 horses), 5 drill holes; group 2 (n = 7 horses), 11 drill holes. At 1 and 3 weeks after surgery, differences between joints in synovial fluid total protein values, WBC counts, or results of mucin precipitate tests were not significant (P = 0.005). Physically and radiographically, horses were the same during the 12 initial weeks they were housed in stalls and the 9 weeks they were kept in paddocks. Twenty-one weeks after surgery, horses were euthanatized. Joints with drill holes had a significantly greater area (P less than 0.05) of healthy fibrocartilage new tissue: group 1-33 to 68% new tissue, compared with 0 to 23% new tissue in controls; and group 2-22 to 64% new tissue, compared with 0 to 37% new tissue in controls. Differences between healing of defects with drill holes in groups 1 and 2 were not significant. Thickness of new tissue over drill holes was 33 to 61% of thickness of cartilage adjacent to the defect, and thickness of tissue between drill holes was 11 to 43% (group 1) and 8 to 79% (group 2) of the thickness of cartilage adjacent to the defect.(ABSTRACT TRUNCATED AT 250 WORDS)     

Arthroscopic mosaic arthroplasty in the equine third carpal bone

OBJECTIVE--To investigate survival and function of autogenous heterotopic osteochondral grafts in a site where injuries are common. STUDY DESIGN--Three osteochondral grafts were harvested arthroscopically from the femoropatellar joint and transplanted to the third carpal bone (C(3)). Nine months later, histologic, histomorphometric, and biochemical comparisons were made between the transplanted grafts in C(3) and tissue adjacent to the recipient site, the opposing radial carpal bone (C(r)), the donor site in the femoropatellar joint, and the sham-operated contralateral C(3). ANIMALS--One mixed-breed pony and 5 Standardbred horses aged 3 to 8 years old. METHODS--Using instruments modified for equine use, four 4.5-mm-diameter osteochondral grafts were harvested arthroscopically from the distal aspect of the lateral trochlea of the right femur and inserted into the radial facet of the right third carpal bone. The fourth graft was kept as a donor-site control sample. Three months later, regular exercise was started and at 6 months, repeat arthroscopy was conducted to evaluate healing. The horses were euthanatized 9 months after transplantation, and comparisons were made between the grafts, opposing radial carpal bone, and contralateral third carpal bone. The assessment criteria included paravital staining, a modified Mankin scoring system, and biochemical analyses for collagen type, total collagen content, and sulfated glycosaminoglycan concentration. RESULTS--All horses were sound 21 days' postoperatively. At 6 months, all 18 grafts were intact but somewhat soft and opaque compared with surrounding carpal cartilage. Nine months' postoperatively, the bony portions of the grafts were well integrated with the recipient sites, but 6 grafts had histologic evidence of cartilage degeneration. From biochemical analysis of grafts, there was little or no new repair tissue invading the experimental sites, but sulfated glycosaminoglycan (proteoglycan) loss from the transplanted cartilage was marked. CONCLUSIONS--Heterotopic transfer of osteochondral grafts from the distal aspect of the lateral femoral trochlea to the third carpal bone is feasible with minor modifications of human mosaic arthroplasty instruments. The bony portion of the osteochondral grafts was quickly remodeled to provide subchondral support to the transplanted articular cartilage. The loss of proteoglycan from the transplanted cartilage indicates that the grafts might have been injured during harvesting or insertion, or, more likely, did not remodel to meet the demands of a new biomechanical environment. CLINICAL RELEVANCE--These findings suggest that arthroscopic resurfacing of focal osteoarticular defects will not be successful in the long term unless donor and recipient sites can be matched with respect to cartilage thickness, biochemical constituents, and physical properties. Mosaic arthroplasty may be indicated in selected cases in which no other options exist to create a confluent cartilage-covered surface.     

Retrospective study of subchondral sclerosis and lucency in the third carpal bone of Standardbred trotters

The aim was to investigate radiographic findings of subchondral sclerosis and subchondral lucency in the dorsoproximal-dorsodistal (DPr-DDi) projection of the third carpal bone (C3) in relation to clinical appearance and to prognosis for racing. In a retrospective study, case records of 89 Standardbred trotters diagnosed with traumatic carpitis confirmed with intra-articular anaesthesia were examined. Records included data on degree of lameness at presentation and after flexion tests and a radiographic examination of the carpus, including a DPr-DDi projection of the C3. Subchondral lucency was found significantly to influence the degree of lameness at presentation and the time to start but did not significantly affect the chance of racing within 30 months post examination. In the present material no significant relationship between degree of sclerosis and lameness or prognosis for racing within 30 months was found, but the low number of C3 with severe sclerosis limited conclusions about that group.     

Ultrastructural immunolocalisation of bone sialoprotein in the osteocartilagenous interface of the equine third carpal bone

REASONS FOR PERFORMING STUDY: One of the most common causes of lameness in racehorses is osteoarthritis (OA). Pathogenesis is not clear and pathological processes of the different joint tissues interact in often progressive events. The interface between cartilage and newly synthesised bone has been shown to be particularly enriched in bone sialoprotein (BSP), a cell-binding matrix protein. OBJECTIVES: To establish whether changes in the concentration of BSP may serve as a marker for early biochemical changes of the subchondral bone. METHODS: Articular cartilage, cartilage/bone interface and subchondral bone of the proximal third carpal bone from 3 Standardbred trotters were analysed ultrastructurally for the presence of BSP in normal and degenerative areas. RESULTS: A marked increase of BSP in the cartilage/bone interface with degenerative changes of the bone and cartilage compared to the morphologically intact cartilage/bone interface was noted, but levels of the protein were distinctly lower in the distal bone. CONCLUSIONS: The results indicate that BSP has the potential to be used as a marker for changes in bone metabolism in the subchondral bone. POTENTIAL RELEVANCE: Tools to monitor early biochemical changes within the connective tissues of the joint in vivo are essential in studies of the pathogenesis of OA. These could be used to monitor and understand such changes in relation to load, exercise, training programmes, inflammation and the development of OA.     

Ultrastructural immunolocalization of cartilage oligomeric matrix protein (COMP) in the articular cartilage on the equine third carpal bone in trained and untrained horses

The present study was designed to delineate the presence of COMP at the ultrastructural level comparing concentrations between two areas of articular cartilage from the equine third carpal bone, subjected to different loading, from trained and untrained horses. We also analyzed the fibril thickness of collagen type II in the same compartments and zones. Samples were collected from high load-bearing areas of the dorsal radial facet (intermittent high load) and an area of the palmar condyle (low constant load) in five non-trained and three trained young racehorses. The data show that COMP is much less abundant in the matrix in intermittent high loaded areas of articular cartilage from trained horses as compared to the untrained horses (p = 0.036). On the other hand, the untrained horses often displayed a higher immunolabeling in loaded areas compared to unloaded areas, indicating that an adequate dynamic load promotes COMP synthesis and/or retention, while an excessive load may have an opposite effect. The collagen fibril diameter showed marked variation between individuals. The present study indicates that dynamic in vivo compression at high load and frequency lowers matrix content of COMP in the articular cartilage of the third carpal bone. It also indicates that the collagen network is influenced by mechanical load following by strenuous exercise.     

The effect of intra-articular methylprednisolone acetate and exercise on equine carpal subchondral and cancellous bone microhardness

Dorsal carpal osteochondral injury is a major cause of lameness in horses undergoing high intensity training. Intra-articular corticosteroid treatment is used commonly to manage exercise-associated articular pain, but its use remains highly controversial in the equine athlete. This project, therefore, aimed to compare the mechanical properties of intra-articular MPA and diluent-treated middle carpal subchondral and cancellous bone in horses undergoing a short-term treadmill exercise programme. It was hypothesised that subchondral and cancellous bone mechanical properties are influenced by intra-articular administration of methylprednisolone acetate (MPA). Eight 2-year-old female horses had MPA or diluent administered into contralateral middle carpal joints at 14 day intervals, for a total of 4 treatments per horse. Horses underwent a standard treadmill exercise protocol until euthanasia (Day 70). Standard sites were located on the dorsal aspect of third, radial and intermediate carpal bones. Osteochondral samples from each test site were divided into subchondral bone and cancellous bone portions. These were dried, resin-embedded and gold-coated. Microhardness measurements were obtained at each test site. No significant effect of intra-articular treatment was detected. At each site, cancellous bone trabecular struts had an 18-19% higher microhardness value than the overlying subchondral bone. These findings indicate that intra-articular administration of MPA at this dose has no effect on subchondral or cancellous bone adaptation to short-term exercise and, therefore, on the propensity of carpal bones to injury. Further investigation into the calcified cartilage layer, effect of different corticosteroid preparations and diffusion of medication are required.     

Validation of magnetic resonance imaging for measurement of equine articular cartilage and subchondral bone thickness

OBJECTIVE: To validate use of magnetic resonance images (MRIs) for measurement of equine articular cartilage and subchondral bone thickness by comparison with measurements in histologic specimens. SAMPLE POPULATION: 32 cadaveric carpal joints from 16 horses. PROCEDURE: Magnetic resonance imaging was performed by use of 3-dimensional fast spoiled gradient echo (SPGR) and T2* 3-dimensional fast gradient echo (GRE) pulse sequences with and without fat saturation. Standard sites on the medial and lateral facets of the intermediate, radial, and third carpal bones were used for subchondral bone and articular cartilage thickness measurements. Digital image analysis software was used for MRI measurements 10 mm from the dorsal extent and perpendicular to the articular surface. Histomorphometric measurements of hyaline, calcified cartilage, and subchondral bone thickness were obtained at selected sites. Comparisons between histomorphometric and MRI measurements and between magnetic resonance pulse sequences were evaluated. RESULTS: There were significant correlations between GRE and SPGR and SPGR and histologic measurements of articular cartilage, with no significant difference between measurements and good agreement. When calcified cartilage was excluded from the histologic measurement, MRI measurements were significantly greater than histologic measurements. For subchondral bone thickness, there was significant correlation between GRE and SPGR but GRE was significantly greater than SPGR measurements. Histomorphometric and MRI measurements were strongly correlated and not significantly different. CONCLUSIONS AND CLINICAL RELEVANCE: Magnetic resonance imaging provides a good representation of cartilage and subchondral bone thickness, supporting its use in the study and clinical diagnosis of osteochondral structure and alteration.     

Evaluation of subchondral bone mineral density associated with articular cartilage structure and integrity in healthy equine joints with different functional demands

OBJECTIVE: To determine and correlate subchondral bone mineral density and overlying cartilage structure and tensile integrity in mature healthy equine stifle (low magnitude loading) and metacarpophalangeal (high magnitude loading) joints. ANIMALS: 8 healthy horses, 2 to 3 years of age. PROCEDURE: Osteochondral samples were acquired from the medial femoral condyle (FC) and medial trochlear ridge (TR) of the stifle joint and from the dorsal (MC3D) and palmar (MC3P) aspects of the distal medial third metacarpal condyles of the metacarpophalangeal joint. Articular cartilage surface fibrillation (evaluated via India ink staining) and tensile biomechanical properties were determined. The volumetric bone mineral density (vBMD) of the underlying subchondral plate was assessed via dual-energy x-ray absorptiometry. RESULTS: Cartilage staining (fibrillation), tensile moduli, tensile strength, and vBMD were greater in the MC3D and MC3P locations, compared with the FC and TR locations, whereas tensile strain at failure was less in MC3D and MC3P locations than FC and TR locations. Cartilage tensile moduli correlated positively with vBMD, whereas cartilage staining and tensile strain at failure correlated negatively with vBMD. CONCLUSIONS AND CLINICAL RELEVANCE: In areas of high joint loading, the subchondral bone had high vBMD and the articular cartilage surface layer had high tensile stiffness but signs of structural wear (fibrillation and low failure strain). The site-dependent variations and relationships in this study support the concept that articular cartilage and subchondral bone normally adapt to physiologic loading in a coordinated way.     

Radiographic and scintigraphic evaluation of the third carpal bone of control horses and horses with carpal lameness

We compared the radiographic and scintigraphic findings in the third carpal bone of horses performing different work disciplines and investigated their relationship with lameness. Horses had undergone carpal radiography including acquisition of a dorsoproximal-dorsodistal oblique (DPr-DDiO) image of the distal row of carpal bones and/or scintigraphic examination of the carpi. Cause of lameness, breed, age, and work discipline were recorded. Increased opacity in the third carpal bone was graded, ratio of radiopharmaceutical uptake calculated objectively, and increased radiopharmaceutical uptake graded subjectively. Relationships between radiographic, scintigraphic, and clinical findings were assessed statistically. Increased opacity in the third carpal bone (P = 0.003) and ratio of radiopharmaceutical uptake (P = 0.015) were associated with the work discipline. Increased opacity in the third carpal bone was associated with both increased radiopharmaceutical uptake grade (P = 0.002; rs = 0.59) and ratio of radiopharmaceutical uptake (P = 0.013; rs = 0.46). Increased radiopharmaceutical uptake and increased opacity in the third carpal bone were not always observed concurrently. Lameness related to the middle carpal joint was associated with increased opacity (P < 0.001), ratio of radiopharmaceutical uptake (P = 0.037), and increased radiopharmaceutical uptake grade (P < 0.001). Radiographic and scintigraphic abnormalities were observed in horses performing all disciplines, indicating that high-speed exercise may not be the only factor determining the development of osseous disease in the third carpal bone. Both increased opacity and increased radiopharmaceutical uptake were more likely to be seen in horses with lameness related to the middle carpal joint than in horses with other sources of pain.     

Intra-osseous pressure of the equine third metatarsal bone

Intra-osseous pressure was measured in the third metatarsal bone of 8 pony foals at the age of 1, 3, 6 and 26 weeks of age, and on one occasion in the third metatarsal bone of 5 ponies aged 1-5 years; the animals were under general anesthesia and in lateral recumbency. Saphenous venous pressure and lateral superficial plantar metatarsal arterial pressure were also monitored throughout the procedure. There were no statistically significant changes in the epiphyseal, diaphyseal, venous or arterial pressures with increasing age of the foals. There was no statistically significant difference between the mean epiphyseal and mean diaphyseal pressures in the foals. Individually however, the diaphyseal pressure exceeded the epiphyseal pressure in 32 of the 36 measurements made in the foals. Saphenous vein occlusion and intravenous adrenalin administration showed that both venous and arterial pressures influence intra-osseous pressure. Intra-osseous injection of heparinized saline on one side of the growth plate produced an immediate massive increase in intra-osseous pressure on that side of the growth plate, but in almost all cases a concomitant rise in intraosseous pressure on the other side of the physis did not occur. Simulated effusion of the metatarsophalangeal joint was associated with an increase in intraosseous pressure in the adjacent metatarsal epiphysis, and flexion of the joint with an increase in intraosseous pressure in both the epiphysis and the metaphysis.     

The distribution of cartilage oligomeric matrix protein (COMP) in equine carpal articular cartilage and its variation with exercise and cartilage deterioration

Based on previous studies where tendons receiving the most load have been shown to have the highest levels of cartilage oligomeric matrix protein (COMP), we hypothesized that COMP distribution in articular cartilage may be influenced by mechanical loading. This investigation aimed (a) to describe the pattern of COMP immunoreactivity in middle carpal joint cartilage of two-year-old Thoroughbred horses; (b) to determine topographical variations; (c) to compare high (group 1) and low (group 2) intensity training and (d) to describe COMP immunoreactivity at sites with early osteoarthritis.Group 1 (n =6) underwent a 19 week high-intensity treadmill training programme and group 2 (n =6) were given daily walking until euthanasia. Dorsal and palmar sites on radial and third carpal articular surfaces were prepared. Immunohistochemistry was performed with polyclonal rabbit anti-equine COMP antiserum using a biotin-streptavidin/peroxidase method. Results showed: (a) intracellular immunoreactivity was present in all cartilage zones, but the distribution of COMP staining within the matrix varied between cartilage zones; (b) differences in distribution between sites were not observed, but total COMP levels in exercised horses (n =2) did vary between sites with dorsal sites containing less COMP than palmar sites on the radial, intermediate and third carpal lateral facet; (c) group 1 cartilage showed marked interterritorial distribution in the deep layer compared to group 2 where staining was more generalized throughout the matrix and (d) fibrillated cartilage showed increased local immunoreactivity in the matrix. These findings demonstrate zonal variations in equine COMP distribution which may be influenced by loading.     

Clinical effects of exercise on subchondral bone of carpal and metacarpophalangeal joints in horses

OBJECTIVE: To determine effects of treadmill exercise on subchondral bone of carpal and metacarpophalangeal joints of 2-year-old horses. ANIMALS: 12 healthy 2-year-old horses. PROCEDURE: Horses were randomly assigned to the control (n = 6) or exercised (6) groups. Horses in the exercised group ran on a high-speed treadmill 5 d/wk for 6 months. Horses in the control group were hand walked for the same amount of time. Results of clinical, radiographic, nuclear scintigraphic, and computed tomographic examinations, and serum and synovial concentrations of biochemical markers of bone metabolism were compared between groups. RESULTS: Exercised horses were significantly lamer at the end of the study than control horses. Radionuclide uptake in the metacarpal condyles, but not in the carpal joints, was greater in exercised horses, compared with control horses. Exercised horses also had a higher subchondral bone density in the metacarpal condyles than control horses, but such differences were not detected in the carpal bones. CONCLUSIONS AND CLINICAL RELEVANCE: None of the diagnostic techniques evaluated was sufficiently sensitive to detect all osteochondral damage. Computed tomography and computed tomographic osteoabsorptiometry were superior to conventional radiography for detecting small osteochondral fragments. Nuclear scintigraphy was a sensitive indicator of subchondral bone change but lacked specificity for describing lesions and discerning normal bone remodeling from damage. Newer techniques such as computed tomography may help clinicians better diagnose early and subtle joint lesions in horses prior to development of gross joint damage.     

Mechanical and morphometric analysis of the third carpal bone of Thoroughbreds

The third carpal bone (C3) was collected from both forelimbs of 27 Thoroughbreds. On the basis of age, training, and history, specimens were assigned to 1 of 5 groups: yearling, untrained horses (group 1, n = 4); 2- to 3-year-old, untrained horses (group 2, n = 7); trained 2-year-old horses (group 3, n = 6); trained 3-year-old horses (group 4, n = 6); and 3-year-old, trained horses with carpal pathologic features (group 5, n = 4). A transverse section of subchondral bone 5-mm thick was cut in a precise fashion 10 mm below the proximal articular surface of all specimens. After high-detail radiography was done, indentation testing was performed on the proximal surface of the section at points 5 mm apart. The stiffness of the subchondral cancellous bone was determined from the slope of the load vs displacement curve. Topographic plots of stiffness measurements were compared with radiographs of each specimen. Point determinations were averaged to derive measures for the radial and intermediate facets, and for regions 5, 10, 15, and 20 mm from the dorsal margin of C3. Area fraction (1-p; p = porosity) was measured for the radial and intermediate facets, using an automated image analysis system. Significant (P less than 0.05) increases in stiffness and area fraction were found in the C3 from trained horses (groups 3 to 5), compared with untrained horses (groups 1 to 2). Stiffness and area fraction of the radial facet of pathologic C3 were significantly higher than the same variables measured in C3 from any other group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison between magnetic resonance imaging,computed tomography,and arthrography to identify artificially induced cartilage defects of the equine carpal joints

While articular cartilage changes are considered to be one of the initial events in the pathological cascade leading to osteoarthritis, these changes remain difficult to detect using conventional diagnostic imaging modalities such as plain radiography. The aim of this prospective, experimental, methods comparison study was to compare the sensitivity of magnetic resonance imaging (MRI), magnetic resonance arthrography, computed tomography (CT), and CT arthrography in the detection of artificially induced articular cartilage defects in the equine carpal joints. Defects were created in the antebrachiocarpal and middle carpal joint using curettage by a board‐certified equine surgeon. Normal articular cartilage thickness varied from a maximum of 1.22 mm at the level of the distal aspect of the radius to a minimum of 0.17 mm in the proximal articular surface of the third carpal bone. Regarding cartilaginous defect measurements the remaining cartilaginous bed range from a maximum of 0.776 mm in the partial thickness defects, and 0 mm (defect reaches the subchondral bone) when total thickness defect were made. Computed tomography and magnetic resonance imaging were performed followed by CT arthrography and magnetic resonance arthrography after antebrachiocarpal and middle carpal intraarticular contrast administration. All images were reviewed by two board‐certified veterinary radiologists, both of whom were blinded to the location, presence of, and thickness of the cartilage defects. A total number of 72 lesions in nine limbs were created. Mean sensitivity for localizing cartilage defects varied between imaging modalities with CT arthrography showing the best sensitivity (69.9%), followed by magnetic resonance arthrography (53.5%), MRI (33.3%), and CT (18.1%) respectively. The addition of contrast arthrography in both magnetic resonance and CT improved the rate of cartilage lesion detection although no statistical significance was found. Computed tomographic arthrography displayed the best sensitivity for detecting articular cartilage defects in the equine antebrachiocarpal and middle‐carpal joints, compared to magnetic resonance arthrography, MRI, and CT.     

Results of treatment of subchondral bone cysts in the medial condyle of the equine femur with an autogenous cancellous bone graft

The results of surgical treatment of 10 subchondral bone cysts, all located in the medial femoral condyle, are presented. A cancellous bone graft was used in nine cases and a two component acrylic bone cement was used in the case of an extremely large cyst. Eight out of the 10 cases made a satisfactory clinical recovery.     

Effects of intramuscular polysulfated glycosaminoglycan on chemical and physical defects in equine articular cartilage.

The effect of intramuscular polysulfated glycosaminoglycan (PSG) on repair of cartilage injury was evaluated in eight horses. In each horse, one middle carpal joint had both a partial-thickness and a full-thickness articular cartilage defect created. In the contralateral middle carpal joint, chemical articular cartilage injury was created by intra-articular injection of 50 mg sodium monoiodoacetate (MIA). Horses were divided into two groups for treatment. Group 1 horses (control) received an intramuscular injection of normal saline every four days for a total of seven injections starting seven days after cartilage injury. Group 2 horses received 500 mg of PSG intramuscularly every four days for seven treatments starting seven days after cartilage injury. Horses were maintained for 12 weeks. Horses were evaluated clinically, and their middle carpal joints were evaluated radiographically and arthroscopically at the end of the study. Joint tissues were also collected and examined microscopically. The only significant difference between groups was slightly greater matrix staining intensity for glycosaminoglycans in the radiate articular cartilage layer in MIA injected and PSG treated joints. Partial-thickness defects had not healed and the predominant repair tissue in full-thickness defects was fibrous tissue. It was concluded that using this joint injury model, 500 mg PSG administered intramuscularly had no effect on the healing of articular cartilage lesions, and minimal chondroprotective effect from chemically induced articular cartilage degeneration.     

The quantitative assessment of photodensity of the third carpal bone in the horse

AIM: To determine whether variation in x-ray-beam angle significantly affected photodensity of the third carpal bone (C3) in the horse using the tangential radiographic view, and indirectly determine whether radioabsorptiometry (RA) could be used to assess differences in bone mineral density (BMD) of C3 between exercised and non-exercised horses. METHODS: The left distal carpal row was isolated post-mortem from 2-year-old Thoroughbred horses that had been either exercised (n=7) for up to 14 weeks using a standard training regimen for racehorses, or not exercised (n=7). The effect of variation in x-ray-beam angle on photodensity was determined on these isolated carpal bones in vitro. Image analysis was used to assess photodensity (compared to a known thickness of aluminium) of four regions of interest (ROI) in C3 and one ROI in the fourth carpal bone (C4) of 14 isolated distal rows of carpal bones of the horse. The isolated carpal bones were placed fl at on a x-ray cassette and radiographed at 90 degrees (i.e. with the x-ray beam perpendicular to the cassette). The x-ray-beam angle was varied in the dorsal sagittal plane by 5 degrees increments to a total of 15 degrees from 90 degrees and from a base angle of 60 degrees (the x-ray-beam angle at which the tangential view is taken in clinical cases). RESULTS: Variation in beam angle of <10 degrees from 90 degrees significantly affected photodensity, and photodensity was significantly affected when the angle was varied <5 degrees from 60 degrees . When taken at an x-ray-beam angle of 60 degrees , the abaxial aspect of the radial facet of C3 had a consistently higher photodensity than the rest of C3 and C4. The photodensity of the third and fourth carpal bones was higher in exercised horses than in non-exercised horses. CONCLUSION: As variation in x-ray-beam angle significantly affected photodensity, RA using the tangential view is not considered clinically applicable for assessing BMD of C3 and the accuracy of subjective assessment of BMD of C3 using the tangential view in horses is questionable.