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.     

Healing of full-thickness cartilage compared with full-thickness cartilage and subchondral bone defects in the equine third carpal bone.

The effect of lesion depth on the quality of third carpal bone cartilage repair was examined. A 1-cm diameter articular defect penetrating the calcified cartilage in one limb and the subchondral bone plate in the opposite limb was created in the radial facet of the third carpal bones. Clinical and xeroradiographic examinations were performed every 4 weeks until 4 months (3 horses) and 6 months (3 horses) after surgery. The synovial membrane, non-opposing articular surfaces and articular defects were examined grossly, histologically and histochemically. Grossly, deeper defects contained thicker, whiter tissue, but both joints contained generalised degenerative changes. Defects extending through calcified cartilage were filled deeply by fibrocartilage and superficially by fibrous connective tissue. Defects extending through subchondral bone were consistently filled with hyaline-like cartilage in the depths of the lesion, fibrocartilage in the intermediate layer and fibrous connective tissue superficially. The results indicate that subchondral bone is the source of hyaline-like cartilage repair tissue and suggest that quality of healing of cartilage defects may be improved by penetrating the subchondral bone plate. It also appears that the synovitis associated with the procedure must be controlled before the procedure can be advocated for treatment of clinical cases.     

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)     

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.     

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)     

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.     

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.     

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.     

Technique for arthroscopic repair of third carpal bone slab fractures in horses

A technique involving internal fixation guided by arthroscopy was used to repair third carpal slab fractures in 23 horses. The technique allowed good reduction and compression of the fractures, with minimal surgical trauma. Ten of 17 horses with a 6-month or longer follow-up have returned to race successfully.     

Arthroscopic landmarks of the equine carpus

The radiocarpal and middle carpal joints of 4 clinically normal horses and 24 necropsy specimens were examined with an arthroscope to describe the topographic anatomy of these joints. The carpal bones of the radiocarpal joint had congruent articular surfaces in extension, but carpal flexion resulted in a stairstep between the radial and intermediate carpal bones. The variable surface anatomy, as well as the restricted synovial space and the technical limitations of the arthroscope, contributed to the need for more than one arthroscopic approach to the radiocarpal joint to ensure a thorough examination. The middle carpal joint's hinge-like biomechanics resulted in a spacious synovial cavity that was more amenable to arthroscopic examination. Careful planning was necessary to ensure that areas of interest were distant from the arthroscope-insertion site to allow the best perspective possible. The lateral approach to the middle carpal joint provided the best viewing of both facets of the intermediate and third carpal bones as well as the dorsal rim of the radial carpal bone. The medial approach did not allow complete inspection of these areas.     

Pathophysiology and diagnosis of third carpal bone disease in horses: a review

Third carpal bone (C3) disease is a significant cause of lameness in Standardbred and Thoroughbred horses. The bone density of C3 increases as a result of exercise, reducing the compliance of the bone and predisposing it to injury. Currently, the most widely used method of diagnosis is subjective radiography using the tangential view. Radiographically, increases in bone mineral density (BMD) appear as sclerosis but it is not known at what point increases in sclerosis indicate the onset of disease or increased risk of C3 fracture. A quantitative assessment of the BMD of C3 in horses would improve understanding of the changes that occur within this bone and guide athletic management, as it is thought that BMD changes precede articular cartilage damage. Methods of non-invasive bone-mineral analysis used for the detection of osteoporosis in humans include single photon absorptiometry (SPA), dual x-ray absorptiometry (DXA), computed tomography (CT), radioabsorptiometry (RA), quantitative ultrasonography (QU) and magnetic resonance imaging (MRI). To date, DXA and RA are the most commonly used methods of quantitative non-invasive bone-mineral analysis in horses. The cost of equipment and difficulties in performing DXA in live animals preclude the routine use of this technique for diagnostic purposes. RA may become clinically applicable to C3 analysis in horses, but small variations in x-ray beam angle when taking the tangential view significantly affect results, making this technique clinically inapplicable at this time. Currently, methods of quantitative non-invasive bone-mineral analysis of C3 in horses are not suited to clinical application.     

Intra-osseous pressure and pressure pulse gradients along the equine third metatarsal bone

Intra-osseous pressure measurements in the third metatarsal bone of anesthetised young and mature ponies demonstrated that a pressure gradient exists along the bone, the pressure being highest at the level of the nutrient foramen and decreasing distally and proximally from this point. The height of the intra-osseous pressure pulse also decreased toward the bone ends. It is concluded that the sites of measurement of the intra-osseous pressure in clinical and experimental studies should be standardized.     

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.     

Tenoscopic release of the equine carpal canal

OBJECTIVE: To develop a tenoscopic method to divide the carpal flexor retinaculum and decompress the carpal canal. STUDY DESIGN: Cadaver specimen study and prospective trial. ANIMALS: Twelve cadaveric limbs, 4 clinically normal horses, and 2 clinically affected horses. Methods-A tenoscopic approach to the proximolateral aspect of the carpal sheath was used to identify and facilitate endoscopic division of the inner and outer layers of the carpal flexor retinaculum in cadaver limbs. The technique was further evaluated in 4 normal and 2 clinically affected horses. RESULTS: Anatomic dissection, intraoperative observation, necropsy, histologic evaluation, and both short- and long-term clinical follow-up indicate that a tenoscopic approach that divides the inner layer of the carpal retinaculum can successfully decompress the equine carpal canal. No iatrogenic damage to surrounding structures was evident, division of the retinaculum was adequate and permanent, and clinical morbidity was negligible. Resolution of effusion was evident in both clinical cases of carpal canal syndrome and lameness resolved in the 1 horse in which long-term follow-up was possible. CONCLUSIONS: Tenoscopic release of the carpal flexor retinaculum could provide a minimally invasive method to quickly, safely, and effectively decompress the carpal canal. CLINICAL RELEVANCE: Tensocopic release of the carpal flexor retinaculum is a safe alternative to open division of the retinaculum to decompress the carpal canal in horses with carpal canal syndrome.     

A biomechanical comparison of headless tapered variable pitch and AO cortical bone screws for fixation of a simulated slab fracture in equine third carpal bones

OBJECTIVE: To compare the mechanical shear strengths and stiffnesses obtained from in vitro testing of a simulated complete third carpal bone (C3) frontal plane radial facet slab fracture (osteotomy) stabilized with either a 4/5 Acutrak (AT) compression screw or a 4.5-mm AO cortical bone (AO) screw inserted in lag fashion. Drilling, tapping, and screw insertion torques, forces, and times also were compared between AT and AO implants. STUDY DESIGN: In vitro biomechanical assessment of site preparation, screw insertion, and shear failure test variables of bone screw stabilized simulated C3 slab fracture in paired cadaveric equine carpi. SAMPLE POPULATION: Eight pairs of cadaveric equine C3 without orthopedic abnormalities. METHODS: Standardized simulated C3 slab fractures were repaired with either AO or AT screws (AO/C3 and AT/C3 groups, respectively). Drilling, tapping, and screw insertion torques, forces, and times were measured with a materials testing machine for each screw type. Repaired specimens were tested in axially oriented shear until failure. Paired Students t-tests were used to assess differences between site preparation, screw insertion, and shear testing variables. Significance was set at P <.05. RESULTS: There were no significant differences in bone fragment measurements of the standardized simulated C3 slab fractures created for AO or AT screws. There were no significant differences for mean and maximum drilling torques; however, the tapered AT drill had greater maximum drilling force compared with the 3.2-mm and 4.5-mm AO drill bits. Mean insertion torque and force measured from the self-tapping AT screw were not significantly different compared with the 4.5-mm AO tap. There were no significant differences in maximum screw torque among constructs. Total procedure time was significantly longer for the AT group (5.8 +/- 1.6 minutes) compared with the AO group (2.9 +/- 1.1 minutes; P =.001). AT stabilized specimens had significantly greater mean +/- SD initial shear stiffness (3.64 +/- 1.08 kN/mm) than AO specimens (1.64 +/- 0.73 kN/mm; P =.005). All other shear mechanical testing variables were not statistically different among screw types. CONCLUSION: The 4/5 Acutrak insertion technique was accurate and safe, and the AT screw effectively stabilized simulated equine C3 frontal plane slab fractures. When tested in shear, this screw type was mechanically comparable to the 4.5-mm AO screw; however, AT constructs had greater initial shear stiffness. Initial shear stiffness was likely an indirect measure of interfragmentary compression, and thus may indicate that the AT screw provides a more rigid fixation for frontal plane C3 slab fractures in horses. CLINICAL RELEVANCE: Considering the comparable mechanical behavior, greater initial shear stiffness for AT screw stabilized C3 slab fracture fragments, the ability to accurately insert the screw with the aid of a guide pin, and the potential for less persistent soft tissue irritation with the headless screw design, the 4/5 tapered AT screw is an attractive alternative for repair of C3 slab fractures in horses.