Serial sections of bone and soft tissue from the metacarpophalangeal joints of 2 mature and 2 immature horses were evaluated for substance P immunoreactive sensory nerve fibers. Formalin-fixed specimens were sectioned, either nondemineralized or demineralized with formic acid or EDTA. Rabbit antiserum to substance P (SP) was used in the strep. tavidin-biotin-peraxidase complex method for immunolocalization of SP antigen, and staining with 3,3'- diaminobenzidine was used for permanent identification of SP fibers. Abundant sensory nerve fibers were identified in the joint capsule, synovial membrane subintimal layers, collateral ligaments, suspensory ligament and distal sesamoidean ligament attachments to the sesamoid bones, and the periarticular periosteal layers. Sparse SP-immunoreactive nerve fibers were found in subchondral bone plates of the metacarpus, proximal first phalanx, and dorsal articular surface of the sesamoid bones. Most SP fibers were associated with blood vessels in the small cancellous spaces and haversian canals of the subchondral bone. The deeper marrow spaces contained increased numbers of SP sensory fibers; a few appeared in small groups and as several SP-immunoreactive fibers in a larger nerve. Cortical bone contained only a few SP fibers in the haversian canals. Substance P fibers were not identified in the osteocytic lacunae, canaliculi, or the bony lamellae of the haversian systems of the subchondral bone plate, and its extension to the metaphyseal and diaphyseal cortical bone. Equine metacarpophalangeal joint soft tissues have an abundant sensory nerve supply, similar to that of other species. However, the subchondral bone plate also has sparse sensory nerve fibers, which is a unique finding, and may help explain signs of bone pain associated with disease states of the fetlock.

Effect of fluoride was assessed on molars during and after mineralization. Two groups of 7 sheep each were dosed orally with 3.5 mg of fluoride/kg of body weight daily for 4 months (from 5 to 9 months after birth). Sheep of the first group were slaughtered immediately after fluoride administration; those of the second group were slaughtered 4 months later at the age of 13 months. Three control groups of 7 sheep each were slaughtered at 5 months (to determine the state of the teeth at the beginning of fluoride administration), and at 9 and 13 months. During fluoride administration, plasma fluoride concentration rapidly increased to about 0.50 micrograms/ml; after fluoride administration, it stabilized at 0.20 micrograms/ml in treated sheep, whereas controls had concentration of 0.10 micrograms/ml (P < 0.01). Parts of the molars that were in the process of mineralization during fluoride administration (mainly second molars) had thinning enamel, with pits, mainly close to the apex, marked decrease in hardness throughout the layer (< 100 Vickers U, compared with 240 Vickers U), and fluoride accumulation twice as high as that in controls (1,000 to 2,500 mg(kg [dry weight]). Fluoride accumulation was higher in dentine (2,700 to 4,200 mg/kg), but hardness was less affected. On parts of the molars that were already mineralized mostly, the first molar), changes in the appearance of enamel and cementum, decreased hardness (less important than in teeth during mineralization) affecting outer enamel more than inner enamel, high fluoride concentration (4,000 to 5,500 mg(kg [dry weight]) in outer enamel extending over 200 Km were observed. Thus, in sheep, fluoride has a substantial postsecretory effect that may be explained by a slower maturation phase of enamel in this species. Because molar wear is correlated to enamel hardness (dentine at the occlusal surface has low resistance--30 Vickers U), abnormal abrasion of molar teeth that have mineralized before and during fluoride intakes can be observed.

Of 143 Greyhounds necropsied consecutively, 6 (4%) had chondrocalcinosis of the scapulohumeral joint; lesions were identified in 6 additional dogs. Lesions were seen exclusively in the humeral head, mainly in the plateau region. The lesions in the dogs of the initial group were unilateral, but 2 of the 6 additional dogs had bilateral lesions. Focal mineralization of articular cartilage appeared as a white raised nidus, sometimes surrounded by a translucent halo in the opaque cartilage. Circular, small translucent cartilage foci, with or without beginning mineralization, were adjacent to definitive chondrocalcinosis lesions. Chondrocyte necrosis and matrix degradation were considered to antedate appearance of matrical mineral granules; mineralization of the cartilage was considered a secondary process, but not necessarily an epiphenomenon. Scanning electron microscopy indicated that the chondrocalcinosis lesion was composed of deposits of irregularly fused stone material that, in scanning and transmission electron micrographs, was composed of irregular spheroids, 0.05 to 0.5 micrometer in diameter. The spheroids contained poorly formed needle-like crystals of apatite. Sparse transformation of the mineral phase into hydroxyapatite was considered to be attributable to a biological mechanism that inhibited phase transition. Cartilage degeneration and chondrocalcinosis of the plateau region of the humeral head appear to be unique lesions that develop in young Greyhounds. It is possible that these lesions are the result of the biomechanical stress of training and racing.

The normal radiographic anatomy of the equine larynx was determined by use of xeroradiography and dissection. The body and laminae of the thyroid cartilage, the muscular process of the arytenoid cartilages, and the dorsal lamina and arch of the cricoid cartilage had radiographic evidence of mineralization (calcification) and/or ossification in clinically normal horses. There was a significant (P less than 0.01) increase in the degree of mineralization of the thyroid and arytenoid cartilages with advancing age. Horses with diagnosis of arytenoid chondrosis (arytenoid chondral dysplasia, arytenoid chondropathy) by use of endoscopy had radiographic changes that included: enlargement with increased density of the arytenoid cartilage region, abnormal patterns of mineralization (dystrophic mineralization or osseous metaplasia), abnormal contour of the corniculate process(es) and laryngeal masses, sometimes obliterating part or all of the lateral laryngeal ventricles.

Objective-To evaluate the use of CT and MRI for guidance of osteochondral sample collection for histologic detection of early osteoarthritic lesions in centrodistal (distal intertarsal) joints of horses. Sample -Right tarsal joints from the cadavers of 24 Icelandic horses aged 29 to 31 months. Procedures-CT and MRI were used to evaluate the extent of suspected osteoarthritic changes in centrodistal joints, which were graded with a semiquantitative system. The anatomic regions with the highest grade of change were identified, and osteochondral samples were obtained from these regions. Samples were also obtained from the same centrodistal joints at predetermined sites. Histologic examination of all samples was performed, with samples classified as negative or positive for osteoarthritis, and results were compared between sample collection methods. Results-Histologic examination revealed osteoarthritic lesions in 29% (7/24) of centrodistal joints with the predetermined method and in 63% (15/24) with the image-guided method. Significant associations were identified between histologic osteoarthritis detection and the summed image-guided sample collection site image grades, central osteophytes, articular cartilage thickness abnormalities, grade 2 articular mineralization front defects, and grade 2 marginal osteophytes. Conclusions and Clinical Relevance-CT and MRI aided the detection of focal changes suggestive of early-stage osteoarthritis in the centrodistal joints of equine cadavers and may be useful for detection of similar disease in live horses. The first morphological changes of centrodistal joint osteoarthritis were suspected to be in the articular cartilage and the articular mineralization front regions.

Serial sections of bone and soft tissue from the metacarpophalangeal joints of 2 mature and 2 immature horses were evaluated for substance P immunoreactive sensory nerve fibers. Formalin-fixed specimens were sectioned, either nondemineralized or demineralized with formic acid or EDTA. Rabbit antiserum to substance P (SP) was used in the strep. tavidin-biotin-peraxidase complex method for immunolocalization of SP antigen, and staining with 3,3'- diaminobenzidine was used for permanent identification of SP fibers. Abundant sensory nerve fibers were identified in the joint capsule, synovial membrane subintimal layers, collateral ligaments, suspensory ligament and distal sesamoidean ligament attachments to the sesamoid bones, and the periarticular periosteal layers. Sparse SP-immunoreactive nerve fibers were found in subchondral bone plates of the metacarpus, proximal first phalanx, and dorsal articular surface of the sesamoid bones. Most SP fibers were associated with blood vessels in the small cancellous spaces and haversian canals of the subchondral bone. The deeper marrow spaces contained increased numbers of SP sensory fibers; a few appeared in small groups and as several SP-immunoreactive fibers in a larger nerve. Cortical bone contained only a few SP fibers in the haversian canals. Substance P fibers were not identified in the osteocytic lacunae, canaliculi, or the bony lamellae of the haversian systems of the subchondral bone plate, and its extension to the metaphyseal and diaphyseal cortical bone. Equine metacarpophalangeal joint soft tissues have an abundant sensory nerve supply, similar to that of other species. However, the subchondral bone plate also has sparse sensory nerve fibers, which is a unique finding, and may help explain signs of bone pain associated with disease states of the fetlock.

Objective: To determine morphological and mechanical properties of trabecular bone of horses with a bone fragility syndrome (BFS; including silicate-associated osteoporosis). Sample: Cylindrical trabecular bone samples from the distal aspects of cadaveric third metacarpal bones of 39 horses (19 horses with a BFS [BFS bone samples] and 20 horses without a BFS [control bone samples]). Procedures: Bone samples were imaged via micro-CT for determination of bone volume fraction; apparent and mean mineralized bone densities; and trabecular number, thickness, and separation. Bone samples were compressed to failure for determination of apparent elastic modulus and stresses, strains, and strain energy densities for yield, ultimate, and failure loads. Effects of BFS and age of horses on variables were determined. Results: BFS bone samples had 25% lower bone volume fraction, 28% lower apparent density, 18% lower trabecular number and thickness, and 16% greater trabecular separation versus control bone samples. The BFS bone samples had 22% lower apparent modulus and 32% to 33% lower stresses, 10% to 18% lower strains, and 41 % to 52% lower strain energy densities at yield, ultimate, and failure loads, compared with control bone samples. Differences between groups of bone samples were not detected for mean mineral density and trabecular anisotropy. Conclusions and Clinical Relevance: Results suggested that horses with a BFS had osteopenia and compromised trabecular bone function, consistent with bone deformation and pathological fractures that develop in affected horses. Effects of this BFS may be systemic, and bones other than those that are clinically affected had changes in morphological and mechanical properties.

Effect of fluoride was assessed on molars during and after mineralization. Two groups of 7 sheep each were dosed orally with 3.5 mg of fluoride/kg of body weight daily for 4 months (from 5 to 9 months after birth). Sheep of the first group were slaughtered immediately after fluoride administration; those of the second group were slaughtered 4 months later at the age of 13 months. Three control groups of 7 sheep each were slaughtered at 5 months (to determine the state of the teeth at the beginning of fluoride administration), and at 9 and 13 months. During fluoride administration, plasma fluoride concentration rapidly increased to about 0.50 micrograms/ml; after fluoride administration, it stabilized at 0.20 micrograms/ml in treated sheep, whereas controls had concentration of 0.10 micrograms/ml (P < 0.01). Parts of the molars that were in the process of mineralization during fluoride administration (mainly second molars) had thinning enamel, with pits, mainly close to the apex, marked decrease in hardness throughout the layer (< 100 Vickers U, compared with 240 Vickers U), and fluoride accumulation twice as high as that in controls (1,000 to 2,500 mg(kg [dry weight]). Fluoride accumulation was higher in dentine (2,700 to 4,200 mg/kg), but hardness was less affected. On parts of the molars that were already mineralized mostly, the first molar), changes in the appearance of enamel and cementum, decreased hardness (less important than in teeth during mineralization) affecting outer enamel more than inner enamel, high fluoride concentration (4,000 to 5,500 mg(kg [dry weight]) in outer enamel extending over 200 Km were observed. Thus, in sheep, fluoride has a substantial postsecretory effect that may be explained by a slower maturation phase of enamel in this species. Because molar wear is correlated to enamel hardness (dentine at the occlusal surface has low resistance--30 Vickers U), abnormal abrasion of molar teeth that have mineralized before and during fluoride intakes can be observed.