Objective—To determine fibroblast viability, assess
development of apoptosis, and evaluate tissue hypoxia
via histochemical, in-situ hybridization, or immunohistochemical
staining in ruptured and intact cranial
cruciate ligaments (CCLs) of dogs.
Animals—32 dogs with ruptured CCLs, and 8 aged
and 19 young dogs with intact CCLs.
Procedure—Markers of cell viability (lactate dehydrogenase
[LDH]), apoptosis (terminal deoxynucleatidyl
transferase-mediated deoxyuridine triphosphate-nick
end labeling [TUNEL] method), and hypoxia (hypoxiainducible
factor-1α [HIF-1α] monoclonal antibody)
were applied to CCL specimens; positive cells were
assessed objectively (LDH) and subjectively (TUNEL
and HIF-1α) in the main axial tissue component (core)
and synovial intima and subintima (epiligamentous tissue).
Results—Viable fibroblasts were seen in all intact and
ruptured CCLs. More nonviable cells were found in
the core regions of ruptured CCLs and intact CCLs of
young dogs than in the epiligamentous regions.
Number of nonviable cells in the core region of ruptured
CCLs was greater than that in intact CCLs of
young and aged dogs, whereas the number in the
epiligamentous region was similar in all specimens.
The TUNEL and HIF-1αstaining was only found in the
epiligamentous region of ruptured CCLs.
Conclusions and Clinical Relevance—Ruptured
CCLs contained a high number of nonviable cells but
not a great number of apoptotic cells. Repair processes
in the epiligamentous region of the CCL include a
metabolic response to hypoxia, suggesting that
necrosis of ligament fibroblasts and transformation of
surviving cells to a spheroid phenotype may be a
response to hypoxia cause by microinjury or inadequate
blood flow. (Am J Vet Res 2003;64:1010–1016)
Objective—To determine those bones in the distal
aspect of the limbs of Greyhounds with fatigue fractures
that have the greatest left-to-right differences in
bone-mineral density (BMD).
Sample Population—Limbs obtained from 20
Procedure—Dual-energy x-ray absorptiometry (DXA)
of the distal aspect of each limb and isolated bones
from 10 dogs with a fracture of the central tarsal bone
(CTB) of the right pelvic limb was performed. Highresolution
scanning was performed on excised
bones, and BMD measurements of CTB also were
obtained from limbs of dogs without a CTB fracture.
Results—The BMD of the accessory carpal bone and
calcaneus was not significantly different from the
BMD of those bones in the contralateral limb.
Although BMD of the CTB of the entire right pelvic
limb and isolated bones from dogs with a CTB fracture
was lower, compared with values for the entire
left pelvic limb, values for isolated CTB from dogs
without a CTB fracture were not significantly different.
Metacarpal or metatarsal and thoracic or pelvic
limb significantly affected BMD for measurements of
the entire limb and isolated bones. Left-to-right differences
in BMD were greatest for metacarpal 5.
Conclusions and Clinical Relevance—Asymmetric
adaptive remodeling of metacarpal 5 can be detected
by DXA. The potentially confounding effects of CTB
fracture and unknown racing history made it difficult
to interpret BMD changes in the CTB of these specimens.
Densitometry could be developed as an in vivo
assessment for risk of fractures in racing
Greyhounds. (Am J Vet Res 2000;61:1214–1219)
To evaluate the diagnostic capabilities of a novel helical fan beam CT system used for imaging of horses with a range of clinical distal limb problems.
Medical records were reviewed of horses presented for CT of the distal limb at 2 university-based veterinary hospitals. The following data were recorded: age, sex, breed, presenting complaint, sedation used for imaging, scanning time, procedure time, other diagnostic imaging methods performed, imaging diagnosis, clinical diagnosis, and complications during imaging.
Most horses were Thoroughbreds and Quarter Horses. Procedure times ranged from 15 to 40 minutes, with scanning completed in 15 to 45 seconds for each region of interest. The foot or pastern region was commonly scanned (88/167 [53%] horses), with navicular bone disease diagnosed in 42 of 88 (48%) horses. The fetlock region was also commonly scanned (42/167 [40%] horses), with palmar or plantar osteochondral disease diagnosed in 17 of 42 (40%) horses. Horses were compliant during scanning, and no complications with sedation or damage to the scanner occurred. A specific imaging diagnosis for the lameness was achieved more frequently with CT imaging (166/167 [99%]) than with planar digital radiography (26/58 [45%]).
The helical fan beam CT system could be used safely to scan sedated standing horses from the carpal or tarsal region distally. Subjectively, the machine was easy to operate, allowing CT to be incorporated into lameness investigations. CT imaging was very likely to result in a clinical diagnosis in horses with distal limb lameness.
To evaluate the diagnostic capabilities of a novel helical fan beam CT system used for imaging of horses with clinical problems of the head and neck.
Medical records were reviewed of horses presented for CT of the head or neck at 2 university hospitals. The following data were recorded: age, sex, breed, presenting complaint, sedation used for imaging, scanning time, procedure time, other diagnostic imaging methods performed, imaging diagnosis, clinical diagnosis, and complications during imaging.
Quarter Horses and Warmbloods were the most common breeds, and the most common complaint was nasal discharge. The head (101/120 [84%] horses) was scanned most frequently, and the most common diagnoses were primary dental disease and a space-occupying lesion of the paranasal sinuses. Nuchal bursitis was the most common imaging diagnosis in the neck region. Procedure time ranged from 20 to 45 minutes with a scanning time of 30 to 40 seconds. No complications with horse sedation occurred, and horses tolerated scanning well. An imaging diagnosis was more frequently achieved with CT (109/120 [91%] horses) than with planar digital radiography (23/61 [38%] horses).
The helical fan beam CT system had fast scanning times and could be used safely for routine imaging of the teeth and sinuses in horses. The caudal extent of scanning in the neck region was limited by the shape of the horse’s neck and thorax.
Objective—To compare 2 methods of quantitating chondrocyte viability and to determine chondrocyte response to thermal injury over time.
Sample Population—108 stifle joints from 54 adult rats.
Procedures—Cartilage from the distal aspect of the femur was treated ex vivo with radiofrequency energy at a probe setting that would result in immediate partial-thickness chondrocyte death; untreated sections served as controls. Explants were cultured, and cell viability was compared by use of lactate dehydrogenase (LDH) histochemical staining and calcein AM and ethidium homodimer-1 confocal laser microscopy (CLM) cell viability staining. Terminal deoxynucleotidyl transferase–mediated X-dUTP nick end labeling (TUNEL) was used to detect apoptosis. All labeling studies were performed 0, 1, 3, 7, 14, and 21 days after treatment.
Results—In the treated tissues, a greater percentage of viable cells were found with CLM, compared with LDH staining. This result contrasted that of control tissues in which LDH staining indicated a greater percentage of live cells than CLM. The greatest number of TUNEL-positive chondrocytes was present at day 3, declining at later time intervals.
Conclusions and Clinical Relevance—CLM and LDH histochemistry techniques yield different absolute numbers of live and dead cells, resulting in differing percentages of live or dead cells with each technique. These differences may be related to the enzymes responsible for activation in each technique and the susceptibility of these enzymes to thermal injury. Results of TUNEL indicate that apoptosis contributes to chondrocyte death after thermal injury, with a peak signal identified 3 days after insult.
Objective—To compare expression of tartrate-resistant
acid phosphatase (TRAP) and cathepsin K and
histologic changes in canine cranial cruciate ligaments
(CCLs) and human anterior cruciate ligaments (ACLs).
Study Population—Sections of cruciate ligaments
from 15 dogs with ruptured CCLs, 8 aged dogs with
intact CCLs, 14 human beings with ruptured ACLs,
and 11 aged human beings with intact ACLs.
Procedure—The CCLs and ACLs were evaluated histologically,
and cells containing TRAP and cathepsin K
were identified histochemically and immunohistochemically,
Results—The proportion of ruptured CCLs that contained
TRAP+ cells was significantly higher than the
proportion of intact ACLs that did but similar to proportions
of intact CCLs and ruptured ACLs that did.
The proportion of ruptured CCLs that contained
cathepsin K+ cells was significantly increased, compared
with all other groups. Numbers of TRAP+ and
cathepsin K+ cells were significantly increased in ruptured
CCLs, compared with intact ACLs. The presence
of TRAP+ cells was correlated with inflammatory
changes, which were most prominent in ruptured
Conclusion and Clinical Relevance—Results suggest
that synovial macrophage-like cells that produce
TRAP are an important feature of the inflammation
associated with CCL rupture in dogs. Identification of
TRAP and cathepsin K in intact CCLs and ACLs from
aged dogs suggests that these enzymes have a functional
role in cruciate ligament remodeling and repair.
We hypothesize that recruitment and activation of
TRAP+ macrophage-like cells into the stifle joint synovium
and CCL epiligament are critical features of the
inflammatory arthritis that promotes progressive
degradation and eventual rupture of the CCL in dogs.
(Am J Vet Res 2005;66:2073–2080)
OBJECTIVE To determine whether walking at specific ranges of absolute and relative (V*) velocity would aid efficient capture of gait trial data with low ground reaction force (GRF) variance in a heterogeneous sample of dogs.
ANIMALS 17 clinically normal dogs of various breeds, ages, and sexes.
PROCEDURES Each dog was walked across a force platform at its preferred velocity, with controlled acceleration within 0.5 m/s2. Ranges in V* were created for height at the highest point of the shoulders (withers; WHV*). Variance effects from 8 walking absolute velocity ranges and associated WHV* ranges were examined by means of repeated-measures ANCOVA.
RESULTS The individual dog effect provided the greatest contribution to variance. Narrow velocity ranges typically resulted in capture of a smaller percentage of valid trials and were not consistently associated with lower variance. The WHV* range of 0.33 to 0.46 allowed capture of valid trials efficiently, with no significant effects on peak vertical force and vertical impulse.
CONCLUSIONS AND CLINICAL RELEVANCE Dogs with severe lameness may be unable to trot or may have a decline in mobility with gait trial repetition. Gait analysis involving evaluation of individual dogs at their preferred absolute velocity, such that dogs are evaluated at a similar V*, may facilitate efficient capture of valid trials without significant effects on GRF. Use of individual velocity ranges derived from a WHV* range of 0.33 to 0.46 can account for heterogeneity and appears suitable for use in clinical trials involving dogs at a walking gait.
OBJECTIVE To develop contact time (ConT) and withers height-normalized relative ConT (ConT*) for force platform gait analysis of dogs.
ANIMALS 29 healthy client-owned dogs.
PROCEDURES Height at the most dorsal aspect of the shoulders (withers) was measured with a framing square. Dogs were trotted across a force platform at their preferred velocity with controlled acceleration (± 0.5 m/s2). Ranges of ConT and ConT* centered on the population mean ConT were created. Variance effects on ground reaction forces (GRFs) for 4 thoracic limb and 4 pelvic limb ConT and associated ConT* ranges were examined. Efficiency of trial capture and effects of velocity ranges on GRF variance were determined.
RESULTS Individual dogs had the greatest effect on GRF variance for thoracic and pelvic limbs. Narrow ConT and ConT* ranges had few significant effects on GRFs but were inefficient at capturing trials. The ConT ranges of 0.22 to 0.29 seconds and 0.19 to 0.25 seconds for thoracic and pelvic limbs, respectively, provided the most efficient rates of trial capture with the fewest significant effects on GRFs. Compared with ConT and ConT* ranges, relative velocity ranges had higher efficiency and smaller GRF variance effects.
CONCLUSIONS AND CLINICAL RELEVANCE Dogs of various morphologies have differing limb velocities. Use of ConT as a surrogate for limb velocity may improve GRF data quality. We identified ConT and ConT* ranges associated with low GRF variance. However, relative velocity ranges captured data more efficiently. Efficient capture of data may help avoid worsening of lameness during gait analysis of dogs.
Objective—To determine the effects of 2 doses of recombinant human bone morphogenetic protein-2 in an absorbable collagen sponge (rhBMP-2/ACS) on bone healing in dogs.
Animals—27 adult dogs.
Procedures—Dogs underwent a mid-diaphyseal (1-mm) tibial osteotomy (stabilized with external skeletal fixation) and received an ACS containing 0.28 mg (0.2 mg/mL) or 0.56 mg (0.4 mg/mL) of rhBMP-2 or no treatment (control dogs). All dogs were examined daily; bone healing was assessed via radiography and subjective lameness evaluation every 2 weeks. After euthanasia at 8 weeks, tibiae were evaluated biomechanically and histologically.
Results—Control dogs required antimicrobial treatment for pin-site–related complications more frequently than did rhBMP-2/ACS–treated dogs. At 4 and 6 weeks, weight bearing was greater in dogs treated with rhBMP-2/ACS (0.2 mg/mL) than in control dogs, albeit not significantly. Compared with control treatment, both doses of rhBMP-2/ACS accelerated osteotomy healing at 4, 6, and 8 weeks, and the 0.2 mg/mL dose enhanced healing at 2 weeks; healing at 6 weeks was greater for the lower-dose treatment than for the higher-dose treatment. Histologically, healing at 8 weeks was significantly improved for both rhBMP-2/ACS treatments, compared with control treatment. Among groups, biomechanical variables did not differ, although less osteotomy-site failures occurred in rhBMP-2/ACS–treated groups.
Conclusions and Clinical Relevance—In dogs that underwent tibial osteotomy, rhBMP-2/ACS (0.2 mg/mL) appeared to accelerate bone healing and reduce lameness (compared with control treatment) and apparently augmented bone healing more than rhBMP-2/ACS (0.4 mg/mL). Compared with control dogs, rhBMP-2/ACS–treated dogs required antimicrobial treatments less frequently.
OBJECTIVE To assess the effects of sedation on results of acoustoelastography of the superficial digital flexor tendons (SDFTs) in clinically normal horses.
ANIMALS 27 clinically normal horses.
PROCEDURES For each horse, the pathology index (PI) for the SDFT of each thoracic limb was determined by use of acoustoelastography at 4 locations (5, 10, 15, and 20 cm distal to the accessory carpal bone). Horses were evaluated before and after they were sedated with a combination of detomidine hydrochloride (0.01 mg/kg, IV) and butorphanol tartrate (0.01 mg/kg, IV). A repeated-measures ANOVA was used for statistical analysis.
RESULTS Overall, the PI was lower after sedation than before sedation. In addition, the PI was lower at more distal locations than at more proximal locations. There was not a significant effect of limb (left or right). Differences among individual horses accounted for the largest variance effect.
CONCLUSIONS AND CLINICAL RELEVANCE Sedation with detomidine and butorphanol facilitated acoustoelastography; however, it decreased the SDFT PI in clinically normal horses and should be used consistently in prospective studies. Variance associated with each individual horse in the sample population had the greatest effect on the PI.