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- Author or Editor: C. Wayne McIlwraith x
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Abstract
Objective—To investigate effects of 1% hyaluronic acid–chondroitin sulfate–N-acetyl glucosamine (HCNAG) on the damage repair response in equine articular cartilage.
Sample—Articular cartilage from 9 clinically normal adult horses.
Procedures—Full-thickness cartilage disks were harvested from the third metacarpal bone. Cartilage was single-impact loaded (SIL) with 0.175 J at 0.7 m/s and cultured in DMEM plus 1 % (vol/vol) HCNAG or fibroblastic growth factor (FGF)-2 (50 ng/mL). Histologic and immunohistochemical techniques were used to identify tissue architecture and apoptotic cells and to immunolocalize type I and II collagen and proliferating nuclear cell antigen (PCNA).
Results—Type II collagen immunoreactivity increased in SIL cartilage, compared with control samples. At days 14 and 28 (day 0 = initiation of culture), control samples had significantly fewer repair cells than did other treatment groups. In control samples and SIL + HCNAG, there was a significant decrease in apoptotic cell number, compared with results for SIL and SIL + FGF-2 samples. At days 14 and 28, there was a significant increase in chondrocytes stained positive for PCNA in the control samples.
Conclusions and Clinical Relevance—1% HCNAG significantly affected apoptotic and repair cell numbers in an SIL damage-repair technique in adult equine articular cartilage. However, HCNAG had no effect on the number of PCNA-positive chondrocytes or on type II collagen immunohistochemical results. The inclusion of 1% HCNAG in lavage solutions administered after arthroscopy may be beneficial to cartilage health by increasing the number of repair cells and decreasing the number of apoptotic cells.
Abstract
Objective—To evaluate the effect of fibrin concentrations on mesenchymal stem cell (MSC) migration out of autologous and commercial fibrin hydrogels.
Sample—Blood and bone marrow from six 2- to 4-year-old horses.
Procedures—Autologous fibrinogen was precipitated from plasma and solubilized into a concentrated solution. Mesenchymal stem cells were resuspended in fibrinogen solutions containing 100%, 75%, 50%, and 25% of the fibrinogen precipitate solution. Fibrin hydrogels were created by mixing the fibrinogen solutions with MSCs and thrombin on tissue culture plates. After incubation for 24 hours in cell culture medium, the MSCs that had migrated onto the tissue culture surface and beyond the boundary of the hydrogels were counted. This procedure was repeated with a commercial fibrin sealant.
Results—Hydrogel-to-surface MSC migration was detected for all fibrin hydrogels. Migration from the 25% autologous hydrogels was 7.3-, 5.2-, and 4.6-fold higher than migration from 100%, 75%, and 50% autologous hydrogels, respectively. The number of migrating cells from 100%, 75%, and 50% autologous hydrogels did not differ significantly. With commercial fibrin sealant, the highest magnitude of migration was from the 25% hydrogels, and it was 26-fold higher than migration from 100% hydrogels. The 75% and 50% hydrogels resulted in migration that was 9.5- and 4.2-fold higher than migration from the 100% hydrogels, respectively.
Conclusions and Clinical Relevance—MSC migration from fibrin hydrogels increased with dilution of the fibrinogen component for both autologous and commercial sources. These data supported the feasibility of using diluted fibrin hydrogels for rapid delivery of MSCs to the surface of damaged tissues.
Abstract
Objective—To validate an equine inertial measurement unit (IMU) system rigidly attached to a hoof against a 3-D optical kinematics system in horses during walking and trotting.
Animals—5 clinically normal horses.
Procedures—5 swing phases of the hooves of the right forelimb and hind limb were collected via both 3-D optical and IMU systems from 5 horses during walking and trotting. Linear and angular positions, velocities, and accelerations were compared between the 2 systems.
Results—Of the 55 variables compared between the 2 systems, 25 had high correlations (r > 0.8) and 18 had moderate correlations (r > 0.5). Root mean squared errors were lowest in the sagittal plane and orientation (1.1 to 4.4 cm over a range of 1.5 to 1.9 m in the cranial-caudal direction and 2.5° to 3.5° over a range of 88° to 110° rotating around the medial-lateral axis). There were more differences between the 2 systems during small changes in motion, such as in the medial-lateral and proximal-distal directions and in the angular measures around the cranial-caudal and proximal-distal axes.
Conclusions and Clinical Relevance—The equine IMU system may be appropriate for rigid attachment to a hoof of a horse and use in examination of linear and angular motion in the sagittal plane of the hoof during the swing phase while walking and trotting. Although promising in many respects, the IMU system cannot currently be considered clinically useful for lameness evaluation because of limitations in accuracy, attachment method, and lack of stance phase evaluation.
Abstract
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.
Abstract
Objective—To determine intralimb orientation changes with an inertial measurement unit (IMU) in hooves of horses at a walk and trot after induction of weight-bearing single forelimb lameness and to determine whether hoof orientations are similar to baseline values following perineural anesthesia.
Animals—6 clinically normal horses.
Procedures—3-D hoof orientations were determined with an IMU mounted on the right forelimb hoof during baseline conditions, during 3 grades of lameness (induced by application of pressure to the sole), and after perineural anesthesia. Linear acceleration profiles were used to segment the stride into hoof breakover, stance, initial swing, terminal swing, and total swing phases. Intralimb data comparisons were made for each stride segment. A repeated-measures mixed-model ANOVA was used for data analysis.
Results—Lameness resulted in significant changes in hoof orientation in all planes of rotation. A significant increase in external rotation and abduction and a significant decrease in sagittal plane rotation of the hoof were detected at hoof breakover during lameness conditions. For sagittal plane orientation data, the SDs determined following perineural anesthesia were higher than the SDs for baseline and lameness conditions.
Conclusions and Clinical Relevance—Results of this study indicated the IMU could be used to detect 3-D hoof orientation changes following induction of mild lameness at a walk and trot. An increase in data variability for a sagittal orientation may be useful for assessment of local anesthesia for hooves. The IMU should be further evaluated for use in clinical evaluation of forelimb lameness in horses.
Abstract
Objective—To evaluate the effect of underwater treadmill exercise on static postural sway in horses with experimentally induced carpal joint osteoarthritis under various stance conditions.
Animals—16 horses.
Procedures—On day 0, osteoarthritis was induced arthroscopically in 1 randomly selected middle carpal joint of each horse. Beginning on day 15, horses were assigned to either underwater or overground (without water) treadmill exercise at the same speed, frequency, and duration. Two serial force platforms were used to collect postural sway data from each horse on study days −7, 14, 42, and 70. Horses were made to stand stationary on the force platforms under 3 stance conditions: normal square stance, base-narrow placement of the thoracic limbs, and removal of visual cues (blindfolded) during a normal square stance. The mean of 3 consecutive, 10-second trials in each condition was calculated and used for analysis.
Results—Displacement of the center of pressure differed significantly depending on the stance condition. Among horses exercised on the underwater treadmill, postural stability in both the base-narrow and blindfolded stance conditions improved, compared with findings for horses exercised on the overground treadmill. Horses exercised on the overground treadmill were only successful at maintaining a stable center of pressure during the normal square stance position.
Conclusions and Clinical Relevance—Variations in stance position had profound effects on the mechanics of standing balance in horses with experimentally induced carpal joint osteoarthritis. Underwater treadmill exercise significantly improved the horses’ postural stability, which is fundamental in providing evidence-based support for equine aquatic exercise.
Abstract
OBJECTIVE
To assess whether the combination of hyaluronan, sodium chondroitin sul-fate, and N-acetyl-d-glucosamine (HCSG) lubricates articular cartilage in vitro and modulates joint lubrication in vivo.
ANIMALS
16 healthy adult horses.
PROCEDURES
The effects of HCSG injections on SF lubricant properties and joint health, immediately after injury and 2 weeks later, were analyzed by use an equine osteochondral fracture model of post-traumatic osteoarthritis (OA). Middle carpal joints of adult horses were randomly assigned to 1 of 4 surgical treatment groups as follows: normal nonsurgical group (n = 8), normal sham-surgical group (8), OA-induced surgical group with HCSG injection (8), or OA-induced surgical group with saline (0.9% NaCl) solution injection (8). Synovial fluid was aspirated periodically and analyzed for boundary lubrication function and lubricant molecules. At 17 days, joints were screened for gross pathological changes.
RESULTS
Induction of OA led to an impairment of SF lubrication function and diminished hyaluronan concentration in a time-dependent manner following surgery, with HCSG injection lessening these effects. Certain friction coefficients approached those of unaffected normal equine SF. Induction of OA also caused synovial hemorrhage at 17 days, which was lower in joints treated with HCSG.
CONCLUSIONS AND CLINICAL RELEVANCE
After induction of OA, equine SF lubricant function was impaired. Hyaluronan-sodium chondroitin sulfate–N-acetyl-d-glucosamine injection restored lubricant properties at certain time points and reduced pathological joint changes.
Abstract
Objective—To develop an antibody that specifically recognizes collagenase-cleaved type-II collagen in equine articular cartilage.
Sample Population—Cartilage specimens from horses euthanatized for problems unrelated to the musculoskeletal system.
Procedure—A peptide was synthesized representing the carboxy- (C-) terminus (neoepitope) of the equine type-II collagen fragment created by mammalian collagenases. This peptide was used to produce a polyclonal antibody, characterized by western analysis for reactivity to native and collagenase-cleaved equine collagens. The antibody was evaluated as an antineoepitope antibody by ELISA, using peptides ± an amino acid at the C-terminus of the immunizing peptide. Collagen cleavage was assayed from equine articular cartilage cultured with interleukin-1 (IL-1), ± a synthetic MMP inhibitor, BAY 12-9566. Cartilage specimens from osteoarthritic and nonarthritic joints were compared for antibody staining.
Results—An antibody, 234CEQ, recognized only collagenase- generated 3/4-length fragments of equine type-II collagen. This was a true antineoepitope antibody, as altering the C-terminus of the immunizing peptide significantly decreased competition for binding in an inhibition ELISA. The IL-1-induced release of type-II collagen fragments from articular cartilage was prevented with the MMP inhibitor. Cartilage from an osteoarthritic joint of a horse had increased staining with the 234CEQ antibody, compared with normal articular cartilage.
Conclusions and Clinical Relevance—We generated an antineoepitope antibody recognizing collagenase- cleaved type-II collagen of horses. This antibody detects increases in type-II collagen cleavage in diseased equine articular cartilage. The 234CEQ antibody has the potential to aid in the early diagnosis of arthritis and to monitor treatment responses. (Am J Vet Res 2001;62:1031–1039)
Abstract
OBJECTIVE To determine the effects of altering location of right forelimb and pelvic sensors on kinematic data obtained with a commonly used inertial sensor system during gait analysis of trotting horses.
DESIGN Experimental study.
ANIMALS 12 horses with mild to moderate lameness of at least 1 hind limb, with or without lameness of the forelimbs.
PROCEDURES All horses were examined while trotting on a high-speed treadmill. The right forelimb sensor was tested at 3 anatomic locations in random order: dorsal midline and 2 cm medial and lateral to that midline. During another treadmill session, the pelvic sensor was tested at 5 anatomic locations in random order: dorsal midline, 2 cm to the right and left of midline, and 2 cm cranial and caudal to the tubera sacrale on the midline. Laterality of the pelvic sensor was analyzed in 2 ways: sensor toward the right or left and sensor toward or away from the lame or lamest hind limb. Maximum and minimum differences in head and pelvic motion and vector sum values were ranked and compared with values for the midline location by means of mixed-model ANOVA.
RESULTS Altering the location of the right forelimb sensor by 2 cm medially or laterally had no significant effect on forelimb or hind limb kinematics. However, location of the pelvic sensor had a significant effect on minimum difference in pelvic motion, regardless of whether the data were analyzed by laterality (right vs left) or toward versus away from the lame hind limb.
CONCLUSIONS AND CLINICAL RELEVANCE Results of this study indicated that a 2-cm change in the location of the pelvic sensor during kinematic gait analysis had a significant effect on hind limb kinematic data of the system used. Therefore, placement of this sensor needs to be anatomically accurate.
Abstract
Objective—To determine whether muscle moment arms at the carpal and metacarpophalangeal joints can be modeled as fixed-radius pulleys for the range of motion associated with the stance phase of the gait in equine forelimbs.
Sample Population—4 cadaveric forelimbs from 2 healthy Thoroughbreds.
Procedure—Thin wire cables were sutured at the musculotendinous junction of 9 forelimb muscles. The cables passed through eyelets at each muscle's origin, wrapped around single-turn potentiometers, and were loaded. Tendon excursions, measured as the changes in lengths of the cables, were recorded during manual rotation of the carpal (180° to 70°) and metacarpophalangeal (220° to 110°) joints. Extension of the metacarpophalangeal joint (180° and 220°) was forced with an independent loading frame. Joint angle was monitored with a calibrated potentiometer. Moment arms were calculated from the slopes of the muscle length versus joint angle curves.
Results—At the metacarpophalangeal joint, digital flexor muscle moment arms changed in magnitude by ≤ 38% during metacarpophalangeal joint extension. Extensor muscle moment arms at the carpal and metacarpophalangeal joints also varied (≤ 41% at the carpus) over the range of joint motion associated with the stance phase of the gait.
Conclusions and Clinical Relevance—Our findings suggest that, apart from the carpal flexor muscles, muscle moment arms in equine forelimbs cannot be modeled as fixed-radius pulleys. Assuming that muscle moment arms at the carpal and metacarpophalangeal joints have constant magnitudes may lead to erroneous estimates of muscle forces in equine forelimbs. (Am J Vet Res 2003;64:351–357)