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Abstract

Objectives—To establish reference mechanical nociceptive threshold (MNT) values of the equine thoracic limb and to assess the use of MNT values to detect pain associated with induced osteoarthritis in the middle carpal joint.

Animals—24 adult horses.

Procedures—MNT values were evoked by a pressure algometer at 17 sites within each thoracic limb during 2 baseline sessions conducted an average of 5 days apart. Effects of age, sex, weight, and wither height on MNT values were assessed separately for each site. Tolerance of horses to the procedure was graded subjectively and correlated with MNT values. Synovitis and osteoarthritis were induced arthroscopically in the middle carpal joint of 1 randomly selected thoracic limb. The opposite limb served as a sham-operated control limb. Mechanical nociceptive threshold values were recorded weekly and correlated with clinical, radiographic, and necropsy scores measured over 10 weeks. Lower MNT values corresponded with increased pain, whereas higher MNT values indicated reduced pain.

Results—A gradual increase in MNT values was detected from proximal-to-distal sites of the thoracic limbs. High MNT values were recorded for geldings and tall horses. In general, tolerance to procedure scores was positively correlated with overall pooled MNT values within each thoracic limb. From 2 to 6 weeks after surgery, the osteoarthritic limb had significantly reduced MNT values within the carpal region. The osteoarthritic limb also had significant changes in clinical examination, radiographic, and necropsy scores, which were poorly correlated with MNT values.

Conclusions and Clinical Relevance—Pressure algometry provided objective assessment of nociception of the thoracic limb; however, MNT values were poorly correlated with clinical variables used to assess osteoarthritis.

Full access
in American Journal of Veterinary Research

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.

Full access
in American Journal of Veterinary Research

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.

Full access
in American Journal of Veterinary Research

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.

Full access
in American Journal of Veterinary Research

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.

Full access
in American Journal of Veterinary Research

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.

Full access
in American Journal of Veterinary Research

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.

Full access
in Journal of the American Veterinary Medical Association

Abstract

Objective

To determine whether serum or synovial fluid concentrations of chondroitin sulfate epitope 846 and carboxy propeptides of type II collagen (CPII) can be used to diagnose osteochondral fragmentation (OC) in horses.

Animals

38 horses with unilateral OC of the radiocarpal (n = 31) or intercarpal (33) joints and 8 clinically and radiographically normal horses.

Procedures

For horses with OC, serum and synovial fluid concentrations of epitope 846, CPII, and keratan sulfate (KS) were determined, along with synovial fluid WBC counts and total protein concentrations. Serum epitope 846, CPII, and KS concentrations were measured in control horses.

Results

Synovial fluid epitope 846 and total protein concentrations were significantly higher in the joints with OC than in unaffected joints, but CPII and KS concentrations and WBC counts were not. Synovial fluid total protein and 846 epitope concentrations were linearly related to grade of OC. Serum epitope 846 and CPII concentrations were significantly higher in horses with OC than in control horses. Discriminant analysis allowed 27 of 34 (79%) horses to be correctly classified as having or not having OC on the basis of serum epitope 846 and CPII concentrations.

Conclusions

Results suggest that serum and synovial fluid concentrations of epitope 846 and CPII are associated with OC. Increases in concentrations of epitope 846 and CPII suggest that increased synthesis of cartilage aggrecan and type II procollagen may be associated with OC.

Clinical Relevance

Measurement of serum epitope 846 and CPII concentrations may be useful in the diagnosis of OC in horses. (Am J Vet Res 1999;60:306–309)

Free access
in American Journal of Veterinary Research

SUMMARY

The use of periosteal autografts to resurface osteochondral defects was investigated in 10 horses (2 to 3 years old), and the repair tissue was characterized morphologically. Middle carpal joint arthrotomies were made, and osteochondral defects were induced bilaterally on the distal articular surface of each radial carpal bone. Each defect measured approximatively 1 cm2 and extended 3 mm into the subchondral bone plate. Residual subchondral bone plate of control and principal defects was perforated by drilling. A sterile fibrin adhesive was made by mixing a fibrinogen component and a thrombin component. A periosteal autograft was harvested from the proximal portion of the tibia and was glued onto the recipient osseous surface, with its cambium facing the joint cavity. Control defects were glued, but not grafted. Horses were walked 1 hour daily on a walker, starting at postoperative week 7 and continuing for 9 weeks. Sixteen weeks after the grafting procedure was done, carpal radiography was performed, after which horses were euthanatized. Quality of repair tissue of control and grafted defects was evaluated and compared grossly, histologically, and histochemically. Using a reticule, the proportions of various repair tissue types filling each defect were quantitated.

Seven weeks after the grafting procedure was done, bilateral arthroscopy revealed synovial adhesions and marginal pannus formation in control and grafted defects. None of the autografts was found floating unattached within the respective middle carpal joints. At 16 weeks, the gross appearance of most grafted and nongrafted defects was similar, and repair was dominated by a fibrous pannus. In 4 grafted defects, bone had formed either concentrically within the defect or eccentrically in the fibrous adhesions between the defect and the joint margin. Histologically, all grafted and nongrafted defects were repaired similarly by infiltration of a mixture of fibrous tissue, fibrocartilage, and bone. Fibrous tissue was the predominant tissue in most defects and its mean proportion was 56 and 59% in the grafted and nongrafted defects, respectively. Fibrocartilaginous tissue in the deeper layers approximated 20%, and woven bone at the base of the defect was 20% in all defects. Histochemically, difference in staining for proteoglycans was not observed between grafted and nongrafted defects. Little remaining original periosteal graft tissue was evident at the defect sites. The only distinguishing feature of grafted defects was the presence of islands of bone formation either at the defect site (n = 2 horses), or in somewhat dorsally displaced tissue that was incorporated in fibrous adhesions (n = 2 horses). It was concluded that use of periosteal autografts did not improve the healing of osteochondral defects of the distal portion of the radial carpal bone. The repair tissue produced in grafted and nongrafted defects was similar and was principally fibrous in nature.

Free access
in American Journal of Veterinary Research

Summary

The effects of intra-articular administration of methylprednisolone acetate (mpa) on the healing of full-thickness osteochondral defects and on normal cartilage were evaluated in 8 horses. In group-1 horses (n = 4), a 1-cm-diameter, full-thickness defect was created bilaterally in the articular cartilage on the dorsal distal surface of the radial carpal bone. Cartilage defects were not created in group-2 horses (n = 4). One middle carpal joint was randomly selected in each horse (groups 1 and 2), and treated with an intra-articular injection of 100 mg of mpa, once a week for 4 treatments. Injections began 1 week after surgery in group-1 horses. The contralateral middle carpal joint received intra-articular injections of an equivalent volume of 0.9% sodium chloride solution (scs), and served as a control. Horses were evaluated for 16 weeks, then were euthanatized, and the middle carpal joints were examined and photographed. Synovial and articular cartilage specimens were obtained for histologic and histochemical evaluation.

Gross morphometric evaluation of the healing defects in group-1 horses revealed that 48.6% of the defect in control joints and 0% of the defect in mpa-treated joints was resurfaced with a smooth, white tissue, histologically confirmed as fibrocartilage. This replacement tissue was a firmly attached fibrocartilage in control joints and a thin fibrous tissue in mpa-treated joints. The articular cartilage in joints treated with mpa had morphologic changes, including chondrocyte cluster formation, loss of palisading architecture, and cellular necrosis in both groups of horses. Histochemical (safranin-0) staining intensity was reduced significantly (P < 0.05) in all layers of articular cartilage in mpa-treated joints in groups 1 and 2. In the replacement tissue, intense safranin-0 staining was found only in the chondrocyte clusters deep in the tissue of control joints, confirming fibrocartilage repair. Intra-articular administration of mpa in this dosing regimen thus induced degenerative changes in normal articular cartilage and resulted in histomorphologic changes in the repair of full-thickness articular osteochondral defects in horses.

Free access
in American Journal of Veterinary Research