Effects of intra-articular administration of methylprednisolone acetate on normal equine articular cartilage

The effects of the corticosteroid 6-alpha-methylprednisolone acetate on normal equine articular cartilage were evaluated, using the middle carpal joint in 4 clinically normal young horses. One middle carpal joint of each horse was injected 3 times with 100 mg of 6-alpha-methylprednisolone acetate, at 14-day intervals. The opposite middle carpal joint (control) was injected with 2.5 ml of lactated Ringer solution at the same intervals. Effects were studied until 8 weeks after the first injection. Evaluation included clinical and radiographic examination, and gross, microscopic, and biochemical evaluation of joint tissues. Horses remained clinically normal during the study, and significant radiographic changes were not observed. Safranin-0 matrix staining intensity and uronic acid content were significantly (P less than 0.05) lower and hydroxyproline content was significantly (P less than 0.05) higher in articular cartilage of corticosteroid-injected joints vs control joints.     

Clinical effects of exercise on subchondral bone of carpal and metacarpophalangeal joints in horses

OBJECTIVE: To determine effects of treadmill exercise on subchondral bone of carpal and metacarpophalangeal joints of 2-year-old horses. ANIMALS: 12 healthy 2-year-old horses. PROCEDURE: Horses were randomly assigned to the control (n = 6) or exercised (6) groups. Horses in the exercised group ran on a high-speed treadmill 5 d/wk for 6 months. Horses in the control group were hand walked for the same amount of time. Results of clinical, radiographic, nuclear scintigraphic, and computed tomographic examinations, and serum and synovial concentrations of biochemical markers of bone metabolism were compared between groups. RESULTS: Exercised horses were significantly lamer at the end of the study than control horses. Radionuclide uptake in the metacarpal condyles, but not in the carpal joints, was greater in exercised horses, compared with control horses. Exercised horses also had a higher subchondral bone density in the metacarpal condyles than control horses, but such differences were not detected in the carpal bones. CONCLUSIONS AND CLINICAL RELEVANCE: None of the diagnostic techniques evaluated was sufficiently sensitive to detect all osteochondral damage. Computed tomography and computed tomographic osteoabsorptiometry were superior to conventional radiography for detecting small osteochondral fragments. Nuclear scintigraphy was a sensitive indicator of subchondral bone change but lacked specificity for describing lesions and discerning normal bone remodeling from damage. Newer techniques such as computed tomography may help clinicians better diagnose early and subtle joint lesions in horses prior to development of gross joint damage.     

Synovial fluid and plasma kinetics of methylprednisolone and methylprednisolone acetate in horses following intra-articular administration of methylprednisolone acetate

Synovial fluid and plasma kinetics of methylprednisolone acetate (MPA) and methylprednisolone (MP) after a single intra-articular administration of MPA at a therapeutic dose (111 mg in toto) was measured in five horses. MPA was detected in synovial fluid for two to six days post injection and MP, which results from synovial MPA hydrolysis, was present in pharmacologically significant concentrations for 4.8 to 39 days, depending on the horse. MPA synovial concentration was maximal (289 +/- 284 micrograms/ml) at the first sampling time (2 h after administration) and MP synovial concentration was maximal (from 58.9 to 379.5 micrograms/ml) at the first or second sampling time (2 to 10 h after administration). Thereafter, both MP and MPA declined rapidly. From time of administration to about five days later, MP synovial fluid concentration fell progressively with a half-time of 9.95 h. Subsequently, the MP synovial fluid concentration decreased more slowly with an apparent half-time of 115 h. During the first 24 h following MPA administration, trace amounts of MP (less than 5 ng/ml) were detected in plasma. Plasma hydrocortisone levels were depressed for three to four days after administration but adrenal responsiveness to adrenocorticotrophic hormone tests remained unaffected.     

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.     

The dose-related effects of phenylbutazone and a methylprednisolone acetate formulation (Depo-Medrol®) on cultured explants of equine carpal articular cartilage

The dose-related effects of phenylbutazone and Depo-Medrol® on chondrocyte viability and chondrocyte-mediated synthesis and depletion of proteoglycans were investigated using cultured explants of equine middle carpal joint articular cartilage. Explants from 12 horses (941 × 3 mm diameter) were cultured for a total of 5 days, which included 3 days' exposure to either phenylbutazone (0, 2, 20, 200 or 2000 μg/mL) or Depo-Medrol (0, 20, 200 or 2000 μg/mL). For each explant, amino sugar content was used as a measure of proteoglycan content, ³⁵S incorporation as a measure of the rate of proteoglycan synthesis and the number of pyknotic nuclei as a measure of cell death. During culture, control explants remained metabolically active and viable but suffered a net loss of proteoglycans. Proteoglycan loss was reduced by the presence of either phenylbutazone or Depo-Medrol. This effect was significant at clinically relevant concentrations of phenylbutazone (2–20 μg/mL), but not Depo-Medrol (20–200 μg/mL). Depo-Medrol caused a dose-dependent suppression of proteoglycan synthesis at all concentrations, but chondrocyte viability was affected only at the 2000 μg/mL dose. Phenylbutazone affected proteoglycan synthesis and cell viability only at the 2000 μg/mL concentration. At all concentrations, the anticatabolic effects of each drug influenced the proteoglycan content of the explants far more than did any antianabolic or cytotoxic drug effect. The results suggest that the therapeutic potential of both phenylbutazone and Depo-Medrol may not be restricted to their anti-inflammatory effects on the soft tissues of the joint.     

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)     

Results of treatment of subchondral bone cysts in the medial condyle of the equine femur with an autogenous cancellous bone graft

The results of surgical treatment of 10 subchondral bone cysts, all located in the medial femoral condyle, are presented. A cancellous bone graft was used in nine cases and a two component acrylic bone cement was used in the case of an extremely large cyst. Eight out of the 10 cases made a satisfactory clinical recovery.     

The effect of compacted cancellous bone grafting on the healing of subchondral bone defects of the medial femoral condyle in horses

Objective— To compare the quality of second-intention healing and that of compacting sternally harvested cancellous bone into subchondral bone defects of the medial femoral condyle in horses.
Study Design— A controlled experiment using a surgical technique that minimizes soft tissue trauma, customized for consistency among horses.
Animals or Sample Population— Ten horses, aged 2 to 5 years, free of hindlimb lameness and with radiographically normal stifles.
Methods— After a 12.7-mm-diameter × 19-mm-deep defect was created into randomly selected medial femoral condyles, bone and cartilage healing was evaluated over a 6-month period in control horses (  n = 5  ) and horses receiving a compacted cancellous bone graft (  n = 5  ). Healing was evaluated using lameness assessment, radiographic and microradiographic interpretation, arthroscopic appearance, percent bone fill, proteoglycan content, and histology.
Results— Six months after surgery, there was no significant difference between grafted and ungrafted defects with respect to lameness, radiographic score, or percent bone fill. Histologically, grafted defects were characterized by the presence of dead graft and secondary cyst formation in four defects. Ungrafted defects filled with fibrous tissue and no cyst formation were identified.
Conclusions— Grafted defects do not heal better than ungrafted defects, and lameness was not affected by surgical technique.
Clinical Significance— Cartilage healing is similar in grafted and ungrafted defects in the equine medial femoral condyle at 6 months, suggesting that surgical debridement alone of cystic structures remains the treatment of choice.     

Changes in articular cartilage after intra-articular injections of methylprednisolone acetate in horses

Eight mature horses with no prior signs of joint disease or history of intra-articular therapy were treated with 8 weekly intra-articular injections of methylprednisolone acetate. Treatments were given at a dose of 120 mg/joint into the right radiocarpal and intercarpal joints, with the left joints as untreated controls. Articular cartilage samples were obtained at necropsy 1, 4, and 8 weeks after the last injection. Compared with controls, cartilage from injected joints had a loss of hematoxylin basophilia and decreased intensity of staining in safranin O fast green dye. Chondrocyte necrosis and hypocellularity were observed in all samples of cartilage from treated joints. Proteoglycan content and its rate of synthesis were reduced. There was a progressive loss of proteoglycan content, whereas proteoglycan synthesis increased somewhat 4 and 8 weeks after treatment. Collagen content was unchanged, but its rate of synthesis was markedly inhibited. Collagen synthesis did not recover, but remained decreased at 5 to 15% of the values from untreated cartilage. Water percentage was increased, but fibronectin content was not significantly different. A single injection of methylprednisolone acetate was also given into the right metacarpophalangeal joints of 3 of the 8 horses in this group, with the left joints serving as untreated controls. Sixteen weeks after the treatment, cartilage of the treated joints had a loss of histochemical staining and proteoglycan content was reduced to 50% of control values. The mean rate of proteoglycan synthesis and mean fibronectin content were increased, but the differences were not statistically significant (P greater than 0.05). Other variables were essentially unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of methylprednisolone acetate and glucosamine on proteoglycan production by equine chondrocytes in vitro

OBJECTIVE: To evaluate the effects of methylprednisolone acetate (MPA) on proteoglycan production by equine chondrocytes and to investigate whether glucosamine hydrochloride modulates these effects at clinically relevant concentrations. SAMPLE POPULATION: Articular cartilage with normal gross appearance from metacarpophalangeal and metatarsophalangeal joints of 8 horses (1 to 10 years of age). PROCEDURES: In vitro chondrocyte pellets were pretreated with glucosamine (0, 1, 10, and 100 microg/mL) for 48 hours and exposed to MPA (0, 0.05, and 0.5 mg/mL) for 24 hours. Pellets and media were assayed for proteoglycan production (Alcian blue precipitation) and proteoglycan content (dimethylmethylene blue assay), and pellets were assayed for DNA content. RESULTS: Methylprednisolone decreased production of proteoglycan by equine chondrocytes at both concentrations studied. Glucosamine protected proteoglycan production at all 3 concentrations studied. CONCLUSIONS AND CLINICAL RELEVANCE: Methylprednisolone, under noninflammatory conditions present in this study, decreased production of proteoglycan by equine chondrocytes. Glucosamine had a protective effect against inhibition of proteoglycan production at all 3 concentrations studied. This suggested that glucosamine may be useful as an adjunct treatment when an intra-articular injection of a corticosteroid is indicated and that it may be efficacious at concentrations relevant to clinical use.     

Qualitative aspects of the incorporation of equine cancellous bone grafts

The incorporation of autogenous cancellous bone graft was studied in eight yearling ponies. The site for the defect to be grafted was chosen so that the effect on the graft, of both the host cortical and trabecular bone, could be assessed. To obtain information concerning the vitality of the graft and the dynamic aspects of the modelling and remodelling processes of graft incorporation, a double and treble tetracycline intravital labelling technique was used. Radiographs of the graft and host tissues of all ponies were obtained regularly, but were of little assistance in assessing graft incorporation. The ponies were destroyed humanely at regular intervals between nine and 241 days after installation of the graft, followed by histological examination of undecalcified sections. The study revealed that all installed graft trabeculae showed signs of non-vitality at nine days after installation and gradually disintegrated. Two processes of new bone formation were observed. First, finger-like projections of immature new trabeculae were found to originate from the graft/host interfaces. Second, a gradual process of accretion of osteoid and woven bone upon disintegrating graft trabeculae occurred uniformly throughout the graft. The graft adapted to the structure of opposing host bone by corticalisation and trabecularisation. The present study confirmed clinical observations relating to convalescence time following grafting of large osseous defects in horses and indicated that equine bone reacts to autogenous bone grafts in a similar manner to other mammals.     

Influence of intensity and changes of physical activity on bone mineral density of immature equine subchondral bone

Reasons for performing study: Subchondral bone provides structural support to overlying articular cartilage and plays an important biomechanical role in osteochondral diseases. Mechanical features of bone correlate strongly with bone mineral density, which is directed by the loading conditions to which the tissue is subjected. Objective: To investigate the influence of physical activity levels on subchondral bone mineral density (sBMD) in foals during early development. Methods: Three groups of foals were subjected to different physical activity levels from birth until age 5 months. A proportion of these foals were subjected to euthanasia at 5 months while remaining foals were subjected to similar physical activity levels for 6 months until euthanasia at 11 months. Osteochondral specimens were collected for measurement of sBMD with peripheral quantitative computed tomography at 2 differently loaded anatomical sites of the proximal phalangeal bone at 1, 2, 3, 4 and 5 mm depth from the osteochondral junction. Results: Growth significantly increased sBMD but by a different amount depending on anatomical location and physical activity level. Significantly higher sBMD was found at the habitually loaded central area in comparison to the intermittently peak loaded marginal site. Exercise increased sBMD throughout the whole depth of analysed tissue, but changes were generally more obvious at a depth of 2 mm. Interestingly, foals subjected to additional sprint training preserved the exercise‐induced sBMD increase at the habitually loaded central area during the 6 months of the second phase of the study. Conclusions: Habitual low‐intensity loading elicits a greater response in sBMD in quantitative terms than high‐intensity low‐frequency loading at the sites investigated in this study. Future sBMD may be influenced by means of well‐tailored exercise regimens at young age. Potential relevance: Specific physical activity levels during early development may potentially reduce the prevalence of osteochondral injury later in life.     

Effects of sodium hyaluronate and methylprednisolone acetate on proteoglycan synthesis in equine articular cartilage explants

OBJECTIVE: To determine effects of sodium hyaluronate (HA) on corticosteroid-induced cartilage matrix catabolism in equine articular cartilage explants. SAMPLE POPULATION: 30 articular cartilage explants from fetlock joints of 5 adult horses without joint disease. PROCEDURE: Articular cartilage explants were treated with control medium or medium containing methylprednisolone acetate (MPA; 0.05, 0.5, or 5.0 mg/mL), HA (0.1, 1.0, or 1.5 mg/mL), or both. Proteoglycan (PG) synthesis was measured by incorporation of sulfur 35-labeled sodium sulphate into PGs, and PG degradation was measured by release of radiolabeled PGs into the medium. Total glycosaminoglycan (GAG) content in media and explants and total explant DNA were determined. RESULTS: Methylprednisolone acetate caused a decrease in PG synthesis, whereas HA had no effect. Only the combination of MPA at a concentration of 0.05 mg/mL and HA at a concentration of 1.0 mg/mL increased PG synthesis, compared with control explants. Methylprednisolone acetate increased degradation of newly synthesized PGs into the medium, compared with control explants, and HA alone had no effect. Hyaluronate had no effect on MPA-induced PG degradation and release into media. Neither MPA alone nor HA alone had an effect on total cartilage GAG content. Methylprednisolone acetate caused an increase in release of GAG into the medium at 48 and 72 hours after treatment. In combination, HA had no protective effect on MPA-induced GAG release into the medium. Total cartilage DNA content was not affected by treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Our results indicate that HA addition has little effect on corticosteroid-induced cartilage matrix PG catabolism in articular cartilage explants.     

The effect of exercise and conditioning on equine red blood cell characteristics

The effect of exercise and conditioning on 2,3-diphosphoglycerate levels was studied in nine mature horses. During a 12 minute exercise bout producing heart rates of 165 bpm, 2,3-DPG was significantly increased (p<.05). In addition, exercising levels of 2,3-DPG were increased (p<.05) approximately 8% after a six-week submaximal conditioning program. These increases could not be entirely attributed to changes in erythrocyte number. Mean corpuscular volume was also increased during exercise (p<.05) but was not altered by conditioning.     

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.     

Preliminary study of quantitative aspects and the effect of pulsed electromagnetic field treatment on the incorporation of equine cancellous bone grafts

The quantitative aspects of equine cancellous bone graft incorporation and the possibility of influencing graft incorporation by daily exposure to a pulsed electromagnetic field (PEMF) was studied in eight yearling ponies. In order to be able to quantify formative aspects of graft remodelling, a double and treble tetracycline intravital labelling technique was used. Intravital radiographs were obtained at regular intervals throughout the trial, but were found to be of little assistance in assessing any differences between stimulated and non-stimulated grafts. The ponies were humanely destroyed at regular intervals between nine and 241 days after installation of the graft. Light microscopy and fluorescent light microscopy were used to evaluate quantitative aspects of graft incorporation and to compare PEMF-stimulated grafts with control grafts. There was a small but statistically significant effect of PEMF-stimulation on cancellous bone graft incorporation. In view of this, these observations can only be considered as indicative of a possible trend, but should encourage further studies using different signal modalities.     

Pharmacokinetics of methylprednisolone acetate after intra-articular administration and subsequent suppression of endogenous hydrocortisone secretion in exercising horses

Objective-To determine the pharmacokinetics of methylprednisolone (MP) and the relationship between MP and hydrocortisone (HYD) concentrations in plasma and urine after intra-articular (IA) administration of 100 or 200 mg of MP acetate (MPA) to horses. Animals-Five 3-year-old Thoroughbred mares. Procedures-Horses exercised on a treadmill 3 times/wk during the study. Horses received 100 mg of MPA IA, then 8 weeks later received 200 mg of MPA IA. Plasma and urine samples were obtained at various times for 8 weeks after horses received each dose of MPA; concentrations of MP and HYD were determined. Pharmacokinetic-pharmacodynamic estimates for noncompartmental and compartmental parameters were determined. Results-Maximum concentration of MP in plasma was similar for each MPA dose; concentrations remained greater than the lower limit of quantitation for 18 and 7 days after IA administration of 200 and 100 mg of MPA, respectively. Maximum concentration and area under the observed concentration-time curve for MP in urine were significantly higher (approximately 10-and 17-fold, respectively) after administration of 200 versus 100 mg of MPA. Hydrocortisone concentration was below quantifiable limits for ≥ 48 hours in plasma and urine of all horses after administration of each MPA dose. Conclusions and Clinical Relevance-Pharmacokinetics of MP may differ among IA MPA dosing protocols, and MP may be detected in plasma and urine for a longer time than previously reported. This information may aid veterinarians treating sport horses. Further research is warranted to determine whether plasma HYD concentration can aid identification of horses that received exogenous glucocorticoids.     

The effect of training on equine metacarpal bone breaking strength

Right third metacarpal bones (n=24) from Thoroughbreds, 24 to 48 months old and in race training, were tested to failure in 3 point bending. The neutral load axis was estimated and the distance from the axis to the outer dorsal cortical surface measured. Mid-diaphyseal dorsopalmar and lateromedial outer cortical and medullary diameters were measured. Breaking strength, cortical area and area moment of inertia were also calculated. Significant correlations were demonstrated between months in training and dorsopalmar bone diameter, cortical area and area moment of inertia. Significant linear models were illustrated between the same 3 variables and number of months in training. It was concluded that increased duration of training significantly enlarges dorsopalmar bone diameter, cortical area and area moment of inertia. Training did not affect metacarpal bone breaking strength as determined by 3 point bending.     

Validation of magnetic resonance imaging for measurement of equine articular cartilage and subchondral bone thickness

OBJECTIVE: To validate use of magnetic resonance images (MRIs) for measurement of equine articular cartilage and subchondral bone thickness by comparison with measurements in histologic specimens. SAMPLE POPULATION: 32 cadaveric carpal joints from 16 horses. PROCEDURE: Magnetic resonance imaging was performed by use of 3-dimensional fast spoiled gradient echo (SPGR) and T2* 3-dimensional fast gradient echo (GRE) pulse sequences with and without fat saturation. Standard sites on the medial and lateral facets of the intermediate, radial, and third carpal bones were used for subchondral bone and articular cartilage thickness measurements. Digital image analysis software was used for MRI measurements 10 mm from the dorsal extent and perpendicular to the articular surface. Histomorphometric measurements of hyaline, calcified cartilage, and subchondral bone thickness were obtained at selected sites. Comparisons between histomorphometric and MRI measurements and between magnetic resonance pulse sequences were evaluated. RESULTS: There were significant correlations between GRE and SPGR and SPGR and histologic measurements of articular cartilage, with no significant difference between measurements and good agreement. When calcified cartilage was excluded from the histologic measurement, MRI measurements were significantly greater than histologic measurements. For subchondral bone thickness, there was significant correlation between GRE and SPGR but GRE was significantly greater than SPGR measurements. Histomorphometric and MRI measurements were strongly correlated and not significantly different. CONCLUSIONS AND CLINICAL RELEVANCE: Magnetic resonance imaging provides a good representation of cartilage and subchondral bone thickness, supporting its use in the study and clinical diagnosis of osteochondral structure and alteration.