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Abstract

Objective—To investigate effects of osteochondral injury on high-mobility group box chromosomal protein 1 (HMGB-1) concentrations in synovial fluid (SF) from Thoroughbreds and to compare these results with radiographic and arthroscopic scores of severity of joint injury.

Animals—40 clinically normal rested Thoroughbreds (group 1) and 45 Thoroughbreds with osteochondral injury as a result of racing.

Procedures—SF was obtained from the metacarpophalangeal (MCP) joints, metatarsophalangeal (MTP) joints, middle carpal joints, and radiocarpal joints. For group 2, radiographic and arthroscopic scores were determined. Concentrations of SF HMGB-1 were determined by use of an ELISA.

Results—SF HMGB-1 concentrations in osteochondral-injured MCP-MTP joints were significantly higher than in normal MCP-MTP joints. Similarly, SF HMGB-1 concentrations in osteochondral-injured carpal joints were significantly higher than in normal carpal joints. Radiographic and arthroscopic scores were not correlated with SF HMGB-1 concentrations. Synovial fluid HMGB-1 concentrations ≥ 11 ng/mL for MCP-MTP joints and ≥ 9 ng/mL for carpal joints discriminated osteochondral-injured joints from normal joints. Horses with HMGB-1 concentrations ≥ 11 ng/mL for MCP-MTP joints were twice as likely to have an osteochondral injury, and horses with HMGB-1 concentrations ≥ 9 ng/mL for carpal joints were 4 times as likely to have an osteochondral injury.

Conclusions and Clinical Relevance—Osteochondral injury was associated with a significant increase in SF HMGB-1 concentrations in MCP-MTP and carpal joints, compared with results for clinically normal Thoroughbreds. Analysis of SF HMGB-1 concentrations may be useful for evaluation of joint injury in horses.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To investigate the effects of exercise and osteochondral injury on concentrations of carboxy-terminal telopeptide fragments of type II collagen (CTX-II) in synovial fluid (SF) and serum of Thoroughbred racehorses and to compare findings with radiographic and arthroscopic scores of joint injury severity.

Animals—78 Thoroughbreds with (n = 38) and without (40) osteochondral injury.

Procedures—Serum and metacarpophalangeal or carpal joint SF samples were collected from noninjured horses before and at the end of 5 to 6 months of race training (pre- and postexercise samples, respectively) and from horses with osteochondral injury (1 joint assessed/horse). Synovial fluid and serum CTX-II concentrations were determined by use of an ELISA. Radiographic and arthroscopic scores of joint injury severity were determined for the injured horses.

Results—The CTX-II concentrations in SF and SF:serum CTX-II ratio were significantly higher for horses with joint injuries, compared with pre- and postexercise findings in noninjured horses. Serum CTX-II concentrations in postexercise and injured-horse samples were significantly lower than values in pre-exercise samples. On the basis of serum and SF CTX-II concentrations and SF:serum CTX-II ratio, 64% to 93% of serum and SF samples were correctly classified into their appropriate group (pre-exercise, postexercise, or injured-joint samples). In horses with joint injuries, arthroscopic scores were positively correlated with radiographic scores, but neither score correlated with SF or serum CTX-II concentration.

Conclusions and Clinical Relevance—Results suggested that serum and SF CTX-II concentrations and SF:serum CTX-II ratio may be used to detect cartilage degradation in horses with joint injury.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine the effects of horse age, osteochondral injury, and joint type on a synthesis biomarker and 3 degradative biomarkers of type II collagen in Thoroughbreds.

Animals—Healthy rested adult (3- to 12-year-old) Thoroughbreds (n = 19), yearling (1- to 2-year-old) Thoroughbreds (40), and Thoroughbred racehorses (2 to 7 years old) undergoing arthroscopic surgery for removal of osteochondral fragments that resulted from training or racing (41).

Procedures—Samples of blood and metacarpophalangeal, metatarsophalangeal, or carpal joint synovial fluid (SF) were collected from all horses. Commercially available assays were used to analyze SF and serum concentrations of type II collagen biomarkers of synthesis (carboxy propeptide of type II collagen [CPII]) and degradation (cross-linked C-telopeptide fragments of type II collagen [CTX II], neoepitope generated by collagenase cleavage of type I and II collagen [C1,2C], and neoepitope generated by collagenase cleavage of type II collagen [C2C]).

Results—Osteochondral injury affected concentrations of CPII, CTX II, C1,2C, and C2C in SF, serum, or both, compared with concentrations in healthy adult horses. Compared with adult horses, yearling horses had increased SF or serum concentrations of degradative biomarkers (CTX II, C1,2C, and C2C). Concentrations were higher in carpal than metacarpophalangeal or metatarsophalangeal joints for all biomarkers in osteochondral-injured horses. Variable differences in SF concentrations between joint types were detected in healthy adult and yearling horses.

Conclusions and Clinical Relevance—Horse age, osteochondral injury, and joint type all significantly affected type II collagen biomarker concentrations in SF and serum of Thoroughbreds.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine whether stromal cell-derived factor-1 (SDF-1) concentrations in serum, plasma, and synovial fluid differed among untrained, race-trained, and osteochondral-injured Thoroughbred racehorses.

Animals—22 racehorses without osteochondral injury and 37 racehorses with osteochondral injury.

Procedures—Horses without osteochondral injury were examined before and after 5 to 6 months of race training. Horses with osteochondral injury were undergoing arthroscopic surgery for removal of osteochondral fragments from carpal or metacarpophalangeal or metatarsophalangeal joints (fetlock joints). Serum, plasma, and fetlock or carpal synovial fluid samples were obtained and analyzed for SDF-1 concentration by use of an ELISA.

Results—In horses with fetlock or carpal joint injury, mean synovial fluid SDF-1 concentrations were significantly higher, serum SDF-1 concentrations were significantly lower, and synovial fluid-to-serum SDF-1 ratios were significantly higher than in untrained and trained horses. Synovial fluid SDF-1 concentrations were not significantly different between trained and untrained horses. Plasma SDF-1 concentrations were not different among the 3 groups. Results obtained with serum, compared with synovial fluid and plasma, had better sensitivity for differentiating between osteochondral-injured horses and uninjured horses. In horses with fetlock joint osteochondral injury, serum SDF-1 concentrations were correlated with radiographic and arthroscopic inflammation scores, but not arthroscopic cartilage scores.

Conclusions and Clinical Relevance—Results suggested that serum SDF-1 concentrations were more sensitive than plasma and synovial fluid concentrations for detection of osteochondral injury in the fetlock or carpal joint of racehorses. Analysis of serum and synovial SDF-1 concentrations in horses with experimentally induced joint injury may help define the onset and progression of post-traumatic osteoarthritis and aid in the evaluation of anti-inflammatory treatments.

Full access
in American Journal of Veterinary Research

Summary

Each of 5 healthy mares was given 5 consecutive im injections of ceftiofur sodium (2 mg/kg of body weight; 50 mg/ml) at 12-hour intervals. Ceftiofur concentrations were measured serially in serum, synovial fluid, peritoneal fluid, and urine, and were measured in CSF and endometrial tissue after the fifth dose. Mean elimination rate constant was 0.354 ± 0.101 h−1 and elimination half-life was 2.49 ± 0.49 hour. Mean serum ceftiofur concentrations peaked approximately 1 hour after each injection. The highest mean ceftiofur concentration was 5.09 μg/ml at 1 hour after the fifth dose for serum, 3.02 μg/ml at 2 hours after the fifth dose for synovial fluid, and 3.23 μg/ml at 4 hours after the fifth dose for peritoneal fluid. Mean urine concentrations reached 15.72 μg/ml at 1 hour after the fifth dose. Ceftiofur was not detected in csf or endometrial tissue. None of the mares had adverse reactions to the drug.

Free access
in American Journal of Veterinary Research

Summary

Pharmacokinetic values for flunixin meglumine (1 mg/kg of body weight) and phenylbutazone (4 mg/kg) dosages were determined after a single iv injection with and without concurrent intragastric administration of probenecid (50 mg/kg) in 6 healthy mares. Significant difference was not apparent in the pharmacokinetic values of flunixin meglumine with and without concurrent probenecid administration. Significant (P ≤ 0.05) increase was evident in the 12-hour mean concentration of phenylbutazone (11.45 ± 1.66 μg/ml without probenecid; 14.56 ± 1.20 μg/ml with probenecid) along with significant (P ≤ 0.05) reduction in its volume of distribution at steady state associated with concurrent probenecid administration (218.6 ± 11.52 ml/kg without probenecid; 169.4 ± 9.25 ml/kg with probenecid).

Free access
in American Journal of Veterinary Research

Abstract

Objectives

To determine effects of aging on sulfation of chondroitin sulfate (CS) in articular cartilage and synovial fluid from normal equine middle carpal joints, and to determine whether CS compositional analysis can be used to assess alterations in proteoglycan turnover in degenerative cartilage obtained from horses with carpal osteochondral fractures.

Sample Population

Carpal articular cartilage and synovial fluid from 44 cadavers with normal joints and from 16 Thoroughbred racehorses during routine carpal arthroscopic surgery.

Procedure

After papain/chondroitinase digestion of cartilage, CS disaccharides (unsulfated disaccharide ΔDi0S, and monosulfated disaccharides ΔDi4S and ΔDi6S) were quantified by capillary zone electrophoresis. The CS was purified from synovial fluid chondroitinase digested, and analyzed. The CS nonreducing terminal residues, N-acetylgalactosamine (galNAc) or glucuronic acid adjacent to a 4-sulfated or 6-sulfated galNAc, were quantified.

Results

In cartilage, the ΔDi6S-to-ΔDi4S ratio increased with age; in degenerative cartilage, this ratio was not significantly different from the normal value. Percentage of ΔDi0S decreased with age and was significantly higher in degenerative than in normal cartilage. The galNAc4S and galNAc4,6S represented > 96% of the terminal residues. There was a significant decrease in 6-sulfation of the terminal residues in degenerative cartilage.

Conclusions

6-Sulfation of internal and terminal CS residues increased with age. Cartilage degeneration in racehorses was accompanied by deposition of CS chains with altered sulfation patterns. In normal and diseased joints of horses > 2 years old, synovial fluid CS was not indicative of cartilage CS and may represent turnover products of a subpopulation of proteoglycan within the matrix. (Am J Vet Res 1998;59:786-791)

Free access
in American Journal of Veterinary Research

Summary

Five healthy adult mares and 1 gelding were given a single dose (15 mg/kg of body weight) of metronidazole per rectum. After manual evacuation of feces from the rectum, a suspension of crushed tablets and water (40 ml) was administered via a 28-F catheter advanced 30 cm into the rectum. Blood samples were obtained by jugular venipuncture, and metronidazole concentration was measured serially for the 14 hours after drug administration. Mean serum concentration of metronidazole peaked at 4.5 μg/ml, 0.83 hour after administration, and decreased to 0.38 μg/ml, 14 hours after administration. Mean elimination rate constant was 0.23/h, and the harmonic mean elimination half-life was 3.04 hours. Further study is necessary to determine a therapeutic dose regimen for metronidazole administered per rectum.

Free access
in American Journal of Veterinary Research

Abstract

Objective—To determine the pharmacokinetics of azithromycin and its concentration in body fluids and bronchoalveolar lavage cells in foals.

Animals—6 healthy 6- to 10-week-old foals.

Procedure—Azithromycin (10 mg/kg of body weight) was administered to each foal via IV and intragastric (IG) routes in a crossover design. After the first IG dose, 4 additional IG doses were administered at 24-hour intervals. A microbiologic assay was used to measure azithromycin concentrations in serum, peritoneal fluid, synovial fluid, pulmonary epithelial lining fluid (PELF), and bronchoalveolar (BAL) cells.

Results—Azithromycin elimination half-life was 20.3 hours, body clearance was 10.4 ml/min·kg, and apparent volume of distribution at steady state was 18.6 L/kg. After IG administration, time to peak serum concentration was 1.8 hours and bioavailability was 56%. After repeated IG administration, peak serum concentration was 0.63 ± 0.10 µg/ml. Peritoneal and synovial fluid concentrations were similar to serum concentrations. Bronchoalveolar cell and PELF concentrations were 15- to 170-fold and 1- to 16-fold higher than concurrent serum concentrations, respectively. No adverse reactions were detected after repeated IG administration.

Conclusions and Clinical Relevance—On the basis of pharmacokinetic values, minimum inhibitory concentrations of Rhodococcus equi isolates, and drug concentrations in PELF and bronchoalveolar cells, a single daily oral dose of 10 mg/kg may be appropriate for treatment of R equi infections in foals. Persistence of high azithromycin concentrations in PELF and bronchoalveolar cells 48 hours after discontinuation of administration suggests that after 5 daily doses, oral administration at 48-hour intervals may be adequate. (Am J Vet Res 2001;62:1870–1875)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine regional and zonal variation in sulfation patterns of chondroitin sulfate in normal equine corneal stroma.

Sample Population—22 normal eyes from 11 horses.

Procedure—Corneas were collected within 24 hours of death from equine necropsy specimens. After papain-chondroitinase digestion of corneal tissue, disaccharides ΔDi4S and ΔDi6S were quantified by use of capillary zone electrophoresis in the superficial, middle, and deep zones of central and peripheral regions of the cornea.

Results—For the 2 regions combined,ΔDi6S/ΔDi4S values were significantly lower in the deep and middle zones, compared with that of the superficial zone. In the central region, deep and middle zones had significantly lower ΔDi6S/ΔDi4S values than the superficial zone did. In the peripheral region, the deep zone had significantly lower ΔDi6S/ΔDi4S values, compared with superficial and middle zones. In the deep zone, the peripheral region had significantly lower ΔDi6S/ΔDi4S values than the central region did.

Conclusion and Clinical Relevance—Distribution of ΔDi6S/ΔDi4S values follows a gradient across the healthy equine cornea, being smallest in the deep and middle zones of the central region and the deep zone of the peripheral region. Regional and zonal differences in the distribution of stromal ΔDi6S and ΔDi4S may influence the role of glycosaminoglycans in health, disease, and wound repair of the equine cornea. (Am J Vet Res 2002;63:143–147)

Full access
in American Journal of Veterinary Research