Relative amounts of chondroitin sulfate and hyaluronic acid in synovial fluid from normal and osteochondrotic swine joints.

Twenty eight nonlame and ten lame pigs were used to study glycosaminoglycans in synovial fluid from normal and osteochondrotic elbow and stifle joints. The results indicated that porcine synovial fluid contains both hyaluronic acid and chondroitin sulfate and that the chondroitin sulfate to hyaluronic acid ratio is similar (P less than 0.05) between normal and osteochondrotic joints.     

Urea as a measure of dilution of equine synovial fluid

This paper tests the hypothesis that serum and synovial urea concentrations are similar and that urea concentration can be used as an accurate marker for synovial fluid dilution in normal equine joints. Serum and synovial fluid urea concentrations were compared in 42 horses and were equivalent for individual horses (P<0.0001). Mean +/- s.e. serum concentration was 6.1+/-0.552 mmol/l and synovial concentration 6.0+/-0.459 mmol/l. The normal range for synovial urea concentration was determined as 2.5-7.7 mmol/l. The synovial urea concentration from different synovial structures in individual horses were compared and were equivalent (P = 0.002). Known dilutions of synovial fluid with saline were made. The actual and expected synovial urea concentrations were compared and were equivalent (P<0.001). An accurate method of calculating synovial fluid dilution has been determined.     

Evaluation of the role of keratan sulphate as a molecular marker to monitor cartilage metabolism in horses

The role of keratan sulphate (KS) as a metabolic marker of cartilage was evaluated using an in vitro model of equine articular cartilage. Articular cartilage was harvested from clinically healthy 6-month-old foals (n = 3). Chondrocytes were centrifuged and cultured as pellets. Chondrocyte pellets were stimulated by insulin-like growth factor-I alpha (IGF-I alpha) or interleukin-1 alpha (IL-1 alpha) for 2 weeks. The concentrations of sulphated glycosaminoglycans (GAG) and KS in the culture media were measured by a 1,9-dimethyl-methylene blue (DMMB) colorimetric assay and an inhibition enzyme-linked immunosorbent assay using a 1/20/5D4 antibody, respectively. The concentration of GAG was significantly increased both in the media of pellets stimulated by IGF-I alpha and in those stimulated by IL-1 alpha. KS concentration was significantly increased in those stimulated by IL-1 alpha, while no significant change was found in those stimulated by IGF-I alpha. A high correlation between GAG and KS concentrations was found in the media of pellets stimulated by IL-1 alpha (r = 0.84), but not in those stimulated by IGF-I alpha (r = 0.59). The results suggest that the concentration of KS reacting to 1/20/5D4 mirrors the GAG concentration during the stage of cartilage catabolism, but not during the cartilage anabolic stage. The KS concentration in biological fluids could therefore be a useful marker to understand further the cartilage catabolic process. It may also represent some aspects of the cartilage anabolic process.     

Hyaluronic acid concentration in synovial fluid from normal and arthritic joints of horses

A method previously described was used to determine the hyaluronic acid concentration in synovia from normal and arthritic horse joints. The concentration of hyaluronic acid in the synovia from arthritic joints was found to be significantly lower than the concentration in fluid from normal joints.     

Surgical treatment of synovial osteochondromatosis in the middle carpal joint of a pony

A 9‐year‐old Paint pony gelding presented for signs of left carpal swelling of 1–2 weeks' duration. Radiographic, ultrasonographic and arthroscopic evaluation of the left carpus was consistent with synovial osteochondromatosis. This presumptive clinical diagnosis was confirmed histopathologically. Arthroscopic removal of the osteochondral bodies resulted in resolution of the carpal effusion and return to previous athletic activity by 4.5 months post operatively. Arthroscopic removal of osteochondral bodies is the treatment of choice in cases of suspected synovial osteochondromatosis.     

Gelatinase activity in synovial fluid and synovium obtained from healthy and osteoarthritic joints of dogs

OBJECTIVE: To determine matrix metalloproteinase (MMP) activity in synovial fluid (SF) obtained from the joints of dogs with degenerative joint disease (DJD) secondary to various underlying conditions. SAMPLE POPULATION: 35 samples of SF obtained from 18 clinically normal (control) dogs and 34 samples of SF obtained from 17 dogs with DJD; dogs with DJD were from 2 populations (client-owned dogs and research dogs that had DJD secondary to the lysosomal storage disease mucopolysaccharidosis VII). PROCEDURE: MMP activity in samples of SF was semiquantitatively examined by use of gelatin or casein zymography. Western blot analysis was performed by use of antibodies for MMP-2 and MMP-9. In addition, in situ MMP activity was observed in sections of synovial membrane obtained from healthy and osteoarthritic joints. RESULTS: Samples of SF from osteoarthritic joints had higher MMP-2 activity and dramatically increased MMP-9 activity, compared with values for healthy joints. Substrate-overlay analyses indicated minimal gelatin-degrading activity in synoviocytes obtained from control dogs, whereas greater activity was seen in osteoarthritic synoviocytes, with additional activity in the underlying tissue. CONCLUSIONS AND CLINICAL RELEVANCE: Higher MMP-2 activity and dramatic increases in MMP-9 activity were associated with the osteoarthritic state, even though MMP-2 activity was detected in healthy joints. This study expands information on MMP production in SF of osteoarthritic joints in other species and documents the similarity of MMP activity patterns regardless of the cause of DJD.     

Determination of total protein concentration and viscosity of synovial fluid from the tibiotarsal joints of horses.

Viscosity of synovial fluid (SF) from 29 clinically normal horses was determined by use of a rotational cone and plate microviscosimeter. Total protein concentration in the SF of the 29 horses, as measured with a refractometer, was less than 2.5 g/dl. When the Coomassie brilliant blue test was used to determine total protein concentration in SF for 15 horses, the mean value was 1,088 mg/dl. Viscosity values at 60, 30, 12, 6, 3, and 1.5 revolutions/min (rpm) spindle speed were 4.41 +/- 1.54 centipoise (cp), 5.29 +/- 1.94 cp, 6.76 +/- 2.76 cp, 8.52 +/- 4.27 cp. 10.41 +/- 6.30 cp, and 13.07 +/- 9.05 cp, respectively. Synovial fluid viscosity increased with decreasing rpm and shear rate, but the shape of the curve for each horse fitted the asymptotic curve. The rotational cone and plate microviscosimeter was an accurate instrument in measuring SF viscosity at multiple rpm or shear rates in horses. The values obtained on clinically normal horses in this study will serve as a baseline for comparison in the evaluation of horses with joint disease.     

Cytologic analysis of synovial fluid in clinically normal tarsal joints of young camels (Camelus dromedarius)

BACKGROUND: Camels are important in the racing industry and for milk, meat, and hair production in the Middle East. Evaluation of synovial fluid is an important part of the assessment of musculoskeletal injuries in this species. Information in the literature regarding synovial fluid in camels is limited. OBJECTIVES: The objective of this study was to determine the protein and cellular composition of synovial fluid from the tarsal joints of clinically normal, young camels (Camelus dromedarius). METHODS: Thirty clinically healthy, male camels, aged 9 to 12 months, were used in the study. Synovial fluid samples were collected from the right and left tarsal joints. Samples were processed within 60 minutes after collection. Total nucleated cell counts (TNCC) were assessed using a hemacytometer. Total protein concentration was determined using a refractometer. RESULTS: Forty-six samples were analyzed. The TNCC (mean +/- SD) was 175.8 +/- 136.7 cells/microL (range 50-678 cells/microL). Differential cell percentages were obtained for lymphocytes (58.2 +/- 21.55%, range 15-90%), monocyte/macrophages (38.3 +/- 20.8%, range 10-85%), and neutrophils (3.5 +/- 5.1%, range 0-15%). Protein concentration was 2.1 +/- 0.6 g/dL (range 1-3 g/dL). Significant differences were not observed in any parameters between right and left tarsal joints. CONCLUSION: Synovial fluid reference values were established and may be useful in the clinical investigation of joint disease in young camels.     

Polypoid cystitis as a cause of haematuria in a pony mare

A 15-year-old pony mare was presented for investigation of haematuria of 2 weeks' duration. On cystoscopy, multiple small pedunculated soft tissue structures were observed on the bladder mucosa. Histopathological analysis of the masses was consistent with chronic polypoid cystitis. The polypoid lesions and associated haematuria resolved following prolonged antibiotic treatment. Polypoid cystitis has not previously been described in horses. This condition should be considered a differential for haematuria, requiring cystoscopy and biopsy to confirm a diagnosis.     

Assessment of glycosaminoglycan concentration in equine synovial fluid as a marker of joint disease.

A modification of a colorimetric assay was used to determine synovial fluid total and individual sulphated-glycosaminoglycan concentration in various clinical presentations of joint disease in horses. Concentrations of synovial fluid and serum sulphated-glycosaminoglycan (GAG) were measured by the 1,9-dimethylmethylene blue (DMMB) dye assay in normal horses (n = 49), horses with acute (n = 26) or chronic (n = 27) joint disease (defined by clinical, radiographic, and clinicopathological parameters), and horses with cartilaginous lesions at diagnostic arthroscopy, but with normal radiographs and synovial fluid (n = 9). Horses with acute joint disease were subdivided into moderate acute (n = 21) and severe acute (n = 5) joint disease on the basis of synovial fluid analysis and clinical examination. Horses with chronic joint disease were subdivided into mild chronic (n = 9), moderate chronic (n = 10), and severe chronic (n = 8) joint disease on the basis of synovial fluid analysis, clinical examination, and radiographic findings. The concentrations of chondroitin sulphate (CS) and keratan sulphate (KS) were analyzed in each sample following sequential enzymatic digestion of the sample with chondroitinase or keratanase. In addition, the concentration of hyaluronate (HA) in each sample was determined by a colorimetric assay following digestion of the sample with microbial hyaluronidase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Concentrations of substance P and prostaglandin E2 in synovial fluid of normal and abnormal joints of horses

OBJECTIVE: To correlate substance P content of synovial fluid with prostaglandin E2 content, radiographic evidence of joint abnormality, and anatomic location of the joint for normal and osteoarthritic joints of horses. SAMPLE POPULATION: Synovial fluid from 46 normal joints in 21 horses and 16 osteoarthritic joints in 10 horses. PROCEDURE: Normal and osteoarthritic joints were identified by clinical and radiographic examination, by response to nerve blocks, during scintigraphy or surgery, or by clinicopathologic evaluation. Substance P and prostaglandin E2 contents of synovial fluid were determined by radioimmunoassay. Radio-graphs of joints were assigned a numeric score reflecting severity of lesions. Joints were assigned a numeric score reflecting anatomic location. RESULTS: Median concentrations of substance P and prostaglandin E2 were significantly increased in osteoarthritic joints, compared with normal joints. A significant correlation was found between concentrations of substance P and prostaglandin E2 in synovial fluid, but a correlation was not detected between substance P concentration in synovial fluid and anatomic location of the joint or between radiographic scores of osteoarthritic joints and concentrations of substance P or prostaglandin E2. CONCLUSIONS AND CLINICAL RELEVANCE: A correlation existed between concentrations of substance P and prostaglandin E2 in synovial fluid obtained from normal and osteoarthritic joints. However, content of substance P in synovial fluid cannot be predicted by the radiographic appearance of the joint or its anatomic location. Substance P and prostaglandin E2 may share an important and related role in the etiopathogenesis of osteoarthritis, lending credence to the importance of neurogenic inflammation in horses.     

Gentamicin concentrations in synovial fluid obtained from the tarsocrural joints of horses after implantation of gentamicin-impregnated collagen sponges

OBJECTIVE: To determine synovial fluid gentamicin concentrations and evaluate adverse effects on the synovial membrane and articular cartilage of tarsocrural joints after implantation of a gentamicin-impregnated collagen sponge. ANIMALS: 6 healthy adult mares. PROCEDURES: A purified bovine type I collagen sponge impregnated with 130 mg of gentamicin was implanted in the plantarolateral pouch of 1 tarsocrural joint of each horse, with the contralateral joint used as a sham-operated control joint. Gentamicin concentrations in synovial fluid and serum were determined for 120 hours after implantation by use of a fluorescence polarization immunoassay. Synovial membrane and cartilage specimens were collected 120 hours after implantation and evaluated histologically. RESULTS: Median peak synovial fluid gentamicin concentration of 168.9 microg/mL (range, 115.6 to 332 microg/mL) was achieved 3 hours after implantation. Synovial fluid gentamicin concentrations were < 4 microg/mL by 48 hours. Major histologic differences were not observed in the synovial membrane between control joints and joints implanted with gentamicin-impregnated sponges. Safranin-O fast green stain was not reduced in cartilage specimens obtained from treated joints, compared with those from control joints. CONCLUSIONS AND CLINICAL RELEVANCE: Implantation of a gentamicin-impregnated collagen sponge in the tarsocrural joint of horses resulted in rapid release of gentamicin, with peak concentrations > 20 times the minimum inhibitory concentration reported for common pathogens that infect horses. A rapid decrease in synovial fluid gentamicin concentrations was detected. The purified bovine type I collagen sponges did not elicit substantial inflammation in the synovial membrane or cause mechanical trauma to the articular cartilage.     

Bacteroides sp. as a cause of anaerobic keratitis in a pony

This Case Report describes anaerobic infection of the conjunctiva and cornea in a pony. A 13‐year‐old pony was admitted for evaluation following suspected ocular trauma. Bacterial conjunctivitis and keratitis were diagnosed based on clinical appearance of the eye and cytological evaluation of a corneal scraping. The pony responded poorly to topical therapy, and enucleation was performed. Culture of the corneal scraping yielded Corynebacterium sp. and Bacteroides sp. Anaerobic keratitis and conjunctivitis is uncommonly reported in horses, but should be suspected when there is a history of ocular trauma or skin disease, or when a malodorous ocular discharge is identified.     

H-PSGAG concentration in the synovial fluid of the equine antebrachiocarpal, metacarpophalangeal, coronopedal and tibiotarsal joints following a 500 MG i

Eight mature horses were administered a single intramuscular injection of 500 mg polysulfated glycosaminoglycan (PSGAG) labeled with 2.044 mCi tritium. Synovial fluid samples were collected from the antebrachiocarpal (carpal), metacarpophalangeal (fetlock), tibiotarsal (hock) and coronopedal (coffin) joints prior to injection and at 2, 4, 8, 12, and 24 hours after injection. The samples were subjected to scintillation counting in decays per minute and were converted to μg PSGAG per ml. The levels achieved in the synovial fluid of the various joints were compared to levels of PSGAG described as adequate to inhibit enzymes which degrade articular cartilage matrix components and hyaluronic acid and adequate to stimulate production of new matrix components and hyaluronic acid in diseased joints.Mean synovial fluid ³H-PSGAG levels indicated that peak concentrations of ³H-PSGAG were achieved 2 hours post injection in all joints and that these concentrations were within the therapeutic range for PSGAG. The peak concentrations were not significantly different among the joints except between the antebrachiocarpal and the metacarpophalangeal joints. The areas under the concentration-time curves (AUC) for each joint were computed by the trapezoidal method from hour 0 through hour 24 and by empirical exponential decay beyond hour 24. These values were subjected to an analysis of variance (ANOVA). The overall multivariate test of AUC among all joints was not significant.The data from this study indicate that a single intramuscular 500 mg injection of PSGAG provided therapeutic levels of the drug in the equine antebrachiocarpal, metacarpophalangeal, tibiotarsal, and coronopedal joints within 2 hours of injection. While there were differences in levels between joints at certain time points, the AUC values suggest similar distribution of the drug in all joints tested.     

Consideration of the role of antigenic keratan sulphate reacting to a 1/14/16H9 antibody as a molecular marker to monitor cartilage metabolism in horses

The role of keratan sulphate (KS) as a marker of cartilage metabolism was evaluated by using an in vitro model of equine articular cartilage. Articular cartilage was harvested from clinically healthy 6-month-old foals (n=3). Chondrocytes were centrifuged and cultured as pellets. Chondrocyte pellets were stimulated by insulin-like growth factor (IGF)-Ialpha or interleukin (IL)-1alpha for 2 weeks. The sulfated glycosaminoglycans (GAG) and antigenic KS concentrations in the culture media were measured by a 1,9-dimethyl-methylene blue (DMMB) colorimetric assay and an inhibition ELISA using a 1/14/16H9 antibody, respectively. Concentration of GAG was significantly increased in the media of pellets stimulated by both IGF-Ialpha and IL-1alpha. Antigenic KS concentration was significantly increased in those stimulated by IL-1alpha, while no significant change was found in those stimulated by IGF-Ialpha. A high correlation between GAG and antigenic KS concentrations was found in the media of pellets stimulated by IL-1alpha (r=0.87), but not in those stimulated by IGF-Ialpha (r=0.43). The results suggest that the concentration of antigenic KS reacting to 1/14/16H9 mirrors the GAG concentration during the stage of cartilage catabolism, but not during the cartilage anabolic stage. The concentration of antigenic KS reacting to 1/14/16H9 antibody in biological fluids could therefore be a useful marker to further understand principally the catabolic and slightly the anabolic process of articular cartilage metabolism.