Expression of immune response genes in the stifle joint of dogs with oligoarthritis and degenerative cranial cruciate ligament rupture

Dysregulation of immune responses within joints plays an important role in development of inflammatory arthritis. We determined expression of a panel of immune response and matrix turnover genes in synovial fluid collected from a group of dogs with stifle oligoarthritis and associated degenerative cranial cruciate ligament (CCL) rupture (n=27). We also studied synovial fluid gene expression in dogs affected with other forms of degenerative arthritis (n=9) and in the stifle joint of healthy dogs with intact CCL (n=14). After collection, synovial cells were pelleted and RNA was isolated. Relative expression of cathepsin K, cathepsin S, tartrate-resistant acid phosphatase (TRAP), matrix metalloproteinase-9 (MMP-9), invariant chain (li), toll-like receptor-2 (TLR-2), and TLR-9 was determined using real-time quantitative RT-PCR. Data were normalized to peripheral blood mononuclear cells (PBMC) as an internal control. Relative expression of cathepsin K, MMP-9, TRAP, and li was increased in the stifle synovial fluid of dogs with oligoarthritis, when compared with the stifles of healthy dogs (P<0.05). In contrast, relative expression of all of the genes-of-interest in synovial fluid from joints affected with other forms of arthritis was not significantly different from the stifles of healthy dogs. TRAP expression was also significantly increased in the stifle joints of dogs with oligoarthritis, when compared to joint expression of TRAP in dogs with other forms of degenerative arthritis (P<0.05). In the dogs with stifle oligoarthritis, expression of both matrix turnover and immune response genes was increased in stifle synovial fluid, when compared with the internal PBMC control, whereas in healthy dogs and dogs with other forms of arthritis, only expression of matrix turnover genes was increased in synovial fluid, when compared with the internal PBMC control (P<0.05). Taken together, these findings suggest that antigen-specific immune responses within the stifle joint may be involved in the pathogenesis of persistent synovitis and associated joint degradation in dogs with oligoarthritis and degenerative CCL rupture.     

Absolute and relative cell counts for synovial fluid from clinically normal shoulder and stifle joints in cats

OBJECTIVE: To determine absolute and relative cell counts for synovial fluid from grossly, radiographically, and histologically normal shoulder and stifle joints in healthy cats. DESIGN: Clinical study. ANIMALS: 52 cats scheduled to be euthanatized for unrelated reasons. PROCEDURE: Arthrocentesis of the shoulder and stifle joints was performed bilaterally, and synovial fluid was analyzed for absolute WBC count, WBC morphology, and percentages of neutrophils and mononuclear cells. Joints were examined grossly and radiographically, and synovial membrane specimens were submitted for histologic examination. Synovial fluid samples that were contaminated with blood and samples from joints with any gross, radiographic, or histologic abnormalities were excluded. RESULTS: 82 of the 208 synovial fluid samples were excluded because abnormalities were identified during physical examination; the volume of fluid obtained was insufficient for analysis; there was evidence of blood contamination; or the joint had gross, radiographic, or histologic abnormalities. Median WBC count for the remaining 126 synovial fluid samples was 91 cells/microL (96.4% mononuclear cells and 3.6% neutrophils); WBC count was not significantly different between left and right joint samples or between shoulder and stifle joint samples. Body weight was associated with synovial fluid WBC count, with WBC count increasing as body weight increased. Sixteen of the 52 (30%) cats had radiographic evidence of osteoarthritis involving at least 1 joint. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that synovial fluid can be obtained reliably from shoulder and stifle joints in cats.     

Correlation between osteoarthritic changes in the stifle joint in dogs and the results of orthopedic,radiographic, ultrasonographic and arthroscopic examinations

Osteoarthritis (OA) is a chronic, degenerative disease affecting the articular cartilage and subchondral bone that causes pain and inhibits movement. The stifle’s joint fibrous capsule contains the synovial membrane, which produces cartilage nutrients. A ruptured cranial cruciate ligament injures the joint and produces OA. Osteoarthritis diagnosis starts with clinical radiographic and ultrasonographic tests, although the latter is not used very much in dog and cat clinics for this purpose. The objective of this study was to establish the correlation among the results of orthopedic, radiographic, ultrasonographic examinations and structural anatomical changes revealed by arthroscopic evaluation to diagnose stifle joint OA and determine risk factors in the dogs affected. Of 44 clinical cases of OA included in the study, 88.64% had ruptured of cranial cruciate ligaments. The correlation between synovial fluid effusion and osteophytosis was of 0.84. It was concluded that there is good diagnostic agreement between synovial fluid effusion and osteophytosis when dealing with stifle joint OA. Risk factors for dogs regarding the development of stifle joint OA included: ruptured cranial cruciate ligaments or patella luxation, female dogs and weight over 10 kg.     

Serum and synovial fluid concentrations of keratan sulfate and hyaluronan in dogs with induced stifle joint osteoarthritis following cranial cruciate ligament transection

OBJECTIVE: To examine longitudinal changes in serum and synovial fluid concentrations of keratan sulfate (KS) and hyaluronan (HA) after cranial cruciate ligament (CCL) transection in dogs. ANIMALS: 12 clinically normal adult mixed-breed dogs. PROCEDURE: Following CCL transection in the right stifle joint, KS and HA concentrations were determined in serum and neat (undiluted) synovial fluid prior to and 1, 2, 3, and 12 months after surgery. Postsurgical dilution of synovial fluid was corrected by use of urea as a passive marker. RESULTS: Synovial fluid KS and HA concentrations decreased at 1, 2, and 3 months after surgery in operated stifle joints, compared with baseline values. Synovial fluid KS concentration decreased in unoperated stifle joints at 1 month. A decrease in synovial fluid KS concentration was found in operated stifle joints, compared with unoperated stifle joints, at 2 and 3 months, and a decrease in synovial fluid HA concentrations was also found in operated stifle joints, compared with unoperated stifle joints, at 1, 2, and 3 months. Serum KS concentrations increased from baseline values at 3 months after surgery. Hyaluronan concentrations in operated stifle joints were lower than baseline values at 1, 2, and 3 months. Urea-adjusted synovial fluid concentrations revealed that dilution did not account for the decline in biomarker concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: The initial decrease and subsequent increase in synovial fluid concentrations of HA and KS may be caused by an acute inflammatory response to surgical intervention that negatively affects cartilage metabolism or an increase in production of immature proteoglycans.     

LOW-FIELD MAGNETIC RESONANCE IMAGING OF THE CANINE STIFLE JOINT: NORMAL ANATOMY

Low-field magnetic resonance imaging (MRI) was performed on the stifle joints of four normal adult mongrel dogs using a 0.064 Tesla scanner. Markers were placed on each stifle joint to serve as reference points for comparing gross sections with the images. A T1-weighted sequence was used to image one stifle joint on each dog in the sagittal plane and the other stifle joint in the dorsal plane. The dogs were euthanized immediately following MRI and the stifle joints frozen intact. Each stifle joint was then embedded in paraffin, again frozen, and sectioned using the markers as reference points. On T1-weighted images, synovial fluid had low signal intensity (dark) compared to the infrapatellar fat pad which had a high signal intensity (bright). Articular cartilage was visualized as an intermediate bright signal and was separated from trabecular bone by a dark line representing subchondral bone. Menisci, fibrous joint capsule, and ligamentous structures appeared dark. In the true sagittal plane, the entire caudal cruciate ligament was often seen within one image slice. The patella was visualized as an intermediate bright signal (trabecular bone) surrounded by a low intensity signal (cortical bone). The trochlea and the intercondylar notch were difficult areas to analyze due to signal volume averaging of the curved surface of these areas and the presence of several types of tissues.     

Expression of interleukin-8 and intercellular cell adhesion molecule-1 in the synovial membrane and cranial cruciate ligament of dogs after rupture of the ligament

This cross-sectional clinical study compared inflammation, including expression of the chemokine interleukin (IL)-8 and intercellular cell adhesion molecule-1 (ICAM-1), in the stifle joints of 4 control dogs and 23 dogs with cranial cruciate ligament rupture (CCLR). The CCL, synovial membrane, meniscus, cartilage, and synovial fluid from the affected stifle joints of all the dogs were examined. Inflammatory cell counts were performed on the synovial fluid, and the tissues were processed for histologic study and immunohistochemical detection of IL-8 and ICAM-1. The synovial fluid from the stifle joints of the dogs with CCLR had an increased percentage of neutrophils (P = 0.054) and a decreased percentage of lymphocytes (P = 0.004) but not macrophages compared with the fluid from the control dogs. There was accumulation of inflammatory cells and increased expression of IL-8 and ICAM-1 in the vascular endothelium of the synovial membrane and the CCL of the dogs with CCLR. The increase in inflammatory cells in the stifle joints of dogs with CCLR may therefore be due to increased expression of IL-8 and ICAM-1 in the synovial membrane and the CCL after the injury. These data may help in understanding the mechanisms of inflammation associated with CCLR.     

Evaluation of anticollagen type I antibody titers in synovial fluid of both stifle joints and the left shoulder joint of dogs with unilateral cranial cruciate disease

OBJECTIVE: To evaluate anticollagen type I antibodies in synovial fluid of the affected stifle joint, the contralateral stifle joint, and the left shoulder joint of dogs with unilateral cranial cruciate ligament (CrCL) rupture during an extended period of 12 to 18 months. ANIMALS: 13 client-owned dogs with CrCL rupture and 2 sham-operated dogs. PROCEDURES: All dogs were examined and arthrocentesis of all 3 joints was performed every 6 months after surgery. Synovial fluid samples were tested for anticollagen type I antibodies by use of an ELISA. RESULTS: Dogs with partial CrCL rupture had higher antibody titers than dogs with complete rupture. Six of 13 dogs ruptured the contralateral CrCL during the study, whereby higher antibody titers were found for the stifle joints than for the shoulder joint. Seronegative dogs or dogs with extremely low antibody titers and 2 dogs with high antibody titers did not sustain a CrCL rupture in the contralateral stifle joint. CONCLUSIONS AND CLINICAL RELEVANCE: In most dogs that had a CrCL rupture of the contralateral stifle joint, a distinct antibody titer gradient toward the stifle joints was detected, suggesting that there was a local inflammatory process in these joints. However, only a small number of sham-operated dogs were used to calculate the cutoff values used to determine the anticollagen type I antibody titers in these patients. Synovial fluid antibodies against collagen type I alone do not initiate CrCL rupture because not all dogs with high antibody titers sustained a CrCL rupture in the contralateral stifle joint.     

Anatomical and functional communications between the synovial sacs of the equine stifle joint

The anatomical and functional communications of the synovial sacs of the equine stifle joint were evaluated in 50 stifle joints of 25 horses. Femoropatellar joint (FPJ) sacs were injected with 50 ml of gelatin-based dye and horses were then walked for 50 m. Horses were subsequently killed, the stifle joints dissected and the location of the dye recorded. Twenty-three horses (46 joints) had clinically normal stifle joints and in this group, anatomical communications of the stifle joints were bilaterally symmetrical in each horse. In 15 of these 23 horses (65 per cent), direct anatomical communication between the FPJ sac and the medial sac of the femorotibial joint (FTJ) was demonstrated. The FPJ sac communicated with both the medial and lateral sacs of the FTJ in four of these 23 horses (17.5 per cent). There were no anatomical communications between the FPJ sac and either sac of the FTJ in the remaining four horses (17.5 per cent). Functional communication, which was established by finding dye in the FTJ sacs were anatomical communication with the FPJ sac existed, was demonstrated in 14 of 19 horses (74 per cent). Two horses were affected with degenerative joint disease of one stifle joint. In both of these joints the FPJ sac communicated with both the medial and lateral FTJ sacs. This distribution was different from that of the contralateral joint. When performing intra-articular anaesthesia of equine stifle joints, each synovial sac needs to be injected separately to ensure that anaesthesia of the appropriate synovial sac is obtained.     

Evaluation of Experimental Transection and Partial Excision of the Caudal Cruciate Ligament in Dogs

The caudal cruciate ligament (CaCL) of one stifle joint in seven dogs was transected and a 2 to 4 mm section was removed. Six months after surgery, none of the dogs were lame. Thigh muscle circumference, stifle range of motion, and internal tibial rotation in the operated limb were not significantly different from the preoperative measurements or the contralateral, unoperated limb. A caudal drawer motion was consistently present in the stifle joints with a transected CaCL. A radiographic evaluation of the operated stifle joints did not reveal osteoarthritic changes; four of seven stifle joints had an irregular fat pad 6 months after surgery. Results of a joint fluid analysis revealed a slight increase in synovial cells within treated stifle joints; inflammatory cells were not observed. The only gross morphologic change in stifle joints with a severed ligament was enlarged knobby remnants of the CaCL. Articular cartilage defects or osteophytes were not observed. Results of a histologic examination of the CaCL remnants revealed synovial cellular capping and intraligamentous fibroplasia. Based on a limited number of dogs, it was concluded that isolated transection of the CaCL produced minimal clinical and pathologic changes in the stifle joint during a 6 month period.     

Diffusion of mepivacaine between adjacent synovial structures in the horse. Part 2: tarsus and stifle

This paper tests the hypothesis that the local analgesic agent mepivacaine diffuses between adjacent equine synovial structures in the hindlimb and with greater frequency than latex, gelatine dye or contrast media. We report the incidence of diffusion of mepivacaine between the tarsometatarsal, centrodistal and tarsocrural joints, and the 3 synovial compartments of the stifle in 33 fresh equine cadavers. The tarsometatarsal joint and one synovial compartment of the stifle in the left limb and the centrodistal joint and a different synovial compartment of the stifle in the right limbs were injected with mepivacaine. Following flexion and extension of the limb, synovial fluid was aspirated from the noninjected centrodistal and tarsometatarsal joints and the tarsocrural joints of the hock and the noninjected compartments of the stifle. Concentrations of mepivacaine in these samples were assayed using an enzyme linked immunosorbent assay. For samples obtained by dilution of synovial fluid the concentration of mepivacaine was determined by comparing the concentration of urea in the diluted synovial fluid and the concentrations of the serum urea. Mepivacaine was detected in 25/25 (100%) adjacent tarsometatarsal and centrodistal joints after diffusion in both directions, in 23/25 (92%) of tarsocrural joints after diffusion from tarsometatarsal joints and in 22/25 (88%) tarsocrural joints after diffusion from centrodistal joints in the hocks. Diffusion from the femoropatellar to medial and lateral femorotibial joints and between the medial and lateral femorotibial joints in both directions were 20/20 (100%). Diffusion from the lateral femorotibial to the femoropatellar joint was 18/20 (90%) and from the medial femorotibial to femoropatellar joints 17/20 (85%). Mepivacaine was detected at concentrations >0.3 mg/l in a proportion of samples ranging from 15/25 (60%) in the tarsocrural joint following tarsometatarsal joint injection to 18/20 (90%) in the lateral femorotibial joint after femoropatellar joint injection. At mepivacaine concentrations >100 mg/l, detection ranged from 3/20 (15%) in the lateral femorotibial joint from the medial femorotibial joint to 19/25 (76%) in the centrodistal joint from the tarsometatarsal joint. At mepivacaine concentrations >300 mg/l, detection ranged from 1/25 (4%) in the tarsocrural joint from the tarsometatarsal joint to 16/25 (64%) in the from centrodistal joint the tarsometatarsal joint. The results show greater diffusion of mepivacaine between these adjacent synovial structures than assumed from previous anatomical, latex injection and contrast arthrographic studies. Therefore, commonly performed intrasynovial local analgesic techniques in the hindlimb of the horse are not as specific as first thought.     

Prevalence and relevance of antibodies to type-I and -II collagen in synovial fluid of dogs with cranial cruciate ligament damage

OBJECTIVE: To measure and compare synovial fluid antibody titers to type-I and -II collagen in stifle joints with instability caused by complete or partial cranial cruciate ligament (CCL) rupture and joints with osteoarthrosis secondary to other pathologic changes in dogs. ANIMALS: 82 dogs with diseased stifle joints. PROCEDURE: Synovial fluid samples were collected from 7 dogs with clinically normal stifles (control group) and 82 dogs with diseased joints (50 stifle joints with complete rupture of the CCL, 20 with partial damage of the CCL, and 12 joints with radiographic signs of osteoarthritis secondary to other arthropathies). Synovial fluid samples were tested for autoantibodies to type-I and -II collagen by an ELISA. RESULTS: In dogs with complete and partial CCL rupture, synovial fluid antibody titers to type-I and -II collagen were significantly increased, compared with control dogs. Forty-eight percent (24/50) of samples from dogs with complete CCL rupture and 35% (7/20) of samples from dogs with partial CCL rupture had antibody titers to type-I collagen that were greater than the mean plus 2 standard deviations of the control group titers. Synovial fluid antibody titers to type-II collagen were high in 40% of the dogs with partial or (8/20) complete (20/50) CCL rupture. Dogs with osteoarthrosis secondary to other pathologic changes had significantly increased synovial fluid antibodies to type-I and -II collagen, compared with control dogs. CONCLUSION: Increases in autoantibodies to collagen in synovial fluid are not specific for the type of joint disorder. It is unlikely that the anticollagen antibodies play an active role in the initiation of weakening of the CCL.     

Multiple joint metastasis of a transitional cell carcinoma in a dog

An 8‐year‐old castrated male hound mix was referred to the Purdue University Veterinary Teaching Hospital for severe lameness, pollakiuria, and dyschezia. On presentation, the dog was nonweight bearing on the right rear limb and the right carpus was diffusely swollen. Synovial fluid analysis from the right carpus revealed a population of epithelial cells displaying marked anisocytosis, anisokaryosis, multinucleation, and prominent, variably sized nucleoli. A metastatic carcinoma with presumed prostatic or urothelial origin was diagnosed based on cytomorphology. Subsequent cytologic evaluation of peripheral lymph nodes revealed the presence of a similar neoplastic population. The dog was euthanized and synovial fluid from both stifle joints, as well as impression smears of the prostate gland, were collected. Carcinoma cells were identified in each stifle joint and in the prostate gland. Immunocytochemistry was performed on synovial fluid smears from 2 of the joints (right stifle and right carpus) and on impression smears of the prostate gland. The neoplastic population in the joints and prostate gland showed strong immunoreactivity to uroplakin III, a urothelial marker, indicating metastasis of a transitional cell carcinoma to multiple joints. In addition, evidence for epithelial to mesenchymal transition was identified using cytokeratin, an epithelial marker, and vimentin, a mesenchymal marker. A necropsy was performed and histopathology confirmed the presence of metastatic transitional cell carcinoma in various tissues. This case illustrates the importance of considering metastatic disease when a patient is presented with severe lameness and joint pain, and the clinical utility of synovial fluid cytology for diagnosis of metastasis in these cases.     

In vivo effects of carprofen, deracoxib, and etodolac on prostanoid production in blood, gastric mucosa, and synovial fluid in dogs with chronic osteoarthritis

OBJECTIVE: To evaluate in vivo activity of carprofen, deracoxib, and etodolac on prostanoid production in several target tissues in dogs with chronic osteoarthritis. ANIMALS: 8 dogs with chronic unilateral osteoarthritis of the stifle joint. PROCEDURE: Each dog received carprofen, deracoxib, or etodolac for 10 days with a 30- to 60-day washout period between treatments. On days 0, 3, and 10, prostaglandin (PG) E2 concentrations were measured in lipopolysaccharide-stimulated blood, synovial fluid, and gastric mucosal biopsy specimens; PGE1 concentrations were measured in gastric mucosal biopsy specimens; and thromboxane B2 (TXB2) was evaluated in blood. RESULTS: Carprofen and deracoxib significantly suppressed PGE2 concentrations in blood at days 3 and 10, compared with baseline, whereas etodolac did not. None of the drugs significantly suppressed TXB2 concentrations in blood or gastric PGE1 synthesis at any time point. All 3 drugs significantly decreased gastric synthesis of PGE2 at day 3 but not day 10 of each treatment period. All 3 drugs decreased synovial fluid PGE2 concentrations in the affected and unaffected stifle joints at days 3 and 10. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that carprofen and deracoxib act in vivo on target tissues as COX-1-sparing drugs by sparing gastric PGE1 and PGE2 synthesis and production of TXB2 by platelets. Etodolac also appears to be COX-1 sparing but may have variable effects on COX-2 depending on the tissue. In gastric mucosa and synovial fluid, there were no significant differences in PG production between compounds at recommended concentrations.     

Effect of Repeated Arthrocentesis on Cytologic Analysis of Synovial Fluid in Dogs

Background: Serial arthrocentesis and synovial fluid examination can be used to monitor treatment efficacy in immune-mediated polyarthritis (IMPA), but whether this procedure induces inflammation that interferes with test result interpretation is unknown.
Objectives: The aim of this study was to determine the effect of repeated arthrocentesis on synovial fluid cytology in healthy dogs.
Animals: Nine healthy client-owned dogs.
Methods: Prospective study. Arthrocentesis was performed under sedation on 4 joints (both carpi, 1 tarsus, 1 stifle) on each dog every 3 weeks, a total of 4 times. Automated cell counts were done on stifle fluid, smears were made, and differential cell counts done on smears from all joints. Slides were evaluated microscopically for erythrocyte numbers, total nucleated cell count, differential cell count, and cell morphology. Data were analyzed by 2-way analysis of variance.
Results: A total of 144 synovial fluid samples were examined. Repeated arthrocentesis was not associated with increases in synovial fluid neutrophil numbers. Mild mononuclear inflammation was detected in 13 samples from 6 dogs.
Conclusions and Clinical Importance: Serial arthrocentesis at 3-week intervals can rarely be associated with mild mononuclear joint inflammation, but does not appear to induce neutrophilic inflammation, at least in healthy dogs, and can be useful to monitor treatment response in canine IMPA.     

Proinflammatory cytokine activities, matrix metalloproteinase-3 activity, and sulfated glycosaminoglycan content in synovial fluid of dogs with naturally acquired cranial cruciate ligament rupture

OBJECTIVE: To measure and compare activities of interleukin-1beta (IL-1beta), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and matrix metalloproteinase-3 (MMP-3); as well as sulfated glycosaminoglycan (S-GAG) content in synovial fluid from dogs with cranial cruciate ligament rupture (CCLR) and dogs with clinically normal stifles. To determine whether correlations exist between demographic and disease-related variables and these synovial markers. STUDY DESIGN: Prospective clinical study. ANIMALS: Dogs with CCLR (n=23) and Beagles with normal stifle joints (n=21). METHODS: Synovial fluid activities of proinflammatory cytokines (IL-1beta, IL-6, and TNF-alpha) were determined by bioassay. MMP-3 activity was measured using fluorogenic substrate. S-GAG contents were determined by dimethylmethylene blue dye-binding assay. Mann-Whitney U-test was used to compare results from CCLR joints with normal controls. Spearman's rank correlation test was used to evaluate associations between demographic and disease-related markers and synovial markers. RESULTS: Mean values for synovial markers were significantly higher in CCLR joints compared with controls. IL-1beta and MMP-3 were positively correlated with lameness duration. CONCLUSIONS: Activities of proinflammatory cytokines, MMP-3 activity and S-GAG contents were significantly elevated in synovial fluid from canine stifle joints with naturally acquired CCLR. These results indicate that there is joint inflammation and increased release of GAGs into synovial fluid, suggesting that these inflammatory changes are associated with depletion of proteoglycan from articular cartilage. CLINICAL RELEVANCE: Medical and surgical treatments designed to decrease joint inflammation and breakdown of proteoglycans may be of value in the management of CCLR in the dog.     

Use of vitamin B12 in joint lavage for determination of dilution factors of canine synovial fluid

OBJECTIVE: To test a modified saline (0.9% NaCl) solution joint washing (lavage) technique that includes the use of vitamin B12 as an internal marker for the evaluation of synovial fluid dilution in lavage samples from canine joints. SAMPLE POPULATION: 9 plasma samples obtained from blood samples of 9 healthy dogs and 9 synovial fluid samples aspirated from stifle joints of 9 cadaveric dogs. PROCEDURE: Photometric absorbances of 25% vitamin B12 solution, canine synovial fluid, and canine plasma were measured in a spectrophotometer to establish an optimal wavelength for analysis. Canine synovial fluid and plasma samples were mixed with the 25% vitamin B12 solution to obtain 1%, 3%, 5%, 10%, 20%, and 50% solutions of synovial fluid or plasma. Diluted synovial fluid and plasma samples were used to simulate joint lavage samples and to examine the possible interference of these substances (synovial fluid or plasma) with the absorbance of the 25% vitamin B12 solution in photometric analysis. RESULTS: The optimal wavelength was found to be at 550 nm. Canine synovial fluid and plasma samples did not interfere with the absorbance measurements of the 25% vitamin B12 solution up to a 50% dilution of plasma or synovial fluid. CONCLUSIONS AND CLINICAL RELEVANCE: The modified saline solution joint lavage method with the use of a 25% vitamin B12 solution as an internal standard provides an accurate and reliable technique for the evaluation of synovial fluid dilution in lavage samples from canine joints.     

Nitric oxide synthase activity in healthy and interleukin 1beta-exposed equine synovial membrane.

OBJECTIVE: To quantitate nitric oxide synthase (NOS) activity in healthy and interleukin 1beta (IL-1beta)-exposed equine synovial membrane. ANIMALS: 6 healthy horses, 2 to 8 years old. PROCEDURE: Recombinant human IL-1beta (0.35 ng/kg of body weight) was injected intra-articularly into 1 metacarpophalangeal joint of each horse. The contralateral joint served as an unexposed control. All horses were euthanatized 6 hours after injection of IL-1beta, and synovial membrane specimens were assayed for NOS activity by measuring conversion of arginine to citrulline. Severity of inflammation was semiquantitated by analysis of synovial fluids and histologic examination of synovial membrane. RESULTS: Equine synovial membrane had minimal NOS activity. A significant difference was not detected in NOS activity between control and IL-1beta-exposed specimens. Histologic examination revealed a neutrophilic infiltrate in synovial membrane exposed to IL-1beta. Synovial fluid from IL-1beta-exposed joints had a moderate inflammatory response and significantly greater concentrations of IL-1beta and interleukin-6 than fluid from healthy joints. CONCLUSION: Healthy equine synovial membrane had low NOS activity that was not affected by exposure to IL-1beta.     

Intra-articular tissue response to analytical grade metrizamide in dogs

The effect of analytical grade metrizamide (AM) injected into the canine stifle, for purposes of arthrography, was studied in 12 adult dogs using saline solution as the control solution injected into the contralateral joints. The AM was used at a concentration of 280 mg of iodine/ml and was administered at the dose of 0.3 ml/cm thickness of the stifle joint. For each joint, arthrocentesis was done before and 7 days after injection of either the contrast medium or saline solution. Physical, biochemical, and cytologic examinations were done on the synovial fluid while the synovial membranes and femoral articular cartilages were sectioned, stained, and examined for histopathologic changes. At the 95% confidence level, significant differences in the total and differential mononuclear cell counts were not seen between the AM- and saline solution-injected joints. However, some subtle changes in the synovial membranes were observed. Intra-articular injection of AM or saline solution initiated a mild inflammatory response, the AM causing slightly more response than the saline solution.