Evaluation of the effects of intra-articular injection of dimethylsulfoxide on normal equine articular tissues

To evaluate the effects of intra-articular injection of dimethylsulfoxide (DMSO) on normal equine articular structures, 7 adult horses with clinically normal carpi were allotted to 2 treatment groups (group A, n = 4; group B, n = 3). In each horse after collection of synovial fluid samples, the right antebrachial carpal and middle carpal joints were aseptically injected with 2 ml of a 40% solution of 90% medical grade DMSO in lactated Ringer solution, and the corresponding joints of the left forelimb (controls) were injected with 2 ml of lactated Ringer solution. In group-A horses, 2 ml of synovial fluid was obtained prior to injections of 40% DMSO at 24 hours and 72 hours, for a total of 3 injections. At necropsy, synovial fluid, synovial membrane, and articular cartilage specimens were obtained. Group-B horses were injected with 40% DMSO in the same sequence; however, the series was repeated following a 1-week interval. Clinical evaluation of these horses revealed no evidence of carpal inflammation associated with any injection in any group. Synovial fluid analysis of DMSO-injected and control joints revealed insignificant differences in leukocyte counts and total protein content. There was no evidence of cartilage degradation on gross, histologic, or histochemical evaluation of any of the joints. Intercellular matrix staining of the articular cartilage failed to reveal any observable difference in glycosaminoglycan content between injection with DMSO or lactated Ringer solution.     

Evaluation of use of dimethyl sulfoxide for intra-articular lavage in clinically normal horses

The antebrachiocarpal and tarsocrural joints of 10 adult horses were randomly assigned to 1 of 4 groups. Groups were formulated and were treated as follows: group 1, control (arthrocentesis only); group 2, buffered lactated Ringer solution; group 3, 10% dimethyl sulfoxide (DMSO; w/v) in lactated Ringer solution; and group 4, 30% DMSO (w/v) in lactated Ringer solution. Joints were lavaged once with the respective solution. Prior to lavage and on days 1, 4, and 8 after lavage, all horses were evaluated for lameness and joint effusion; synovial fluid total and differential WBC counts, synovial fluid total protein concentration, and mucin clot quality were determined. Horses were euthanatized on day 8, and joints were evaluated grossly, histologically, and histochemically. Significant difference was not observed in effect of lactated Ringer solution, 10% DMSO, and 30% DMSO on any measured variable. At 24 hours after treatment, significant (P less than 0.05) difference in synovial fluid WBC numbers and total protein concentration was detected between control and treated joints. Eighty percent of lavaged joints had effusion 24 hours after treatment, compared with 30% of control joints. Gross, histopathologic, or histochemical differences were not detected between treated and control joints. Results of the study indicate that buffered lactated Ringer, 10% DMSO, and 30% DMSO solutions induce similar inflammatory changes in articular structures and significantly greater inflammatory reaction than does arthrocentesis alone.     

Synovial Fluid and Clinical Changes After Arthroscopic Partial Synovectomy of the Equine Middle Carpal Joint

Changes in synovial fluid and clinical variables after arthroscopic partial synovectomy of the middle carpal joint were studied in 12 normal horses. A 7 mm motorized synovial resector was inserted into each middle carpal joint; one middle carpal joint of each horse was randomly selected to have arthroscopic synovectomy (treated) and the opposite joint was lavaged (control). Lameness examinations and synovial fluid analyses were performed before operation and at 8, 14, 21, and 28 days after operation. Lameness variables did not differ between treated and control legs. Middle carpal and carpometacarpal joint circumference measurements were increased for 4 weeks. Synovial fluid specific gravity, pH, total protein, albumin concentration, and alpha-1-, beta- and gamma-globulin concentrations, at 8 and 14 days were significantly higher than before operation in both treated and control middle carpal joints. No significant differences were found between treated and control middle carpal joints at any time for color, clarity, pH, mucin clot formation, total protein, albumin, and globulin fractions. Arthroscopic partial synovectomy and lavage did not cause significant lameness and resulted in a synovitis indistinguishable from synovitis related to arthroscopic lavage alone.     

The effect of implanting gentamicin-impregnated polymethylmethacrylate beads in the tarsocrural joint of the horse

OBJECTIVE: To determine the effect of intra-articular gentamicin-impregnated polymethylmethacrylate (PMMA) beads inserted in the equine tarsocrural joint on the synovial fluid, synovial lining, and cartilage, and to determine the peak and sustainable gentamicin concentrations in synovial fluid and plasma. STUDY DESIGN: Pharmacokinetic, cytologic, and histologic study of the effect of gentamicin-impregnated PMMA on normal equine tarsocrural joints. ANIMALS: Five healthy adult horses. METHODS: Gentamicin-impregnated PMMA bead strands (3 strands each of 40 beads, with each strand containing 100 mg gentamicin) were surgically inserted into one radiographically normal tarsocrural joint in 5 horses. Each horse had both joints flushed with 1 L of lactated Ringer's solution before bead administration. Synovial fluid total protein concentration, white blood cell (WBC) count, gentamicin concentration, synovial histology, cartilage integrity, and cartilage glycosaminoglycan (GAG) concentrations were determined. RESULTS: Gentamicin concentration (mean +/- SEM peak concentration, 27.9 +/- 2.27 microg/mL) occurred in the first 24 hours and remained above 2 microg/mL for 9 days. Gentamicin concentrations in control joints and the plasma remained below detectable levels. The synovial fluid WBC count for treated joints was increased compared with control joints for 72 hours, but was similar at day 6. The synovial protein concentration in gentamicin-treated joints remained increased for 21 days. Synovium in treated joints had diffuse synovitis, whereas control joints had less fibrovascular proliferation. Superficial cartilage erosion was present in all treated joints. There was no difference in the GAG content of treated and control joint cartilage. CONCLUSIONS: Short-term implantation of gentamicin (300 mg)-impregnated PMMA beads can provide therapeutic levels of gentamicin (>2 microg/mL) in the normal tarsocrural joint for 9 days; however, gentamicin-impregnated PMMA beads induce synovitis and superficial cartilage erosion. CLINICAL RELEVANCE: Temporary intra-articular administration of antibiotic-impregnated PMMA may be an effective way to treat septic joints that require constant high concentrations of antibiotics.     

Effects of 0.05% Chlorhexidine Lavage on the Tarsocrural Joints of Horses

In six horses, a 0.05% solution of chlorhexidine diacetate was used to lavage one tarsocrural joint; the contralateral control joint was lavaged with lactated Ringer's solution. Horses were evaluated daily for lameness. Synovial fluid samples were collected on days 1, 4, and 8 for determination of protein concentration, total and differential leukocyte counts, and mucin clot formation. After death on day 8, synovium and osteochondral samples were collected from the tarsocrural joints for examination of morphology and proteoglycan staining. Lavage with chlorhexidine solution caused lameness that was reduced but still evident at day 8. Synovial protein concentration was significantly increased by chlorhexidine lavage; the greatest increase occurred on day 1. Joint lavage increased synovial leukocyte counts on day 1, primarily by increasing polymorphonuclear (PMN) cell counts. Although total synovial leukocyte counts returned to normal by day 4, PMN cell counts remained elevated through day 8; PMN cell counts for chlorhexidine-lavaged joints were typically twice that of control joints. Chlorhexidine lavage caused synovial ulceration, inflammation, and abundant fibrin accumulation. Consistent differences in proteoglycan staining were not detected between control and chlorhexidine-lavaged joints. Joint lavage with 0.05% chlorhexidine diacetate, the lowest known bactericidal concentration, is not recommended for equine joints.     

Evaluation of samarium-153 for synovectomy in an osteochondral fragment-induced model of synovitis in horses

OBJECTIVE: To determine the effects of intraarticular administration of Samarium-153 (153Sm) bound to hydroxyapatite microspheres (153SmM) on an osteochondral chip-induced synovitis. STUDY DESIGN: Sixty days after implantation of autogenous osteochondral fragments in the middle carpal and metacarpophalangeal joints, 153SmM was administered into 1 joint of each type. The contralateral joints were used as untreated controls. ANIMALS OR SAMPLE POPULATION: Fifteen horses without preexisting joint disease were randomly divided into 2 groups (7 in the carpal group, 8 in the metacarpophalangeal group). METHODS: Horses had osteochondral fragments that were harvested from the lateral ridge of the trochlea of the talus and implanted bilaterally into a middle carpal joint and a metacarpophalangeal joint; the opposite joint type served as a control. Sixty days later, 10 to 15 mCi of 153SmM (20 to 50 microm diam) was injected into the fragment-implanted joints. Three horses were treated with nonradioactive hydroxyapatite fragments. Horses were examined clinically until they were killed 14 or 30 days later. Control and treated joints were examined grossly and microscopically to determine the effects of 153SmM on synovial membrane and cartilage. RESULTS: Intraarticular 153SmM caused a transient flare with lameness, effusion, and edema for 48 to 72 hours. Implanted osteochondral chips induced a synovitis characterized by variable degrees of joint damage and synovial infiltrate. Use of 153SmM resulted in synovectomy of variable depth and extent. CONCLUSIONS: Intraarticular 153SmM may be a useful method for synovectomy of inflamed synovial membrane. CLINICAL RELEVANCE: With further testing, radioactive pharmaceuticals might become useful clinical treatments for persistent synovitis not responsive to conventional techniques.     

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.     

Fate and effect of autogenous osteochondral fragments implanted in the middle carpal joint of horses.

Four autogenous osteochondral fragments removed from the lateral trochlear ridge of the talus were arthroscopically placed as loose bodies in a randomly selected middle carpal joint in each of 10 horses. The contralateral middle carpal joint, subjected to a sham procedure, served as control. Postoperative treatment was consistent with that for clinical arthroscopic patients. Lameness evaluation, radiographic examination, carpal circumference measurement, and synovial fluid analysis were performed before and at scheduled intervals after surgery. After a 2-month confinement, horses were subjected to an increasing level of exercise. Horses were euthanatized at intervals through 6 months. Gross and microscopic evaluations were performed on remaining fragments, articular cartilage, and synovial membrane of each middle carpal joint. Increased joint circumference, effusion, lameness, and degenerative joint disease distinguished implanted from control joints over the 6-month period. Implanted joints were characterized by grooved, excoriated cartilage surfaces, and synovium that was thick, erythematous, and irregular. At 4 weeks, implants were found to have adhered to synovium at their subchondral bone surface. The bone within fragments was undergoing necrosis, while cartilage was preserved. At 8 weeks, fragments were radiographically inapparent, grossly evident as pale plaques on the synovial surface, and composed of dense fibrous connective tissue. Synovial membrane specimens from implanted joints had inflammatory change characterized by mononuclear cell infiltration 2 months after implantation. Physical damage was apparent within articular cartilage of implanted joints at 2 months, and was significant (P less than 0.05) at 6 months after surgery. Chondrocyte degenerative change was significant (P less than 0.05) at 6 months after surgery. Focal reduction in safranin-O uptake was observed in cartilage layers adjacent to physical defects. Osteochondral loose bodies of the size implanted in the middle carpal joint of horses in this study were resorbed by the synovium within 2 months. Synovitis and significant articular cartilage damage were associated with the implanted fragments. Regardless of origin, free osteochondral fragments within the middle carpal joint should be removed, and methods to prevent residual postoperative debris should be implemented to reduce potential for articular pathologic change.     

The synovial response to exogenous phospholipid (synovial surfactant) injected into the equine radiocarpal joint compared with that to prilocaine, hyaluronan and propylene glycol

AIMS: To determine the effects of the intra-articular injection of surface-active phospholipid in a propylene glycol carrier on synovial fluid composition and joint function of horses, and to compare these effects with those observed after the intra-articular administration of prilocaine, hyaluronan and propylene glycol alone. METHODS: Twenty-four horses were randomly allocated to four treatment groups: Group 1 100 mg of surface-active phospholipid in 1 ml of propylene glycol; Group 2 1 ml of propylene glycol; Group 3 10 ml of prilocaine; Group 4 2 ml of hyaluronan. Left radiocarpal joints were injected with the treatments and the right radiocarpal joints were injected with volume-matched saline as controls. Examinations for lameness, arthrocenteses and synovial fluid analyses were performed before and at 1, 3 and 7 days after injection. RESULTS: No horses became lame but treated joints temporarily developed mild to moderate effusions. Synovial fluid analyses indicated significantly greater inflammation in treated compared to control joints and this difference was greatest 24 hours after injection. There were no differences between the four treatments based on synovial fluid analysis except for neutrophil counts and alkaline phosphatase activities, which were significantly higher in prilocaine-treated joints. CONCLUSION: In horses, the intra-articular injection of surface-active phospholipid in a propylene glycol carrier induces clinically insignificant, temporary abnormalities in synovial fluid. Surface-active phospholipid was no more injurious to the synovium than prilocaine or hyaluronan. None of the agents used in this experiment caused lameness when injected into the joints of horses. RELEVANCE: This dose and formulation appear suitable for use in future experiments investigating the efficacy of surface-active phospholipid in the treatment or prevention of osteoarthritis in horses.     

Comparison of Synovial Fluid in Middle Carpal Joints Undergoing Needle Aspiration, Infusion with Saline, and Infusion with a Combination of N-Acetyl-d-Glucosamine, Hyaluronic Acid, and Sodium Chondroitin Sulfate

Synovial fluid white blood cell (WBC) count and total protein (TP) concentration were evaluated in the midcarpal joints of horses to not only determine the effects of needle aspiration, infusion with saline, and infusion with a combination of N-acetyl-d-glucosamine, hyaluronan, and sodium chondroitin sulfate (GHCS) at two different doses to evaluate the latter for safety, but to also provide information on saline injection as a control in joints. The midcarpal joints from 24 horses were used for this study. One midcarpal joint served as an untreated control, in which only synovial fluid was aspirated, whereas the opposite joint received either 2.5 mL isotonic saline (n = 8 horses), 2.5 mL of GHCS (n = 8 horses), or 7.5 mL of GHCS (n = 8 horses). Synovial fluid WBC and TP concentration were measured on days 1, 3, 5, 7, 14, and 21. Needle aspiration caused a transient increase in synovial fluid WBC and TP levels after 1 day. Instillation of fluid (2.5 mL), whether saline or GHCS, caused significantly higher WBC and TP concentrations. GHCS at a dose of 7.5 mL created an elevation in TP level for an additional 48 hours; however, after 48 hours, WBC and TP were at concentrations that were not statistically different from controls. Even though an increase in WBC and TP concentrations occurred because of intra-articular saline and GHCS administration, these results were transient demonstrating that GHCS is no different than saline on synovial fluid, WBC, and TP parameters and that as previously described short-term elevation in synovial fluid inflammatory parameters should be expected when saline is used as a control.     

Effects of Exercise and Polysulfated Glycosaminoglycan on the Development of Osteoarthritis in Equine Carpal Joints With Osteochondral Defects

This study assessed the effects of postoperative exercise and intra-articular polysulfated glycosaminoglycan (PSGAG) on the repair of osteochondral defects in the carpal joints of ponies. Eighteen ponies with normal carpi had osteochondral defects (mean dimensions 2.4 cm × 0.9 cm) created arthroscopically on the dorsal aspect of the distal articular surface of the radial carpal bone. The ponies were randomized (while balancing for age [range, 2 to 15 years; median, 5.0 years]) to two groups—nine ponies were exercised and nine were stall confined. Beginning at surgery, six ponies in each group received five weekly intra-articular injections of PSGAG (250 mg) in one joint and lactated Ringer's solution in the contralateral joint; the remaining three ponies in each group received lactated Ringer's solution in both joints. The incremental exercise schedule on a circular, rotating walker was begun six days after surgery and occurred twice daily, reaching a maximum of 0.7 miles of walking and 2.7 miles of trotting by the third postoperative month. The effects of treatment on the joint tissues were determined by weekly lameness examinations and measurement of the range of carpal joint motion, carpal radiographs at six and 17 weeks after surgery, synovial fluid analysis, and cytologic evaluation of alcohol-fixed synovial fluid specimens at weeks 1 through 4 and week 17, and histology of the synovial membrane. Ultrasound images of the carpi were acquired before operation and at weeks 1, 2, 4, 8, 10, 13, and 17. Ponies were euthanatized 17 weeks after surgery. Exercise, without medication, caused more lameness throughout the study compared with no exercise. Exercised, nonmedicated ponies had the greatest limitation to carpal flexion (more painful joints), and nonexercised, nonmedicated (control) ponies had the least limitation to flexion. Radiographic scores indicated that the exercised, nonmedicated ponies had significantly (p < .05) more signs of osteoarthritis than exercised, medicated and control ponies. Ultrasonographic measurements indicated that exercise, without medication, caused the greatest increase in combined measurement of the joint capsule thickness and synovial fluid accumulation at all postoperative times. Synovial lining cell numbers in the synovial fluid from exercised ponies were significantly (p < .05) higher than in nonexercised ponies at week 1, and this trend continued at weeks 4 and 17 (p < .1). There were significantly (p < .05) more morphologic abnormalities in the synovial lining cells from exercised than from nonexercised ponies at week 17. Medication with PSGAG enabled exercised carpal joints to be flexed significantly further from weeks 2 through 6 compared with nonmedicated joints. Medication significantly (p < .05) reduced the combined joint capsule and synovial fluid thickness at weeks 4, 8, and 13 compared with nonmedicated joints. On histologic examination, the synovial membrane matrix of exercised, medicated joints had significantly less chronic inflammatory changes than joints receiving other treatments. The authors concluded that this level of exercise was too intense when superimposed on large osteochondral defects in the carpus because it induced osteoarthritis. Polysutfated gtycosaminoglycan ameliorated the clinical signs of osteoarthritis in the exercised ponies. However, PSGAG was also associated with the formation of cartilage repair tissue that contained less type II relative to type I collagen compared with repair tissue from nonmedicated joints.     

Pulsed radio frequency therapy of experimentally induced arthritis in ponies.

The effect of pulsed radio frequency therapy (PRFT) was evaluated on seven ponies with no arthritis and in 28 ponies in which arthritis was created using intra-articular amphotericin B to induce synovitis in the right middle carpal joint. The ponies were divided into five treatment and two control groups. Two levels of arthritis were created and two dosage levels of PRFT were evaluated. The effect of PRFT on arthritic and nonarthritic joints was measured by comparing synovial fluid parameters, the degree and duration of lameness, the range of carpal motion, and carpus circumference, for treated and untreated groups. Lesions seen radiographically, at gross pathology, and by histopathology were also compared between the treated and control groups. In the ponies with a mild form of induced arthritis, PRFT significantly (p less than 0.05) reduced the severity and duration of lameness, swelling of the carpus, and the severity of gross pathological and radiographic changes. In these ponies the synovial acid phosphatase levels were lower, the mucin clot quality was superior, and the synovial protein levels were lower for the ponies receiving PRFT as compared to the arthritic ponies receiving no treatment. A dose response effect was evident. In ponies with a slightly more severe form of arthritis, PRFT was evaluated at one dosage level. The treated ponies were significantly improved over the untreated ponies with respect to carpal range of motion, degree of lameness, carpus swelling, and radiographic lesions. No deleterious effects were noted when normal, PRFT treated, middle carpal joints were compared to contralateral untreated, normal joints. It was concluded that significant beneficial effects resulted when affected ponies were treated with PRFT.     

In vivo kinetic study on uptake and distribution of intramuscular tritium-labeled polysulfated glycosaminoglycan in equine body fluid compartments and articular cartilage in an osteochondrial defect model

The uptake and distribution of intramuscularly (IM) administered tritium-labeled polysulfated glycosaminoglycan (³H-PSGAG) in serum, synovial fluid, and articular cartilage of eight horses was quantitated, and hyaluronic acid (HA) concentration of the middle carpal joint was evaluated in a pharmacokinetic study. A full-thickness articular cartilage defect, created on the distal articular surface of the left radial carpal bone of each horse served as an osteochondral defect model. ³H-PSGAG (500 mg) was injected IM, between 14 and 35 days after creation of the defects. Scintillation analysis of serum and synovial fluid, collected from both middle carpal joints at specific predetermined times up to 96 hours post-injection, revealed mean ³H-PSGAG concentrations peaked at 2 hours post-injection. ³H-PSGAG was detected in cartilage and subchondral bone 96 hours post-injection in samples from all eight horses. There were no statistically significant differences in ³H-PSGAG concentration of synovial fluid or cartilage between cartilage defect and control (right middle carpal) joints.

HA assay of synovial fluid revealed concentrations significantly increased at 24, 48, and 96 hours post-injection in both joints. The concentration nearly doubled 48 hours post-injection. However, no statistically significant differences were found between synovial concentrations of HA in cartilage defect and control joints.

³H-PSGAG administered IM to horses, was distributed in the blood, synovial fluid, and articular cartilage. HA concentrations in synovial fluid increased after IM administration of polysulfated glycosaminoglycan.     

Effect of intra-articular gentamicin sulfate on normal equine synovial membrane

Gentamicin sulfate (3 ml; 50 mg/ml) was administered intra-articularly into 30 normal equine radiocarpal joints after arthrocentesis. Arthrocentesis alone was performed on 10 normal radiocarpal joints. Synovial fluid evaluations and gross and microscopic examinations were performed on synovial fluid and synovial membrane of designated joints at selected daily intervals over a period of 10 days. Synovial fluid from gentamicin-injected joints had greater turbidity, higher RBC and WBC counts, and higher refractive indices than did joints not injected with gentamicin. The largest increases developed on days 1 or 2 after gentamicin injection, with mean total WBC, large mononuclear cell, small mononuclear cell, and polymorphonuclear cell counts of 23,860, 11,853, 857, and 11,150 cells/microliter, respectively. Arthrocentesis alone resulted in smaller increases in these counts. Microscopic changes seen in the synovial membrane of gentamicin-injected joints included edema, leukocytic infiltration, and loss of synovial lining cells. These inflammatory changes resolved within 7 days after gentamicin injection.     

Interleukin-6 and tumour necrosis factor in synovial fluid from horses with carpal joint pathology

The carpal joints are common sites of traumatic arthritis and osteoarthritis (OA) in athletic horses. The pro-inflammatory cytokines interleukin (IL)-6 and tumour necrosis factor (TNF) may be of great importance in the development of intra-articular lesions. The aim of the present study was to investigate possible associations between synovial fluid levels of bioactive IL-6 and TNF and different types of joint lesions seen in traumatic arthritis and OA. Synovial fluid was collected from horses with carpal lameness immediately before arthroscopic surgery. Articular cartilage, synovial membranes and intra-articular ligaments were assessed macroscopically at arthroscopy. Synovial fluid levels of IL-6 and TNF were determined by bioassays, and the cytokine levels between different grades of morphologic changes in each type of assessed tissue were compared. The highest levels of IL-6 were detected in joints with chip fractures. All joints with chip fractures also showed some degree of synovitis. Tumour necrosis factor bioactivity was low and not associated with any joint lesion. Hence, TNF is not useful as a biomarker indicating a specific joint lesion in equine traumatic arthritis or OA. We conclude that a dramatic increase of IL-6 in synovial fluid indicates the presence of osteochondral fragmentation, although low or undetectable levels of IL-6 do not exclude chip fractures. The role of IL-6 in the disease process of osteochondral fragmentation needs further investigation.     

Determination of synovial fluid and serum concentrations, and morphologic effects of intraarticular ceftiofur sodium in horses

OBJECTIVES: To determine the serum and synovial fluid concentrations of ceftiofur sodium after intraarticular (IA) and intravenous (IV) administration and to evaluate the morphologic changes after intraarticular ceftiofur sodium administration. STUDY DESIGN: Strip plot design for the ceftiofur sodium serum and synovial fluid concentrations and a split plot design for the cytologic and histopathologic evaluation. ANIMALS: Six healthy adult horses without lameness. METHODS: Stage 1: Ceftiofur sodium (2.2 mg/kg) was administered IV. Stage 2: 150 mg (3 mL) of ceftiofur sodium (pHavg 6.57) was administered IA into 1 antebrachiocarpal joint. The ceftiofur sodium was reconstituted with sterile sodium chloride solution (pH 6.35). The contralateral joint was injected with 3 mL of 0.9% sterile sodium chloride solution (pH 6.35). Serum and synovial fluid samples were obtained from each horse during each stage. For a given stage, each type of sample (serum or synovial fluid) was collected once before injection and 12 times after injection over a 24-hour period. All horses were killed at 24 hours, and microscopic evaluation of the cartilage and synovium was performed. Serum and synovial fluid concentrations of ceftiofur sodium were measured by using a microbiologic assay, and pharmacokinetic variables were calculated. Synovial fluid was collected from the active joints treated during stage 2 at preinjection and postinjection hours (PIH) 0 (taken immediately after injection of either the ceftiofur sodium or sodium chloride), 12, and 24, and evaluated for differential cellular counts, pH, total protein concentration, and mucin precipitate quality. RESULTS: Concentrations of ceftiofur in synovial fluid after IA administration were significantly higher (P = .0001) than synovial fluid concentrations obtained after IV administration. Mean peak synovial fluid concentrations of ceftiofur after IA and IV administration were 5825.08 microg/mL at PIH .25 and 7.31 microg/mL at PIH 4, respectively. Mean synovial fluid ceftiofur concentrations at PIH 24 after IA and IV administration were 4.94 microg/mL and .12 microg/mL, respectively. Cytologic characteristics of synovial fluid after IA administration did not differ from cytologic characteristics after IA saline solution administration. White blood cell counts after IA ceftiofur administration were < or =3,400 cells/ML. The mean synovial pH of ceftiofur treated and control joints was 7.32 (range, 7.08-7.5) and 7.37 (range, 7.31-7.42), respectively. Grossly, there were minimal changes in synovium or cartilage, and no microscopic differences were detected (P = .5147) between ceftiofur-treated joints and saline-treated joints. The synovial half-life of ceftiofur sodium after IA administration joint was 5.1 hours. CONCLUSIONS: Synovial concentrations after intraarticular administration of 150 mg of ceftiofur sodium remained elevated above minimal inhibitory concentration (MIC90) over 24 hours. After 2.2 mg/kg IV, the synovial fluid ceftiofur concentration remained above MIC no longer than 8 hours. CLINICAL RELEVANCE: Ceftiofur sodium may be an acceptable broad spectrum antimicrobial to administer IA in septic arthritic equine joints.     

Analgesic and anti-hyperalgesic effects of epidural morphine in an equine LPS-induced acute synovitis model

Epidural morphine is widely used in veterinary medicine, but there is no information about the anti-hyperalgesic and anti-inflammatory effects in acute inflammatory joint disease in horses. The analgesic, anti-hyperalgesic and anti-inflammatory effects of epidural morphine (100mg/animal or 0.17±0.02mg/kg) were therefore investigated in horses with acute synovitis. In a cross-over study, synovitis was induced in the talocrural joint by intra-articular lipopolysaccharide (LPS). The effect of epidural morphine was evaluated using physiological, kinematic and behavioural variables. Ranges of motion (ROM) of the metatarsophalangeal and talocrural joints were measured, clinical lameness scores and mechanical nociceptive thresholds (MNTs) were assessed and synovial fluid inflammatory markers were measured. The injection of LPS induced transient synovitis, resulting in clinical lameness, decreased ranges of motion in the talocrural and metatarsophalangeal joints, decreased limb loading at rest and increased composite pain scores. Epidural morphine resulted in a significant improvement in clinical lameness, increased ROM and improved loading of the LPS-injected limb at rest, with no effects on synovial fluid inflammatory markers. Morphine prevented a decrease in MNT and, hence, inhibited the development of hyperalgesia close to the dorsal aspect of inflamed talocrural joints. This study showed that epidural morphine provides analgesic and anti-hyperalgesic effects in horses with acute synovitis, without exerting peripheral anti-inflammatory effects.     

Evaluation of the inflammatory response to two intra-articular hyaluronic acid formulations in normal equine joints

Intra-articular (IA) hyaluronic acid (HA) is commonly used to treat equine arthritis. Inflammatory response or “joint flare” is a recognized potential side effect. However, the incidence and severity of inflammation following IA HA injection in horses is not well documented. This study compared the effects of two IA HA formulations of different molecular weight (MW) and a saline control on clinical signs and synovial fluid markers of inflammation in normal equine joints. Eight adult horses each had three healthy fetlock joints randomly assigned to treatment with either 1.4 mega Dalton HA, 0.8 mega Dalton HA or saline control once weekly for three weeks. Clinical evaluation and synovial fluid analysis were performed by blinded assessors. Outcomes of interest were lameness score, joint effusion score and synovial fluid white cell count and differential, total protein, viscosity and serum amyloid A. Joints injected with HA developed significant mild-to-moderate inflammatory responses often associated with lameness and joint effusion compared with saline control joints. The higher MW HA formulation elicited a significantly greater inflammatory response than the lower MW HA after the first injection. In HA injected joints, viscosity remained poor for the entire study. Both IA HA formulations in this study induced an inflammatory response in healthy equine joints. This may have implications for the use of HA in equine joints. The findings in this study are limited to the two HA formulations used. Further investigation of different HA formulations and the use of HA in normal and arthritic equine joints is warranted.     

Evaluation of iatrogenic hemarthrosis of the metacarpophalangeal joint as a method of induction of temporary reversible lameness in horses

OBJECTIVE: To determine whether iatrogenic hemarthrosis of the metacarpophalangeal joint could be used as a model for temporary reversible joint pain in horses. ANIMALS: 8 adult horses. PROCEDURE: Each horse was evaluated on a treadmill before and after injection of 1 metacarpophalangeal joint with 10 mL of autogenous blood. Horses were evaluated subjectively and objectively by use of a computerized force measurement system at intervals until lameness abated. The mean force difference between injected and noninjected limbs at all time periods after injection was compared with the difference between limbs at baseline. From each horse, synovial fluid samples collected before and 24 hours and 30 days after injection were analyzed for total protein concentration and cell type and number. Venous blood samples were collected before and 6 and 24 hours after injection for assessment of plasma cortisol concentration. RESULTS: For 24 hours after injection, the mean force difference between injected and noninjected limbs was significantly increased over baseline. The greatest force difference was detected after 2 and 4 hours. Baseline and 24-hour force data were not significantly different. Compared with baseline values, synovial fluid protein concentration and nucleated cell and RBC counts were increased significantly at 24 hours after injection but were not different at 30 days after injection. No significant changes in plasma cortisol concentration were detected at any time point. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, iatrogenic hemarthrosis of the metacarpophalangeal joint appears to induce temporary reversible lameness with a mild to moderate degree of synovitis.     

Evaluation of safety and pharmacokinetics of vancomycin after intravenous regional limb perfusion in horses

OBJECTIVE: To evaluate clinical variables, regional concentrations, and pharmacokinetics of vancomycin in the synovial fluid of distal forelimb joints of horses after IV regional limb perfusion. ANIMALS: 6 horses. PROCEDURE: Vancomycin was administered via IV regional limb perfusion to the distal portion of the forelimbs of anesthetized horses. Drug (300 mg of vancomycin hydrochloride in 60 mL of saline [0.9% NaCl] solution) was infused into 1 forelimb, whereas the contralateral limb served as a control and was perfused with 60 mL of saline solution. Solutions were injected into the lateral digital vein after digital exsanguination. Synovial fluid from the metacarpophalangeal (MTCP) and distal interphalangeal (DIP) joints and systemic blood were collected prior to perfusion and 15, 30, 45, 65, and 90 minutes after initiation of the infusion. Synovial fluid from the MTCP joint and blood were also obtained at 4, 8, 12, and 24 hours after infusion. Plasma urea and creatinine concentrations, degree of lameness, and certain clinical variables involving the MTCP joint and infusion site were assessed for 7 days. Results were compared between the vancomycin treatment and control groups. RESULTS: No complications or significant differences in renal function, lameness, or clinical variables were observed between groups. Vancomycin concentrations exceeded 4 microg/mL in MTCP joints for approximately 20 hours. Higher concentrations were reached in DIP joints than in MTCP joints. CONCLUSIONS AND CLINICAL RELEVANCE: IV regional limb perfusion with 300 mg of vancomycin as a 0.5% solution was safe and may be useful in horses as treatment for distal limb infections.