Responses to an intra-articular lipopolysaccharide challenge following dietary supplementation of Saccharomyces cerevisiae fermentation product in young horses

Dietary intervention may be a valuable strategy to optimize the intra-articular environment in young horses to prolong their performance career. To test the hypothesis that dietary supplementation of a Saccharomyces cerevisiae fermentation product would reduce markers of joint inflammation and increase markers of cartilage metabolism following a single inflammatory insult, Quarter Horse yearlings (mean ± SD; 9 ± 1.0 mo) were balanced by age, sex, body weight (BW), and farm of origin and randomly assigned to the following treatment groups: 1.25% BW/d (dry matter basis) custom-formulated concentrate only (CON; n = 9) or concentrate top-dressed with 21 g/d S. cerevisiae fermentation product (SCFP; n = 10) for 98 d. Horses had ad libitum access to Coastal bermudagrass hay. On day 84, one randomly selected radial carpal joint from each horse was injected with 0.5 ng lipopolysaccharide (LPS) solution. The remaining carpal joint was injected with sterile lactated Ringer’s solution as a contralateral control. Synovial fluid obtained before supplementation (day 0) and on day 84 at preinjection hour 0 and 6, 12, 24, 168, and 336 h postinjection was analyzed for prostaglandin E₂ (PGE₂), carboxypropeptide of type II collagen (CPII), and collagenase cleavage neopeptide (C2C) by commercial assays. Rectal temperature, heart rate, respiration rate, carpal surface temperature, and carpal circumference were recorded prior to each sample collection and for 24 h postinjection. Data were analyzed using linear models with repeated measures. From day 0 to 84, synovial C2C declined (P ≤ 0.01) and the CPII:C2C ratio increased (P ≤ 0.01) in all horses with no effect of diet. In response to intra-articular LPS, synovial PGE₂ increased by hour 6 (P ≤ 0.01) and returned to baseline by hour 336; CPII increased by hour 12, remained elevated through hour 168 (P ≤ 0.01), and returned to baseline by hour 336; and C2C increased by hour 6 (P ≤ 0.01) but did not return to baseline through hour 336 (P ≤ 0.01). Post-intra-articular injection, PGE₂ levels were lower in SCFP than CON horses (P = 0.01) regardless of injection type. Synovial CPII and the CPII:C2C ratio demonstrated stability during the LPS challenge in SCFP compared with CON horses (P ≤ 0.01). Clinical parameters were not influenced by diet but increased in response to repeated arthrocentesis (P ≤ 0.01). Dietary SCFP may favorably modulate intra-articular inflammation following an acute stressor and influence cartilage turnover in young horses.     

Effects of a Novel Dietary Supplement on Indices of Muscle Injury and Articular GAG Release in Horses

This study determined the ability of an oral nutraceutical supplement to attenuate the oxidative stress and inflammation that occurs in muscles and joints with repeated bouts of high-intensity exercise in horses. The supplement, fed daily, was comprised of whole dried mushrooms, golden flaxseed, omega-3 fatty acids, plant-based enzymes, a melon-concentrate powder, and Saccharomyces cerevisiae boulardii. Ten horses participated in a partial cross-over design, with 7 horses completing the Control trial and 7 horses completing the supplement trial. Blood and synovial fluid samples (from the intercarpal joint) were taken before, and at 1 and 24 hours after a standardized, repeated high-intensity exercise test that was performed before supplementation and on the 22nd day of supplementation. At the end of the supplement trial exercise resulted in reduced concentrations of plasma markers of oxidative stress (decreased thiobarbituric acid reactive substances, with increased total antioxidant status and increased superoxide dismutase activity); there was no effect on plasma markers of muscle injury (creatine, creatine kinase, and aspartate aminotransferase) or inflammation (PGE₂, nitric oxide). Within synovial fluid, there was a tendency for increased superoxide dismutase activity, and decreased concentration of glycosaminoglycans. It is concluded that the supplement, when fed to horses as part of the normal diet for 23 days, was associated with reduced concentrations of markers of oxidative stress and inflammation in muscle and synovial fluid.     

Cartilage oligomeric matrix protein and hyaluronan levels in synovial fluid from horses with osteoarthritis of the tarsometatarsal joint compared to a control population

REASONS FOR PERFORMING STUDY: Quantification of cartilage oligomeric matrix protein (COMP) levels within synovial fluid from the tarsometatarsal joint has not previously been reported and an effective synovial fluid marker would allow monitoring of disease progression and treatment. OBJECTIVES: To quantify levels of COMP and hyaluronan (HA) in synovial fluid from the tarsometatarsal joint, identify differences in levels from horses with osteoarthritis (OA) of the tarsometatarsal joint compared to a control population and to correlate levels with radiographic changes in horses with OA. METHODS: Synovial fluid was collected from the tarsometatarsal joint of 25 horses without hindlimb lameness (controls) and 25 lame horses, subjected to analgesia of the joint. COMP concentrations were measured using a homologous inhibition ELISA. Immunoblots of synovial fluid from 3 lame horses and 3 controls were performed to identify fragmentation of COMP. Hyaluronan (HA) concentration in synovial fluid was determined using a competition ELISA. Radiographs of the lame horses with OA were scored and correlated with levels of COMP and HA. RESULTS: Concentrations of COMP in OA of the tarsometatarsal joint were significantly lower than in the control samples. An additional fragment band of COMP (approximately 30 kDa) was identified on the immunoblots of the horses with OA and this fragment was not identified in controls. No significant difference was identified in the HA or HA:COMP ratio between lame and control horses. There was no correlation between levels of synovial fluid COMP and HA, and radiographic changes. CONCLUSIONS AND POTENTIAL RELEVANCE: Lowered levels of COMP in synovial fluid of tarsometatarsal joints correlates with the presence of osteoarthritis. However, a single value cannot be used to stage the disease process. Levels of HA may not be a useful marker for this disease. Decreased, rather than increased COMP levels, may reflect significant loss of cartilage in established osteoarthritis. A specific assay for the COMP fragment generated with osteoarthritis may allow the earlier detection of clinical cases.     

A Marine Mineral Supplement Alters Markers of Bone Metabolism in Yearling Arabians

This study tested whether the supplement (Aquacid), high in calcium and other minerals, can alter markers of bone metabolism and mineralization of the equine third metacarpus bone. Radiographs were taken of the left third metacarpus of 14 yearlings. Radiographic bone aluminum equivalence (RBAE) of each cortex was calculated to estimate mineral content. Blood samples were also taken at this time. Horses were ranked according to RBAE and gender, were pair-matched, and randomly assigned to two treatment groups. Each group was provided one of two mineral supplements in addition to their normal diet. The treated group (Aq) received 75 g Aquacid/horse/d, which provided an additional 15 g of calcium. The control group (Co) received 39.5 g of limestone to provide similar amounts of calcium. The study lasted for 112 days, with blood being taken every 28 days. At day 56 and 112, additional radiographs was taken to track changes in RBAE. Blood was analyzed for osteocalcin (a bone formation marker) and serum C-telopeptide crosslaps of type I collagen (a bone resorption marker) to detect alterations in bone metabolism. Using day 0 values as a covariate for bone markers, there was a trend (P = .07) for osteocalcin concentrations to be greater in Aq horses than in Co. Likewise, C-telopeptide crosslaps of type I collagen concentrations were greater (P < .0001) in Aq horses than in Co. There were minimal differences in RBAE values. These findings suggest Aquacid, while not altering bone mass, increases bone turnover and may aid in repairing damaged bone and preventing injuries.     

Increased plasma silicon concentrations and altered bone resorption in response to sodium zeolite a supplementation in yearling horses

This study examined the effect of supplementation of a bioavailable source of silicon (sodium zeolite A) on altering systemic markers of bone metabolism in horses. Twenty yearlings (ten Quarter Horses and ten Arabians) were randomly grouped as silicon (Si) supplemented (S; n=10), in which yearlings consumed 2% of the total diet as a Si-containing supplement, and a second non-supplemented control group (C; n=10). Blood samples were taken on days 0, 15, 30 and 45. Both plasma and serum were collected; the plasma was analyzed for Si concentrations and serum was analyzed for osteocalcin (OC), carboxy-terminal pyridinoline cross-linked telopeptide region of type I collagen (ICTP), and pyridinoline and deoxypyridinoline crosslinks (PYD). Supplemented yearlings had higher plasma Si concentrations than C yearlings by day 15, and remained higher than C yearlings on days 30 and 45 (P < 0.0001 for all days). There were no differences between treatment groups for OC or PYD concentrations (P > .05); however, ICTP concentrations were lower in S yearlings on day 45 when compared to C yearlings (P = .04). Results indicate that sodium zeolite A supplementation (consumed at 2% of the total diet) increases plasma Si concentrations. Furthermore, results indicate that Si-supplemented yearlings may have decreased bone resorption, which may provide for greater net bone formations, as OC concentrations were not different between groups. Unfortunately, systemic markers give no indication as to the quality of the bone that may be formed, and further research in the area of Si supplementation, bone metabolism and bone strength is required to establish conclusive evidence as to the benefits of supplemental Si to the skeletal system.     

Mineral balance in horses fed two supplemental silicon sources

Numerous studies suggest that silicon (Si) supplementation is beneficial for mineral metabolism and bone health. Mineral balance studies have not been performed in horses to determine how these supplements affect absorption of other minerals. The purpose of these studies was to investigate the effects of two different Si supplements on mineral absorption and retention in horses. Eight geldings were randomly placed in one of two groups: control (CO) or supplemental Si, which was provided by one of two supplements. The first, sodium aluminium silicate (SA), contains a bioavailable form of Si and is high in aluminium (Al). The second supplement contains oligomeric orthosilicic acid (OSA). All horses received textured feed and ad libitum access to hay. Supplemented horses received either 200 g of SA or 28.6 ml of OSA per day. Following a 10-day adaptation period, the horses underwent a 3-day total collection. Blood samples were taken on days 0 and 13. The two balance studies were conducted 4 months apart to reduce carryover effects. Intakes of Al and Si were greater with SA supplementation (p < 0.05). Sodium aluminium silicate increased faecal and urinary Si excretion (p < 0.05). Calcium retention and apparent digestion were increased by SA (p < 0.05). It also maintained plasma Si compared with the CO which tended to have a decrease in plasma Si (p = 0.08). Supplemental OSA increased retention of Ca and B (p < 0.05) and apparent digestion of B (p < 0.01). Orthosilicic acid tended to increase Si retention (p = 0.054), apparent digestion (p < 0.065), and also increased plasma Si. Both supplements were able to alter Ca retention and B metabolism, however, only OSA was able to alter Si retention, digestibility and plasma concentration. Orthosilicic acid, an Si supplement without substantial Al, appears to be a viable option for Si supplementation as it increased Si retention and digestibility.     

Evaluation of avocado and soybean unsaponifiable extracts for treatment of horses with experimentally induced osteoarthritis

OBJECTIVE: To evaluate the use of a combination of avocado and soybean unsaponifiable (ASU) extracts for the treatment of experimentally induced osteoarthritis in horses. ANIMALS: 16 horses. PROCEDURES: Osteoarthritis was induced via osteochondral fragmentation in 1 middle carpal joint of each horse; the other joint underwent a sham operation. Horses were randomly allocated to receive oral treatment with ASU extracts (1:2 [avocado-to-soybean] ratio mixed in 6 mL of molasses; n = 8) or molasses (6 mL) alone (placebo treatment; 8) once daily from days 0 to 70. Lameness, response to joint flexion, synovial effusion, gross and histologic joint assessments, and serum and synovial fluid biochemical data were compared between treatment groups to identify effects of treatment. RESULTS: Osteochondral fragmentation induced significant increases in various variables indicative of joint pain and disease. Treatment with ASU extracts did not have an effect on signs of pain or lameness; however, there was a significant reduction in severity of articular cartilage erosion and synovial hemorrhage (assessed grossly) and significant increase in articular cartilage glycosaminoglycan synthesis, compared with placebo-treated horses. CONCLUSIONS AND CLINICAL RELEVANCE: Although treatment with ASU extracts did not decrease clinical signs of pain in horses with experimentally induced osteoarthritis, there did appear to be a disease-modifying effect of treatment, compared with findings in placebo-treated horses. These objective data support the use of ASU extracts as a disease-modifying treatment for management of osteoarthritis in horses.     

Clinical, biochemical, and histologic effects of intra-articular administration of autologous conditioned serum in horses with experimentally induced osteoarthritis

OBJECTIVE: To assess the clinical, biochemical, and histologic effects of intra-articular administration of autologous conditioned serum (ACS) in the treatment of experimentally induced osteoarthritis in horses. ANIMALS: 16 horses. PROCEDURES: Osteoarthritis was induced arthroscopically in 1 middle carpal joint of all horses. In 8 placebo- and 8 ACS-treated horses, 6 mL of PBS solution or 6 mL of ACS was injected into the osteoarthritis-affected joint on days 14, 21, 28, and 35, respectively; PBS solution was administered in the other sham-operated joints. Evaluations included clinical assessment of lameness and synovial fluid analysis (performed biweekly); gross pathologic and histologic examinations of cartilage and synovial membrane samples were performed at necropsy. RESULTS: No adverse treatment-related events were detected. Horses that were treated with ACS had significant clinical improvement in lameness, unlike the placebo-treated horses. Among the osteoarthritis-affected joints, ACS treatment significantly decreased synovial membrane hyperplasia, compared with placebo-treated joints; although not significant, the ACS-treated joints also appeared to have less gross cartilage fibrillation and synovial membrane hemorrhage. The synovial fluid concentration of interleukin-1 receptor antagonist (assessed by use of mouse anti-interleukin-1 receptor antagonist antibody) was increased following treatment with ACS. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this controlled study indicated that there was significant clinical and histologic improvement in osteoarthritis-affected joints of horses following treatment with ACS, compared with placebo treatment. On the basis of these findings, further controlled clinical trials to assess this treatment are warranted, and investigation of the mechanisms of action of ACS should be pursued concurrently.     

Metalloproteinases and tumor necrosis factor-alpha activities in synovial fluids of horses: correlation with articular cartilage alterations

Early detection of osteoarthritis in horses represents a challenge for equine practitioners. Several biological markers have been implicated in the pathological processes involved in articular cartilage destruction. To further document cartilage matrix proteases production, synovial fluid was collected from 14 horses (90 joints) before they were subjected to euthanasia. Growth macroscopic examination of the joints gave information on cartilage alterations. Samples were analyzed for matrix metalloproteinase (MMPs) activities by gelatin zymography and tumor necrosis factor alpha (TNF-alpha) cytotoxicity using L929 cells. Significant increase of MMP-9 monomer and dimer were found in synovial fluids of joints with severe cartilage alterations. On the contrary, the activity of TNF-alpha was not correlated to the degree of joint damage. The levels of MMP-9 monomer and dimer in the synovial fluid could reflect cartilage alteration in arthritis in the horse.     

Efficacy of an Extract of Blue-Green Algae in Amelioration of Lameness Caused by Degenerative Joint Disease in the Horse

Degenerative joint disease (DJD) or osteoarthritis in the equine is largely a result of biomechanical stressors that result in inflammation within the joint, which with continual exposure, leads to progressive degeneration. A myriad of therapies are available for treatment of horses with DJD including nonsteroidal anti-inflammatory drugs, corticosteroids, physiological modifiers, and biological therapies. C-phycocyanin, a protein-bound pigment found in blue-green algae, has been shown to have anti-inflammatory and antioxidant properties and is available as a diet supplement in people, dogs, and horses. Forty-one horses with naturally occurring lameness as a result of DJD were selected for this study. In a blinded study, horses were randomly assigned to either the commercial or placebo product that was fed once daily for 12 months. At selected time points, lameness evaluation was performed through objective gait analysis. Previous medical records for management of lameness were reviewed for all enrolled horses. No statistically significant differences existed within or between the two groups when gait was evaluated objectively nor within individual horses between time points. Review of medical records found that five treated horses had a decreased in frequency of intra-articular injection of corticosteroids. Although this study did not find statistically significant improvement of lameness with oral daily supplementation of C-phycocyanin in equine athletes, there appeared to be a trend of improvement in lameness during the loading period and a decrease in frequency of administration of intra-articular corticosteroids. Further investigations using a higher dose seem warranted.     

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.     

Osteochondral Fragmentation in the Synovial Pad of the Fetlock in Warmblood Horses

Objectives— To determine clinical and arthroscopic characteristics associated with fragments in the synovial pad of the fetlock and to characterize their morphology. Study Design— Retrospective study. Animals— Warmblood horses (n=104) with fragment(s) in the synovial pad. Methods— S ignalment and results of radiographic and clinical examination were collected before surgery. After arthroscopic fragment removal and joint evaluation for synovial and/or cartilage abnormalities, fragments were measured, and evaluated by histopathology. Results— Synovial pad fragments (n=142) were removed from 127 fetlocks. Two older horses had lameness. During arthroscopy, abnormalities were observed in 40 joints (31.5%) and multivariate logistic regression analysis showed a significant correlation between the observed arthroscopic abnormalities and the presence of large fragments (P=.016). Fragments were osteochondral bodies completely surrounded by fibrous tissue. At the edges of the hyaline cartilage cap an underlying fibrous structure was obvious in the extracellular matrix giving it a reactive pattern. Conclusions— Although the impact on lameness was minimal, there was a significant correlation between arthroscopic abnormalities and presence of large synovial pad fragments. On histopathology, these osteochondral fragments are embedded in fibrous tissue and show a reactive pattern. They are not a manifestation of any well‐described joint pathology. Clinical Relevance— Large synovial pad fragments in Warmblood horses can be associated with synovial and cartilage abnormalities, but further studies are warranted to determine their origin and clinical importance.     

Recombinant equine growth hormone administration: effects on synovial fluid biomarkers and cartilage metabolism in horses

REASONS FOR PERFORMING STUDY: Recombinant equine growth hormone (reGH) has recently been evaluated for effects on body condition and wound healing. It has the potential to influence articular cartilage via stimulation of IGF-1. OBJECTIVES: To investigate effects of administration on synovial joint metabolism. METHODS: Six mature horses were given 20 microg/kg bwt reGH daily for 8 weeks by i.m. injection. Three control horses were injected with sterile water. Serum and synovial fluid samples were collected at 6, 8, 11 and 16 weeks for GH and IGF-1 assays. Articular cartilage harvested at week 16 was evaluated by Western analysis using monoclonal antibodies BC-13, BC-4, 8-A-4 and CH-3. RESULTS: Concentrations of IGF-1 in serum and synovial fluid were significantly elevated (P < 0.05) at 6 and 8 weeks in the reGH group. Glycosaminoglycan concentrations in synovial fluid were significantly less than controls at these time points, suggesting that reGH may modulate proteoglycan metabolism in articular cartilage. In the reGH group, there were not any alterations in synovial fluid content of 3B3(-) epitope or aggrecan metabolite, or in aggrecan or link protein catabolites retained within cartilage, that might be expected with development of osteoarthritis. CONCLUSIONS: Intramuscular administration of reGH may be a more efficient means of delivery of IGF-1 to joints for cartilage resurfacing initiatives. POTENTIAL RELEVANCE: We found no alterations in cartilage metabolism indicative of development of osteoarthritis.     

Effects of intra-articular administration of dimethylsulfoxide on chemically induced synovitis in immature horses

The effects of intra-articular administration of dimethylsulfoxide (DMSO) on chemically induced synovitis in the middle carpal joint of 6 weanling horses were evaluated. Following aseptic collection of synovial fluid, the middle carpal joint of each forelimb was injected with 50 mg of Na-monoiodoacetate to induce synovitis. Eight days after injection, synovial fluid was obtained and the right middle carpal joints were injected with 2 ml of 40% DMSO in lactated Ringer solution. The corresponding joints of the left limb (control) were injected with 2 ml of lactated Ringer solution. Sampling and treatments were repeated on post-injection days 11 and 14, for a total of 3 treatments. Horses were visually evaluated daily for lameness and joint effusion. Synovial fluid was evaluated for color and clarity, differential and total WBC count, total protein content, and hyaluronic acid concentration. The Kaegi gait analysis system provided an objective assessment of lameness prior to inducing synovitis, again on day 7, and on day 17. At necropsy (day 17), synovial fluid, synovial membrane, and articular cartilage specimens were collected. Joint effusion was evident 12 hours after injection of Na-monoiodoacetate in all joints. Mild lameness was evident at 24 hours; however, the lameness resolved by 72 hours. Objective assessment of lameness did not reveal significant differences between treatment or control limbs. Hyaluronic acid concentrations increased significantly (P = 0.023) above baseline values in most joints over the study period. Synovial fluid WBC counts increased significantly (P = 0.002) following Na-monoiodoacetate injection and remained significantly (P = 0.002) above baseline values throughout the study.(ABSTRACT TRUNCATED AT 250 WORDS)     

Long-Term Outcome in 44 Horses with Stifle Lameness After Arthroscopic Exploration and Debridement

Objective— To (1) examine the outcome in horses with osteoarthritis or intra‐articular soft tissue injuries of the stifle after arthroscopic exploration and debridement and (2) to determine any imaging or surgical findings that may influence prognosis. Design— Case series. Animals— Horses (n=44) with lameness referable to the stifle, diagnosed with osteoarthritis, meniscal tears, or other intra‐articular soft tissue injuries based on arthroscopic examination. Methods— Medical records of horses with stifle lameness that had arthroscopic exploration were reviewed. Horses with osteochondrosis lesions, intra‐articular fractures, or osseous cyst‐like lesions were excluded. Pertinent case information was analyzed and short‐ and long‐term outcome was assessed. Results— There was no association between radiographic score and surgery score. Diagnostic ultrasound had a sensitivity of 79% and a specificity of 56% for identifying meniscal injuries. Follow‐up information was available for 35 horses; 23 horses (60%) improved after surgery, 16 (46%) became sound, and 13 (37%) returned to their previous level of function. A negative association was observed between age and degree of preoperative lameness and outcome. More severe changes observed on preoperative radiographs were also negatively associated with prognosis. No horses with grade 3 meniscal tears improved postoperatively and increasing meniscal pathology was negatively associated with return to previous function. A weak association between surgery grade and outcome was also observed. Degree of chondral damage, location of primary pathology, and microfracture techniques had no effect on outcome. Conclusions— Advanced horse age, severe lameness and preoperative radiographic changes, and presence of large meniscal tears are associated with a negative postoperative outcome for horses with stifle lameness. Appearance of the articular surface at surgery appears to be an inconsistent prognostic indicator. Clinical Relevance— Some horses with extensive cartilage damage may return to athletic function after arthroscopic debridement and lavage. A more pessimistic prognosis may be given to older horses, those with more severe preoperative lameness, and those with severe radiographic changes or large meniscal tears.     

Assessment of the effects of age and joint disease on hydroxyproline and glycosaminoglycan concentrations in synovial fluid from the metacarpophalangeal joint of horses

OBJECTIVE: To assess the effects of age and joint disease on hydroxyproline and glycosaminoglycan (GAG) concentrations in synovial fluid from the metacarpophalangeal joint of horses and evaluate the association of those concentrations with severity of osteoarthritis and general matrix metalloproteinase (MMP) activity. SAMPLE POPULATION: Synovial fluid was collected from the metacarpophalangeal joints of foals at birth (n = 10), 5-month-old foals (10), 11-month-old foals (5), and adult horses (73). PROCEDURE: Hydroxyproline and GAG concentrations were determined in synovial fluid samples. The severity of osteoarthritis in adult joints was quantified by use of a cartilage degeneration index (CDI) and assessment of general MMP-activity via a fluorogenic assay. RESULTS: Hydroxyproline and GAG concentrations in synovial fluid were highest in neonates and decreased with age. Concentrations reached a plateau in adults by 4 years and remained constant in healthy joints. In synovial fluid from osteoarthritic joints, hydroxyproline and GAG concentrations were not increased, compared with unaffected joints, but hydroxyproline were significantly correlated with the CDI and general MMP activity. There was no significant correlation between GAG concentration and CDI value or MMP activity. CONCLUSIONS AND CLINICAL RELEVANCE: Changes in hydroxyproline concentration in synovial fluid appeared to indicate damage to collagen of the articular cartilage. In joints with osteoarthritis, the lack of high GAG concentration in synovial fluid and the absence of a significant correlation between GAG concentration and CDI values or MMP activity may severely limit the usefulness of this marker for monitoring equine joint disease.     

Effect of Betamethasone and Exercise on Equine Carpal Joints With Osteochondral Fragments

Osteochondral fragments were created arthroscopically on the distal aspect of both radial carpal bones in 12 horses. On day 14 after surgery, one middle carpal joint of each horse was injected with 2.5 mL Betavet Soluspan (3.9 mg betamethasone sodium phosphate and 12 mg betamethasone acetate per milliliter) and the contralateral joint was injected with 2.5 mL saline as a control. Intra-articular treatments were repeated on day 35. On day 17, six horses began exercising 5 days per week on a high-speed treadmill. The other six horses were kept in box stalls throughout the study as nonexercised controls. On day 56, all horses were examined clinically and radiographically and then were euthanatized. Samples were obtained for histological, his-tochemical, and biochemical evaluation. Mild lameness was observed in five of the six exercised horses at day 56; four horses were lame in the control limb and one horse was lame in the treated limb. Of the five nonexercised horses evaluated for lameness, two were lame in the control limb, two were lame in the treated limb, and one was lame in both the control and the treated limb. No differences were noted on radiographs or palpation of steroid treated limbs versus control limbs. Firm reattachment of the osteochondral fragment to the radial carpal bone occurred in all but three joints. Gross cartilage damage was not different between steroid-treated joints and joints injected with saline. Histologically, there were no significant detrimental effects of betamethasone with or without exercise, but there was a tendency for more pathological change in treated joints. There was a trend toward decreased glycosaminoglycan staining in steroid treated joints of rested horses, whereas exercised horses had similar glycosaminoglycan staining in treated and control joints. No significant difference in the water content or uronic acid concentration was detected between treated and control joints. Intra-articular betamethasone administration in this carpal chip model was not associated with any significant detrimental effects in either rested or exercised horses.     

Scanning electron microscopic findings of the navicular bone and deep flexor tendon in podotrochlosis of horses]

In 25 adult horses the podotrochlea of 49 forelimbs was examined by means of light and electron microscopy in order to correlate clinical and morphological findings. According to the clinical diagnosis the animals were divided into three groups: lameness due to syndrome of podotrochlosis (group 1) or due to tendopathy (group 2) and horses without lameness (group 3). The most striking pathological findings of the navicular bone and the opposite surface of the deep flexor tendon were found in horses with podotrochlosis, consisting of loss of cartilage and tendon matrix with denudation of collagen fibrils, superficial degeneration and necrosis, focal occurrence of fissures and far-reaching defects, sometimes accompanied by adhesions between tendon and navicular bone surface. In radiologically only slightly altered tendons and navicular bones light and scanning electron microscopy often revealed moderate to severe pathological changes, which sometimes could also be seen in clinically healthy horses. The distribution of the lesions indicate that in course of podotrochlosis the initial alterations caused by repeated microtraumata start at the surface of the deep flexor tendon and subsequently spread to the opposite facies flexoria of the navicular bone. An involvement of the cartilage and the synovial layer of the hoof joint in cases with podotrochlosis could only be rarely observed and is to regard as a secondary manifestation.