Use of allogeneic freeze-dried conditioned serum for the prevention of degradation in cartilage exposed to IL-1ß.

BACKGROUND: Autologous conditioned serum (ACS) has been extensively used in the field of veterinary orthopaedics and sports medicine. Due to the autologous and blood-derived nature of this product, issues such as individual variability, need for storage at low temperatures and non-availability for immediate are frequently encountered for ACS use in the field. To address those issues, we proposed the evaluation of an off-the-shelf allogeneic freeze-dried version of conditioned serum in an in vitro model of osteoarthritis. In this study, we evaluated if origin (autologous and allogeneic) and preparation (frozen and freeze-dried) of conditioned serum could influence in its effect in an in vitro model. RESULTS: IL-1ß stimulation in cartilage led to a significant increase in media GAG and decreased levels of GAG in cartilage explants at the termination of the experiment. No significant differences were noted in outcomes measured in the cartilage explants with respect to the main effects of treatment (frozen versus freeze-dried serum), autologous versus allogeneic preparations or based on serum concentration. CONCLUSIONS: The study did not observe any substantial differences in the response of cartilage to allogeneic freeze-dried CS when compared to other independent parameters (autologous and frozen preparations). Further investigation using in vivo systems appears warranted.

Current use of biologic therapies for musculoskeletal disease: A survey of board-certified equine specialists.

OBJECTIVE: To evaluate the use of mesenchymal stem cells (MSCs), autologous conditioned serum (ACS), platelet-rich plasma (PRP), and autologous protein solution (APS) for the treatment of equine musculoskeletal disease by diplomates of the American College of Veterinary Surgery (ACVS), and American College of Veterinary Sports Medicine and Rehabilitation (ACVSMR). STUDY DESIGN: Cross-sectional study. SAMPLE POPULATION: Diplomates (n = 423). METHODS: An email link was sent to ACVS and ACVMR diplomates. A survey contained 59 questions regarding demographics, as well as indications, frequency, adverse effects, and limitations of use. Responses were analyzed using Fisher's exact test. RESULTS: One hundred and fifty four surveys were analyzed. Years in practice and type of practice were not associated with biologic therapy use. PRP was the most used therapy (120/137; 87.5%). PRP and MSCs were most often administered intralesionally while ACS and APS were most often administered intra-articularly. ACS (50/104; 48.1%) treatment was repeated commonly within 2 weeks of initial injection. MSCs (39/90; 43.3%) and PRP (38/100; 38%) were commonly repeated 1-2 months after initial injection and APS was typically repeated >4 months after initial injection (21/53; 39.6%). Local inflammation and expense were the most common adverse effect and limitation of use. CONCLUSION: Diplomates most commonly utilized PRP and MSC intralesionally for soft-tissue injuries, and ACS and ACP intra-articularly for joint injury. Protocols for repeated administration varied widely. Local inflammation was a clinical concern with the use of biologics. CLINICAL SIGNIFICANCE: Biologic therapies are used commonly by ACVS and ACVSMR diplomates for soft tissue and joint disease.

Physiologic effects of long-term immobilization of the equine distal limb.

OBJECTIVE: To describe the effects of distal limb immobilization and remobilization in the equine metacarpophalangeal joint. STUDY DESIGN: Randomized, prospective experimental study. ANIMALS: Eight healthy, skeletally mature horses. METHODS: One forelimb of each horse was immobilized in a fiberglass cast for 8 weeks; this was followed by 12 weeks of a treadmill-based training program after the cast had been removed. Clinical examinations, radiography, computed tomography (CT), nuclear scintigraphy, MRI, and histomorphometry were used to examine the third metacarpal (MC3), proximal phalanx, proximal sesamoid bones, and associated soft tissues in each horse. Serum and synovial fluid were collected for biomarker analyses. RESULTS: Distal limb immobilization resulted in persistent lameness (P < .001), effusion (P = .002), and a decreased range of motion (P = .012) as well as radiographically visible fragments (P = .036) in the cast forelimb. Bone density was decreased (P < .001) in MC3 according to CT, and trabecular bone fluid was increased (P < .001) according to MRI in the cast forelimb. The cast forelimbs had a change (P = .009) in the appearance of the deep digital flexor tendon according to MRI immediately after removal of the cast. Numerous clinical, radiographic, CT, and MR abnormalities were visible at the end of the study period. CONCLUSION: Eights weeks of cast immobilization induced changes in bone, cartilage, and periarticular soft tissues that were not reversed after 12 weeks of remobilization. CLINICAL SIGNIFICANCE: Cast application should be used judiciously in horses with musculoskeletal injuries, balancing appropriate stabilization with potential morbidity secondary to cast placement.

Evaluation of allogeneic freeze-dried platelet lysate in cartilage exposed to interleukin 1-ß in vitro.

BACKGROUND: Platelet-rich plasma (PRP) as well as other platelet-derived products have been used as a potential disease-modifying treatment for musculoskeletal diseases, such as osteoarthritis (OA). The restorative properties of such products rely mainly on the high concentrations of growth factors, demonstrating encouraging results experimentally and clinically. Yet, the autologous blood-derived nature of the PRP product lead to limitations that precludes it's widespread use. The main limitations for PRP use are; product variability, the need for minimum laboratory settings in most cases, and the need for storage at low temperatures to preserve its properties. Based on these limitations, the objective of this study was to investigate an allogeneic off-the-shelf platelet lysate (PL) in cartilage exposed to interleukin 1ß (IL-1ß). For this purpose, blood and cartilage were harvested from eight skeletally mature and healthy horses. Blood was processed into PL aliquots and divided into three groups (Frozen, Freeze-dried and Filtered freeze-dried), used in autologous and allogeneic conditions and in three different concentrations (1.5, 3 and 6-fold). Different PL preparations were then applied in cartilage culture with interleukin-1 beta and cultured for 10 days. Cartilage and media samples were collected and analyzed for total GAG and 35SO4-labeled GAG content. RESULTS: No significant differences between the controls and PL groups in cartilage and media were demonstrated. The effects of PL on cartilage matrix were concentration dependent and intermediate concentrations (3-fold) in PL showed increased 35SO4-labelled GAG in cartilage. CONCLUSION: In conclusion, the allogeneic freeze-dried PL presented equivalent effects compared to frozen autologous PL. Intermediate platelet concentration on average demonstrated improved results, demonstrating less GAG loss compared to other concentrations.

Long-term outcome after stifle arthroscopy in 82 Western performance horses (2003-2010).

OBJECTIVE: To report the outcome of horses engaged in Western performance disciplines after stifle arthroscopy and identify prognostic factors for return to performance. STUDY DESIGN: Retrospective case series. SAMPLE POPULATION: Eighty-two Western performance horses undergoing stifle arthroscopy. METHODS: Medical records were reviewed for horses involved in athletic performance/training for various Western performance disciplines and undergoing arthroscopy for lameness localized to the stifle. Follow-up was obtained ≥2 years postoperatively by telephone interviews with the owners. Preoperative and intraoperative findings as well as postoperative treatment were analyzed for their association with return to athletic performance as the primary outcome of interest. RESULTS: The most common disciplines represented were cutting (n = 38), Western pleasure (n = 13), and reining (n = 13). Approximately 40% (32/82) of horses returned to intended use after surgery. Increased age, higher degree of lameness, longer duration of lameness, and the presence of partial-thickness cartilage lesions decreased the odds of returning to athletic performance. Postoperative therapies (intra-articular: stem cells, corticosteroids, interleukin-1 receptor antagonist protein, hyaluronic acid/polysulfated glycosaminoglycans; systemic: nonsteroid anti-inflammatory drugs, hyaluronic acid/polysulfated glycosaminoglycans, oral joint supplements) did not affect the odds of returning to intended use. CONCLUSION: Less than half of the Western performance horses that underwent stifle arthroscopy returned to intended use. Older age, longer duration of lameness, and presence of partial-thickness cartilage lesions affected the odds of a horse returning to intended use. Postoperative therapies did not affect the outcome in this population. CLINICAL SIGNIFICANCE: The prognosis of Western performance horses undergoing stifle arthroscopy is as guarded as that previously reported in horses of other disciplines.

Gait and electromyographic alterations due to early onset of injury and eventual rupture of the cranial cruciate ligament in dogs: A pilot study.

OBJECTIVE: Identify relevant electromyography (EMG), kinematic, and kinetic changes resulting from monopolar radiofrequency energy (MRFE)-induced cranial cruciate ligament (CCL) injury and eventual rupture in dogs. STUDY DESIGN: Experimental, repeated measures. ANIMALS: Five purpose-bred female dogs free of orthopedic and neurologic disease. METHODS: Surface EMG, joint kinematics, and ground reaction forces were assessed at a trot in the pelvic limbs at baseline, at 2 and 4 weeks after unilateral MRFE-induced CCL injury, and at 4, 8, and 16 weeks after CCL rupture (CCLR). RESULTS: After MRFE-induced injury, average hip joint range of motion (ROM) during stance decreased within the untreated pelvic limb. After CCLR, stifle flexion angles decreased within the treated limb at 8 weeks and within the untreated pelvic limb at all time points, whereas average tarsal joint ROM decreased in the treated limb and increased in the untreated limb. Peak vertical ground reaction force and impulse decreased within the treated limb. Qualitative alterations of many EMG values were noted after MRFE-induced injury and CCLR, although significant differences between limbs or from baseline values were not detected. CONCLUSION: Monopolar radiofrequency energy-induced injury altered contralateral hip kinematics, suggesting early regional compensatory gait alterations. After CCLR, additional compensatory gait patterns occurred in both pelvic limbs. CLINICAL IMPACT: The qualitative analysis of trial-averaged EMG data in this small population supports a relationship between neuromuscular function and induced CCL injury leading to rupture.

The Effects of Mepivacaine Hydrochloride on Antimicrobial Activity and Mechanical Nociceptive Threshold During Amikacin Sulfate Regional Limb Perfusion in the Horse.

OBJECTIVE: To determine the effect of intravenous regional limb perfusion (IVRLP) with a combination of mepivacaine hydrochloride and amikacin sulfate on synovial fluid amikacin sulfate concentration, antimicrobial activity, and mechanical nociceptive threshold (MNT). STUDY DESIGN: Experimental study. ANIMALS: Healthy adult horses (n=9). METHODS: One IVRLP treatment was randomly administered by cephalic vein of each limb: amikacin alone (1 g amikacin in 60 mL saline) or amikacin with mepivacaine (1 g amikacin and 500 mg mepivacaine in 60 mL saline). Opposite treatments were repeated after a 24 hour wash-out period. Amikacin concentration and antimicrobial activity were determined for synovial fluid from middle carpal joints at tourniquet removal and 30 minutes following. Zone of inhibition was determined for Staphylococcus aureus and Escherichia coli. MNT was determined at 3 dorsal metacarpal locations prior to and after sedation, after Esmarch tourniquet application, and 30 minutes after IVRLP prior to and after tourniquet removal. RESULTS: Two limbs from each treatment group were removed because of undetectable amikacin concentrations for a total of 14 data sets analyzed. Synovial fluid amikacin concentrations and zone of inhibition were not significantly different between treatments at any time point. MNT were significantly increased 30 minutes after IVRLP prior to and following tourniquet removal using amikacin and mepivacaine (median, range; 40.0 µg/mL, 38.7-40.0 and 40.0, 25.8-40.0, respectively) compared to amikacin alone (19.5 µg/mL, 18.7-25.6 and 15.3, 13.2-20.5, respectively). CONCLUSION: Addition of mepivacaine to amikacin for IVRLP in the horse as a means of providing analgesia without decreasing antimicrobial activity.

COMPARISON BETWEEN COMPUTED TOMOGRAPHIC ARTHROGRAPHY, RADIOGRAPHY, ULTRASONOGRAPHY, AND ARTHROSCOPY FOR THE DIAGNOSIS OF FEMOROTIBIAL JOINT DISEASE IN WESTERN PERFORMANCE HORSES.

The femorotibial joints are a common source of lameness in Western performance horses. The objective of this prospective study was to compare the radiography, ultrasonography, computed tomographic arthrography (CTA), and arthroscopy findings in horses with lameness localized to the femorotibial joints. Twenty-five stifles in 24 horses were included and were evaluated with all four of these diagnostic methods. Defects detected in femorotibial joint structures were compared between diagnostic methods using a McNemar's test to evaluate for disagreement. Cranial medial meniscotibial desmopathy was most detected on arthroscopy (in 14/25 cases) and was only detected on ultrasonography in three out of 11 (27.3%) arthroscopically observed cases, but was detected on CTA in nine out of 12 (75%) arthroscopically observed cases. Medial meniscal injury located on the craniolateral border was most detected on arthroscopy (n = 9) and was detected on CTA in five cases, but on ultrasonography in 0 cases. Detection of articular cartilage defects on the medial femoral condyle was most detected with arthroscopy (24/25, 96% cases) and was also detected on CTA in 12/20 (60%) cases with a significant disagreement identified between modalities (P = 0.02). Cranial and caudal cruciate ligament defects were detected on CTA in 6/22 (27.3%) and 7/19 (36.8%) cases, respectively, and with arthroscopy in 3/25 (12%) and 2/25 (8%) cases, respectively. The use of CTA detected more defects in the cruciate ligaments, proximal tibia, and ligament entheses than the other diagnostic methods, but was not reliable for detection of articular cartilage damage on the medial femoral condyle.

Diagnostic stifle joint arthroscopy using a needle arthroscope in standing horses.

OBJECTIVE: To assess use of an 18 g arthroscope for diagnostic stifle joint examination in the standing horse. STUDY DESIGN: Phase 1 used cadaver limbs and simultaneous ultrasonographic assessment. Phase 2 used 6 normal horses where stifles were assessed in both a standing and flexed position. Phase 3 used horses with suspected stifle injury or disease. ANIMALS: Normal horses (n = 6) to assess ability to perform diagnostic procedure (phase 2) and 3 clinical cases (phase 3). METHODS: Five cadaver limbs were used in phase 1 to assess all stifle joints. Phase 2 used standing sedated and locally anesthetized horses. Routine arthroscopic approaches were used in both weight bearing and flexed nonweight bearing positions. In both phase 1 and 2 simultaneous ultrasonographic and arthroscopic examinations were used to confirm extent of diagnostic examination. The methods developed in phase 2 were used to examine the stifle in 3 horses with suspected stifle disease. RESULTS: In cadaveric limbs and horses, all intra articular structures that constitute a complete arthroscopic examination were identified; no intra -or postoperative morbidity occurred. In phase 3, the needle arthroscope was used in accurate identification of pathologic change and in 1 horse, an osteochondral fragment not detected by ultrasonography and radiography was identified. CONCLUSIONS: This preliminary work indicates that an 18 g arthroscope can be used for diagnostic examination of the equine stifle in standing horses.

Treatment of experimentally induced osteoarthritis in horses using an intravenous combination of sodium pentosan polysulfate, N-acetyl glucosamine, and sodium hyaluronan.

OBJECTIVE: To assess the effects of sodium pentosan polysulfate (PPS), N-acetyl glucosamine (NAG), and sodium hyaluronan (HA) in horses with induced osteoarthritis (OA). STUDY DESIGN: Experimental. ANIMALS: Adult Standard bred horses (n = 16). METHODS: OA was induced arthroscopically in 1 intercarpal joint; 8 horses were administered 3 mg/kg PPS, 4.8 mg/kg NAG, and 0.12 mg/kg HA (PGH), intravenously (IV), weekly and 8 horses were administered an equivalent volume of saline IV until study completion (day 70). Horses underwent a standardized treadmill exercise program. Clinical and radiographic findings and synovial fluid analysis were evaluated throughout the study. Macroscopic, histologic, histochemical, and biochemical findings were evaluated after necropsy. Comparisons of interest included OA and non-OA joints of saline treated horses and OA joints of PGH treated horses and OA joints of saline treated horses. Results were statistically analyzed with significance set at P < .05. RESULTS: OA caused increases in clinical assessment scores, synovial fluid variables, radiographic, macroscopic, and histologic cartilage scores, synovial fluid and cartilage chondroitin sulfate 846-epitope and glycosaminoglycan concentration. Total radiographic scores, total macroscopic joint pathology and macroscopic cartilage pathology scores were significantly reduced in horses treated with PGH compared with saline treated horses. Synovial fluid total protein concentration and white blood cell count were higher in OA joints of PGH treated horses compared with saline treated horses. There were no other significant differences between treatment groups. CONCLUSIONS: Improvements in macroscopic variables were not supported by other outcomes. Further evidence is needed before PGH can be recommended as a therapeutic option for osteoarthritis in horses.

Clinical outcome after intra-articular administration of bone marrow derived mesenchymal stem cells in 33 horses with stifle injury.

OBJECTIVE: To report outcome of horses with femorotibial lesions (meniscal, cartilage or ligamentous) treated with surgery and intra-articular administration of autologous bone marrow derived mesenchymal stem cells (BMSCs). STUDY DESIGN: Prospective case series. ANIMALS: Horses (n = 33). METHODS: Inclusion criteria included horses that had lameness localized to the stifle by diagnostic anesthesia, exploratory stifle arthroscopy and subsequent intra-articular administration of autologous BMSCs. Case details and follow-up were gathered from medical records, owner, trainer or veterinarian. Outcome was defined as returned to previous level of work, returned to work, or failed to return to work. RESULTS: Follow-up (mean, 24 months) was obtained; 43% of horses returned to previous level of work, 33% returned to work, and 24% failed to return to work. In horses with meniscal damage (n = 24) a higher percentage in the current study (75%) returned to some level of work compared to those in previous reports (60-63%) that were treated with arthroscopy alone, which resulted in a statistically significant difference between studies (P = .038). Joint flare post injection was reported in 3 horses (9.0%); however, no long-term effects were noted. CONCLUSIONS: Intra-articular administration of BMSC postoperatively for stifle lesions appeared to be safe, with morbidity being similar to that of other biologic agents. Improvement in ability to return to work may be realized with BMSC treatment compared to surgery alone in horses with stifle injury.

Intra-articular corticosteroids for the treatment of osteoarthritis: A systematic review and meta-analysis on the comparison of different molecules and doses.

Purpose: The purpose of this study was to quantify and compare the clinical relevance of the different intra-articular corticosteroids (CS) effects in vivo for osteoarthritis (OA) treatment. Methods: The search was conducted on PubMed, Cochrane, and Web of Science in October 2023. The PRISMA guidelines were used. Inclusion criteria: animal or human randomized controlled trials (RCTs), English language and no time limitation, on the comparison of different intra-articular CS for OA treatment. The articles' quality was assessed using the Cochrane RoB2 and GRADE guidelines for human RCTs, and SYRCLE's tool for animal RCTs. Results: Eighteen RCTs were selected (16 human and 2 animal studies), including 1577 patients (1837 joints) and 31 animals (51 joints). The CS used were triamcinolone (14 human and 2 animal studies), methylprednisolone (7 human and 1 animal study), betamethasone (3 human studies) and dexamethasone (1 human study). All studies addressed knee OA except for three human and one animal study. A meta-analysis was performed on the comparison of methylprednisolone and triamcinolone in humans with knee OA analysing VAS pain at very short- (≤2 weeks), short- (>2 and ≤4 weeks), mid- (>4 and ≤8 weeks), long- (>8 and ≤ 12 weeks), and very long-term (>12 and ≤24 weeks). Triamcinolone showed better post-injection values compared to methylprednisolone at very short-term (p = 0.028). No difference in terms of VAS improvement was observed at any follow-up. Conclusions: The available preclinical and clinical literature provides limited evidence on the comparison of different CS, hindering the possibility of determining the best CS approach in terms of molecule and dose for the intra-articular injection of OA joints. Level of Evidence: Level I.

Infrared Spectroscopy of Synovial Fluid Shows Accuracy as an Early Biomarker in an Equine Model of Traumatic Osteoarthritis.

Osteoarthritis is a leading cause of lameness and joint disease in horses. A simple, economical, and accurate diagnostic test is required for routine screening for OA. This study aimed to evaluate infrared (IR)-based synovial fluid biomarker profiling to detect early changes associated with a traumatically induced model of equine carpal osteoarthritis (OA). Unilateral carpal OA was induced arthroscopically in 9 of 17 healthy thoroughbred fillies; the remainder served as Sham-operated controls. The median age of both groups was 2 years. Synovial fluid (SF) was obtained before surgical induction of OA (Day 0) and weekly until Day 63. IR absorbance spectra were acquired from dried SF films. Following spectral pre-processing, predictive models using random forests were used to differentiate OA, Sham, and Control samples. The accuracy for distinguishing between OA and any other joint group was 80%. The classification accuracy by sampling day was 87%. For paired classification tasks, the accuracies by joint were 75% for OA vs. OA Control and 70% for OA vs. Sham. The accuracy for separating horses by group (OA vs. Sham) was 68%. In conclusion, SF IR spectroscopy accurately discriminates traumatically induced OA joints from controls.

IL-1ra gene therapy in equine osteoarthritis improves physiological, anatomical, and biological outcomes of joint degeneration.

OBJECTIVE: To evaluate the effects of a gene transfer approach to IL-1ß inhibition in an equine osteochondral chip fragment model of joint injury using a self-complementary adeno-associated virus with interleukin receptor antagonist transgene cassette (scAAVIL-1ra), as posttraumatic osteoarthritis in horses, similar to people, is a significant clinical problem. ANIMALS: 16 horses were utilized for the study. METHODS: All horses had an osteochondral chip fragment induced arthroscopically in one middle carpal joint while the contralateral joint was sham operated. Eight horses received either scAAVIL-1ra or saline in the osteoarthritis joint. Horses were evaluated over 70 days clinically (lameness, imaging, and biomarker analysis) and euthanized at 70 days and evaluated grossly, with imaging and histopathology. RESULTS: The following findings were statistically significant. Injection of scAAVIL-1ra resulted in high synovial fluid levels of IL-1ra (0.5 to 9 µg/mL) throughout the duration of the experiment (70 days). Over the duration, we observed scAAVIL-1ra to improve lameness (lameness score relative improvement of 1.2 on a scale of 0 to 5), cause suppression of prostaglandin E2 (a relative decline of 30 pg/mL), and result in histological improvement in articular cartilage (decreased chondrocyte loss and chondrone formation) and subchondral bone (less osteochondral splitting and osteochondral lesions). Within the synovial membrane of scAAVIL-1ra-treated joints, we also observed perivascular infiltration with CD3-positive WBCs, suggesting lymphocytic T-cell perivascular infiltration commonly observed with viral transduction. CLINICAL RELEVANCE: These data provide support for further evaluation and optimization of scAAVIL-1ra gene therapy to treat equine osteoarthritis.

Radiological findings in the proximal sesamoid bones of yearling and 2-year-old Thoroughbred sales horses: Prevalence, progression and associations with racing performance.

BACKGROUND: Radiological findings in the proximal sesamoid bones (sesamoids) are a persistent source of controversy at Thoroughbred sales, due to inconsistent classification and conflicting assignment of potential clinical importance. OBJECTIVES: To define the prevalence of sesamoid findings on sales repository radiographs in yearling and 2-year-old Thoroughbreds and to analyse associations with racing performance. To track the changes in sesamoid findings between 1 and 2 years of age in horses that present for sale at both ages. STUDY DESIGN: Prospective cohort study using an enrolled sample. METHODS: Horses were enrolled from a 2016 yearling sale and five 2017 2-year-old sales with consignor permission. Radiological findings relating to sesamoid vascular channel appearance, abaxial contour changes and sesamoid fragments were examined. Associations between sesamoid findings and racing performance from 2 to 4 years of age were examined using multivariate regression analyses. Clinical follow-up was sought to ascertain why horses that did not race never started. RESULTS: A total of 2508 yearlings and 436 2-year-olds were included for evaluation. Interobserver agreement using the new grading system was substantial. Yearling findings associated with a significantly reduced probability of starting a race were: Grade 3 vascular channels in forelimb sesamoids (0.52, P < 0.001, 95% confidence interval (CI): 0.37-0.67), abaxial new bone in forelimb sesamoids (0.62, P = 0.01, 95% CI: 0.49-0.73), apical or abaxial fragments in forelimb sesamoids (0.55, P = 0.005, 95% CI: 0.37-0.72). For affected horses that did race, Grade 3 vascular channels in forelimb sesamoids were associated with fewer race starts (9.9 starts, P = 0.03, 95% CI: 8.0-12.2) and Grade 3 vascular channels in hindlimb sesamoids were associated with a delayed start to racing careers (54 days, P = 0.01, 95% CI: 20-89). Abaxial new bone in forelimb sesamoids was associated with a 54% reduction in total earnings (P = 0.003, 95% CI: 24-72) and a 46% reduction in earnings per start (P = 0.002, 95% CI: 21-64). Abaxial concavity occurred predominantly in yearling medial forelimb sesamoids, had no impact on racing performance and mostly resolved by 2-year-old sale. MAIN LIMITATIONS: These findings are applicable to horses presented for sale at public auction and may underestimate the prevalence of severe lesions in non-sales horses. CONCLUSIONS: Grade 3 vascular channels, forelimb sesamoid abaxial new bone and forelimb sesamoid fragments are important findings in sales repository radiology. The new grading scale assigns a numerical grade for vascular channel appearance that matches the number of enlarged vascular channels evident in a given sesamoid. Abaxial contour changes, when present in sesamoids that are Grade 0 for enlarged vascular channels, are noted separately as either abaxial new bone or abaxial concavity. Fragments are also noted and interpreted separately.

Longitudinal in vivo cationic contrast-enhanced computed tomography classifies equine articular cartilage injury and repair.

Cationic contrast-enhanced computed tomography (CECT) capitalizes on increased contrast agent affinity to the charged proteoglycans in articular cartilage matrix to provide quantitative assessment of proteoglycan content with enhanced images. While high resolution microCT has demonstrated success, we investigate cationic CECT use in longitudinal in vivo imaging at clinical resolution. We hypothesize that repeated administration of CA4+ will have no adverse side effects or complications, and that sequential in vivo imaging assessments will distinguish articular cartilage repair tissue from early degenerative and healthy cartilage in critically sized chondral defects. In an established equine translational preclinical model, lameness and synovial effusion scores are similar to controls after repeated injections of CA4+ (eight injections over 16 weeks) compared to controls. Synovial fluid total protein, leukocyte concentration, and sGAG and PGE2 concentrations and articular cartilage and synovial membrane scores are also equivalent to controls. Longitudinal in vivo cationic CECT attenuation in repair tissue is significantly lower than peripheral to (adjacent) and distantly from defects (remote sites) by 4 weeks (p < 0.001), and this difference persists until 16 weeks. At the 6- and 8-week time points, the adjacent locations exhibit significantly lower cationic CECT attenuation compared with the remote sites, reflecting peri-defect degeneration (p < 0.01). Cationic CECT attenuation at clinical resolution significantly correlates with cationic CECT (microCT) (r = 0.69, p < 0.0001), sGAG (r = 0.48, p < 0.0001), and ICRS II histology score (r = 0.63, p < 0.0001). In vivo cationic CECT imaging at clinical resolution distinguishes fibrous repair tissue from degenerative and healthy hyaline cartilage and correlates with molecular tissue properties of articular cartilage.

Effects of equine joint injury on boundary lubrication of articular cartilage by synovial fluid: role of hyaluronan.

OBJECTIVE: To compare equine synovial fluid (SF) from injured and control joints for cartilage boundary lubrication function; concentrations of the putative boundary lubricant molecules hyaluronan (HA), proteoglycan 4 (PRG4), and surface-active phospholipids (SAPLs); relationships between lubrication function and composition; and lubrication restoration by addition of HA. METHODS: Equine SF from normal joints, joints with acute injury, and joints with chronic injury were analyzed for boundary lubrication of normal articular cartilage (kinetic friction coefficient [µ(kinetic) ]). Equine SF samples were analyzed for HA, PRG4, and SAPL concentrations and HA molecular weight distribution. The effect of the addition of HA, of different concentrations and molecular weight, on the µ(kinetic) of equine SF samples from normal joints and joints with acute injury was determined. RESULTS: The µ(kinetic) of equine SF from joints with acute injury (0.036) was higher (+39%) than that of equine SF from normal joints (0.026). Compared to normal equine SF, SF from joints with acute injury had a lower HA concentration (-30%) of lower molecular weight forms, higher PRG4 concentration (+83%), and higher SAPL concentration (+144%). Equine SF from joints with chronic injury had µ(kinetic) , PRG4, and SAPL characteristics intermediate to those of equine SF from joints with acute injury and normal equine SF. Regression analysis revealed that the µ(kinetic) value decreased with increasing HA concentration in equine SF. The friction-reducing properties of HA alone improved with increasing concentration and molecular weight. The addition of high molecular weight HA (4,000 kd) to equine SF from joints with acute injury reduced the µ(kinetic) to a value near that of normal equine SF. CONCLUSION: In the acute postinjury stage, equine SF exhibits poor boundary lubrication properties, as indicated by a high µ(kinetic) . HA of diminished concentration and molecular weight may be the basis for this, and adding HA to deficient equine SF restored lubrication function.

Synoviocytes protect cartilage from the effects of injury in vitro.

BACKGROUND: It is well documented that osteoarthritis (OA) can develop following traumatic joint injury and is the leading cause of lameness and subsequent wastage of equine athletes. Although much research of injury induced OA has focused on cartilage, OA is a disease that affects the whole joint organ. METHODS: In this study, we investigated the impact of synovial cells on the progression of an OA phenotype in injured articular cartilage. Injured and control cartilage were cultured in the presence of synoviocytes extracted from normal equine synovium. Synoviocytes and cartilage were evaluated for catabolic and anabolic gene expression. The cartilage was also evaluated histologically for loss of extracellular matrix molecules, chondrocyte cell death and chondrocyte cluster formation. RESULTS: The results indicate synoviocytes exert both positive and negative effects on injured cartilage, but ultimately protect injured cartilage from progressing toward an OA phenotype. Synoviocytes cultured in the presence of injured cartilage had significantly reduced expression of aggrecanase 1 and 2 (ADAMTS4 and 5), but also had increased expression of matrix metalloproteinase (MMP) -1 and reduced expression of tissue inhibitor of metalloproteinases 1 (TIMP-1). Injured cartilage cultured with synoviocytes had increased expression of both collagen type 2 and aggrecanase 2. Histologic examination of cartilage indicated that there was a protective effect of synoviocytes on injured cartilage by reducing the incidence of both focal cell loss and chondrocyte cluster formation, two major hallmarks of OA. CONCLUSIONS: These results support the importance of evaluating more than one synovial joint tissue when investigating injury induced OA.

The role of muscle activation in cruciate disease.

Traditional investigations into the etiopathogenesis of canine cranial cruciate ligament (CCL) disease have focused primarily on the biological and mechanical insults to the CCL as a passive stabilizing structure of the stifle. However, with recent collaboration between veterinarians and physical therapists, an increased focus on the role of muscle activity and aberrant motor control mechanisms associated with anterior cruciate ligament (ACL) injuries and rehabilitation in people has been transferred and applied to dogs with CCL disease. Motor control mechanisms in both intact and cruciate-deficient human knees may have direct translation to canine patients, because the sensory and motor components are similar, despite moderate anatomic and biomechanical differences. Components of motor control, such as muscle recruitment and the coordination and amplitudes of activation are strongly influenced by afferent proprioceptive signaling from peri- and intra-articular structures, including the cruciate ligaments. In people, alterations in the timing or amplitude of muscle contractions contribute to uncoordinated movement, which can play a critical role in ACL injury, joint instability and the progression of osteoarthritis (OA). A better understanding of motor control mechanisms as they relate to canine CCL disease is vitally important in identifying modifiable risk factors and applying preventative measures, for development of improved surgical and rehabilitative treatment strategies. The purpose of this review article is to analyze the influence of altered motor control, specifically pelvic limb muscle activation, in dogs with CCL disease as evidenced by mechanisms of ACL injury and rehabilitation in people.

Plasma and Synovial Fluid Cell-Free DNA Concentrations Following Induction of Osteoarthritis in Horses.

Biomarkers for osteoarthritis (OA) in horses have been extensively investigated, but translation into clinical use has been limited due to cost, limited sensitivity, and practicality. Identifying novel biomarkers that overcome these limitations could facilitate early diagnosis and therapy. This study aimed to compare the concentrations of synovial fluid (SF) and plasma cell-free DNA (cfDNA) over time in control horses with those with induced carpal OA. Following an established model, unilateral carpal OA was induced in 9 of 17 healthy Thoroughbred fillies, while the remainder were sham-operated controls. Synovial fluid and plasma samples were obtained before induction of OA (Day 0) and weekly thereafter until Day 63, and cfDNA concentrations were determined using fluorometry. The SF cfDNA concentrations were significantly higher for OA joints than for sham-operated joints on Days 28 (median 1430 µg/L and 631 µg/L, respectively, p = 0.017) and 63 (median 1537 µg/L and 606 µg/L, respectively, p = 0.021). There were no significant differences in plasma cfDNA between the OA and the sham groups after induction of carpal OA. Plasma cfDNA measurement is not sufficiently sensitive for diagnostic purposes in this induced model of OA. Synovial fluid cfDNA measurement may be used as a biomarker to monitor early disease progression in horses with OA.