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.

A safety evaluation of allogeneic freeze-dried platelet-rich plasma or conditioned serum compared to autologous frozen products equivalents in equine healthy joints.

BACKGROUND: Hemoderivatives such as autologous conditioned serum (ACS) and platelet-rich plasma (PRP) have been used as potential disease-modifying therapies in musculoskeletal disorders such as osteoarthritis (OA). These therapies are based on the delivery of multiple growth factors and anti-inflammatory cytokines that are known to participate in inflammatory processes. The variability of cytokine content due to the autologous nature of the product, the non-availability for immediate use and need for storage at low temperatures are limitations for its use in the field. An allogeneic freeze-dried conditioned serum (CS) and PRP would provide field clinicians with a more practical approach to use such products in daily practice. Based on in vitro preliminary data, this experimental study aimed to test the in vivo safety of allogeneic freeze-dried CS and PRP in healthy joints, using the horse as a model. RESULTS: Eight horses were randomly assigned and treated with PRP or CS. Horses had three joints injected with ALLO-FD PRP or CS, and three contralateral joints injected with the AUTO version of the same product, by a blinded clinician. Horses were evaluated clinically, and had synovial fluid collected at different time points and evaluated for cell content, PGE2 and protein. Both CS and PRP products triggered a self-limiting and mild inflammatory response in equine healthy joints. This was indicated by the transient increase in nucleated cell count, PGE2 and total protein in synovial fluid. This mild inflammatory response did not result in significant lameness and was not different among the groups. CONCLUSIONS: The allogeneic freeze-dried PRP and CS showed to be overall safe and not dissimilar compared to their autologous frozen version in equine healthy joints. Further studies are necessary to evaluate the modulatory effects of these therapies in a clinical setting.

Tissue predictability of elastography is low in collagenase induced deep digital flexor tendinopathy.

Elastography is an emerging imaging modality for characterizing tendon injury in horses, but its ability to differentiate tissue deformability relative to treatment group and biochemical properties using a prospective, experimental study design remain unknown. Objectives of the current study were to (a) to investigate differences in glycosaminoglycan, DNA, and soluble collagen levels in mesenchymal stem cell (MSC) treated limbs compared to untreated control limbs utilizing a collagenase model of tendinopathy; (b) compare elastographic features between treatment groups; and (c) determine tissue-level predictive capabilities of elastography in relation to biochemical outcomes. Bone marrow was collected for MSC culture and expansion. Tendinopathy of both forelimb deep digital flexor tendons (DDFTs) was induced with collagenase under ultrasonographic guidance. One randomly assigned limb was treated with intra-lesional MSC injection with the opposite limb serving as an untreated control. Horses were placed into a controlled exercise program with elastographic evaluations performed baseline (0) and 14, 60, 90, and 214 days post-treatment. Postmortem biochemical analysis was performed. MSC-treated limbs demonstrated significantly less (42%) glycosaminoglycan (P = .006). Significant differences in elastographic region of interest (ROI) percent hardness, ROI color histogram, and subjective lesion stiffness were appreciated between treatment groups at various study time points. Elastographic outcome parameters were weak predictors of biochemical tissue analysis, with all R2 values ≤ 0.50. Within this range of differences in glycosaminoglycan content between treatment groups, elastography outcomes did not predict biochemical differences. Tissue-specific differences between DDFTs treated with MSCs compared to controls were apparent biochemically, but not predicted by elastography.

Revisiting predictive biomarkers of musculoskeletal injury in thoroughbred racehorses: longitudinal study in polish population.

BACKGROUND: High prevalence of musculoskeletal disorders in racehorses and its impact on horse welfare and racing economics call for improved measures of injury diagnosis and prevention. Serum biomarkers of bone and cartilage metabolism have previously shown promise in prediction of musculoskeletal injuries in horses. This study aimed to re-evaluate usability of the predictive serum biomarkers identified in North American Thoroughbred racehorses in a geographically distinct group of Polish Thoroughbreds. RESULTS: Serum concentrations of bone and cartilage biomarkers: osteocalcin, c-terminal telopeptide of type I collagen, total glycosaminoglycans (GAG), chondroitin sulfate epitope and c-propeptide of type II procollagen (CPII) were evaluated in the beginning and the next 3 months of one racing season in a cohort of twenty-six 2-year-old Polish racehorses. Exit criteria were diagnosis of musculoskeletal injury, leading to > 5 days off training (n = 8), or completion of 3 study months with no training interruptions (n = 18). Normalized results and matching archival data from 35 2-year-old North American racehorses was used for logistic regression analysis to identify universal predictors of injury. Mean GAG and CPII levels were lower in injured group comparing to control, which is consistent with previous findings in racehorses. These biomarkers were also identified as predictors of injury in the mixed population model. Population origin had no significant effect on predictive value of evaluated biomarkers (Wald test p = 0.137). Decreased osteocalcin and increased c-terminal telopeptide of type I collagen levels in injured horses comparing to controls were specific for Polish population and signalized disruption in bone turnover homeostasis. CONCLUSIONS: Changes in serum GAG and CPII in racehorses at risk of injury appear to be similar across distinct populations while dynamics of serum bone marker is more population-specific.

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.

High field magnetic resonance imaging is comparable with gross anatomy for description of the normal appearance of soft tissues in the equine stifle.

High field magnetic resonance imaging (MRI) is increasingly used for horses with suspected stifle disease, however there is limited available information on normal imaging anatomy and potential incidental findings. The aim of this prospective, anatomic study was to develop an optimized high field MRI protocol for evaluation of the equine stifle and provide detailed descriptions of the normal MRI appearance of the stifle soft tissues, using ultrasound and gross pathological examination as comparison tests. Nine cadaver limbs were acquired from clinically normal horses. Stifles were evaluated ultrasonographically and then by an extensive 1.5 T MRI protocol. Subsequently, all stifles were evaluated for gross pathologic change. Findings were compared between gross evaluation and MRI imaging and described. No soft tissue structure abnormalities were identified on any evaluation. Specific descriptive findings of the meniscotibial, meniscofemoral, collateral, patellar and cruciate ligaments, and the menisci were reported. The high field MRI protocol described in this study provided high spatial and contrast resolution of the soft tissue structures, and this in turn allowed visualization of detailed structural characteristics, such as striations and variations in signal intensity. Findings supported the use of high field MRI as a modality for the evaluation of the soft tissues of the equine stifle. As clinical availability of this modality increases in the future, authors anticipate that new stifle diseases will be detected that have not previously been identified with other imaging modalities.

Additional palmaroproximal-palmarodistal oblique radiographic projections improve accuracy of detection and characterization of equine flexor cortical lysis.

Radiographic flexor cortical lysis indicates advanced degenerative change and its earlier recognition may improve case outcome. Aims of this prospective, diagnostic accuracy study were to determine effects of radiographic beam angle and observer on accuracy of lesion detection. The sample included 36 limbs from 31 horses. Palmaroproximal-palmarodistal oblique (skyline) radiographs were acquired at standard (n = 38) and more shallow (alternate) angles (n = 58). Images were independently reviewed by four experience levels of five observers each (n = 20) for the presence and severity of flexor cortical lysis. Observers also reported their confidence in these answers. Responses were compared based on seeing a standard skyline or multiple projections. The definitive presence (or absence) and severity of lysis was based upon radiologist consensus agreement. When assessed by observer, the identification of lysis and the assessment of its severity was most similar to that of radiologists when observers of all levels of experience were able to view multiple skyline projections (P = 0.399, P = 0.174). Using multiple views to detect lysis resulted in improved sensitivity (85.3% vs. 97.2%, P < 0.001), decreased specificity (82.8% vs. 74.5%, P = 0.03), and improved interobserver agreement (86.0% vs. 90.2%, P = 0.21). On average, observers of all levels of experience became more confident viewing multiple projections (P < 0.001). Skyline radiographs using a flatter angle of incidence improve radiographic detection and characterization of flexor cortical lysis severity, may allow the beam to be tangential to the more distal portion of the navicular bone, and are recommended when flexor cortical lysis is suspected.

High field magnetic resonance imaging contributes to diagnosis of equine distal tarsus and proximal metatarsus lesions: 103 horses.

Injury of the distal tarsus and proximal metatarsus commonly causes lameness. Magnetic resonance imaging (MRI) allows concurrent assessment of both the distal tarsal joints and suspensory ligament origin, and aids identification of lesions that may otherwise go undetected by other modalities. In this retrospective observational study, the medical records of a veterinary imaging center were searched for MRI exams of the distal tarsus and proximal metatarsus for the years 2012 through 2014. Studies for 125 limbs of 103 horses were identified and retrospectively evaluated by two board-certified veterinary radiologists. Soft tissue and osseous changes were characterized and graded by degree of severity. The patients' signalment, lameness severity, and results of diagnostic analgesia were recorded. Osteoarthritic changes of the distal intertarsal and tarsometatarsal joints were the most common findings. Other findings included bone marrow lesions, degenerative changes of the small cuboidal bones, subchondral cystic lesions, and intertarsal desmopathy. Suspensory ligament desmopathy was found in 53% of limbs. Fourty-seven percent of limbs that responded to analgesia of the proximal suspensory ligament had more severe lesions in the distal tarsus. Bone marrow lesions of the third tarsal bone were the only MRI finding that correlated with grade of lameness in patients for which lameness grade was reported. The grade of lameness has a poor correlation with the severity of lesions found on MRI. The findings support the use of MRI for simultaneous evaluation of the proximal metatarsus and distal tarsus, particularly given the difficulty of lesion localization with diagnostic analgesia.

Prospective In Vivo Comparison of Damaged and Healthy-Appearing Articular Cartilage Specimens in Patients With Femoroacetabular Impingement: Comparison of T2 Mapping, Histologic Endpoints, and Arthroscopic Grading.

PURPOSE: To describe T2 mapping values in arthroscopically determined International Cartilage Repair Society (ICRS) grades in damaged and healthy-appearing articular cartilage waste specimens from arthroscopic femoroacetabular impingement (FAI) treatment. Furthermore, we sought to compare ICRS grades of the specimens with biochemical, immunohistochemistry and histologic endpoints and assess correlations with T2 mapping. METHODS: Twenty-four patients were prospectively enrolled, consecutively, between December 2011 and August 2012. Patients were included if they were aged 18 years or older and met criteria that followed the clinical indications for arthroscopy to treat FAI. Patients with prior hip trauma including fracture or dislocation or who have undergone prior hip surgery were excluded. All patients received a preoperative sagittal T2 mapping scan of the hip joint. Cartilage was graded intraoperatively using the ICRS grading system, and graded specimens were collected as cartilage waste for histologic, biochemical, and immunohistochemistry analysis. RESULTS: Forty-four cartilage specimens (22 healthy-appearing, 22 damaged) were analyzed. Median T2 values were significantly higher among damaged specimens (55.7 ± 14.9 ms) than healthy-appearing specimens (49.3 ± 12.3 ms; P = .043), which was most exaggerated among mild (grade 1 or 2) defects where the damaged specimens (58.1 ± 16.4 ms) were significantly higher than their paired healthy-appearing specimens (48.7 ± 15.4 ms; P = .026). Severely damaged specimens (grade 3 or 4) had significantly lower cumulative H&E than their paired healthy-appearing counterparts (P = .02) but was not statistically significant among damaged specimens with mild (grade 1 or 2) defects (P = .198). Among healthy-appearing specimens, median T2 and the percentage of collagen fibers oriented parallel were significantly correlated (rho = 0.425, P = .048). CONCLUSIONS: This study outlines the potential for T2 mapping to identify early cartilage degeneration in patients undergoing arthroscopy to treat FAI. Findings in ICRS grade 1 and 2 degeneration corresponded to an increase in T2 values. Further biochemical evaluation revealed a significant difference between healthy-appearing cartilage and late degeneration in cumulative H&E as well as significantly lower percentage of collagen fibers oriented parallel and a higher percentage of collagen fibers oriented randomly when considering all grades of cartilage damage. LEVEL OF EVIDENCE: Level II, prospective comparative study.

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.

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.

Substantial variability exists in the interpretation of survey radiographs among equine veterinarians.

BACKGROUND: Radiographs are a common diagnostic tool utilised during pre-purchase examinations, yet differences surround their interpretation among equine veterinarians. OBJECTIVES: (1) To determine veterinarians' subjective impressions of a spectrum of radiographic abnormalities regarding suitability for purchase; (2) To determine how veterinarians' years in practice, primary discipline or board certification status influence their subjective impression of radiographic abnormalities, and (3) To determine how horses' discipline, current level of work (working at intended level vs. future prospect) and buyer intent (sell vs. retain ownership) influence veterinarians' subsequent recommendation. STUDY DESIGN: Worldwide electronic survey of veterinary professionals. METHODS: Four radiographic series of navicular bones, tarsi, and stifles, representing varying degrees of pathological change (normal, mild, moderate and marked) were presented. Respondents (n = 211) graded their level of concern from 1 (no concern) to 10 (very concerned) based on the radiographs and case scenarios presented. RESULTS: Overall, as the grade of pathological change increased for each anatomical location, so did the level of veterinarian concern, but a wide range of answers were submitted for all grades of pathology. Board certified practitioners had a higher level of concern over marked radiographic changes of the navicular bone for both western performance and English sport horses as well as prospect horses (mean = 7.2 vs. 6.1; P = 0.009; 95% CI for difference = [0.3, 2.0]). Board certified respondents had a lower level of concern over marked radiographic changes of the tarsus regardless of the performance status of the horse (mean = 7.1 vs. 8.0; P = 0.008; 95% CI for difference = [-1.5, -0.2]). Western performance horse practitioners had a lower level of concern over mild radiographic changes of the tarsus regardless of the performance status of the horse (mean = 3.3 vs. 4.2; P = 0.009; 95% CI for difference = [-1.6, -0.2]). MAIN LIMITATIONS: Only one example of each pathological grade per anatomical region was presented. CONCLUSIONS: On survey radiographs, the perceived clinical relevance of radiographic abnormalities among equine practitioners differs widely, making purchase recommendations highly variable.

Blood flow restriction training does not negatively alter the mechanical strength or histomorphology of uninjured equine superficial digital flexor tendons.

BACKGROUND: Low load exercise training with blood flow restriction (BFR) has become increasingly used by human physical therapists to prescribe controlled exercise following orthopaedic injury; its effects on the equine superficial digital flexor tendon (SDFT), however, are unknown. OBJECTIVE: To investigate outcomes of pressure specific BFR walking exercise on uninjured equine SDFT biomechanics and histomorphology. STUDY DESIGN: Controlled in vivo experiment. METHODS: Four forelimbs of four horses were exposed to 40 BFR-walk sessions (10-min interval walking) on a treadmill over a 56-day study period with their contralateral forelimbs serving as untreated controls. Similarly, four forelimbs of four control horses were exposed to 40 sham cuff walk sessions. On study Day 56, all horses (n = 8) were humanely euthanised and forelimb SDFTs underwent non-destructive biomechanical testing and corresponding histomorphological analysis. Significance in biomechanical parameters between treatment groups was analysed using a mixed-effects ANOVA with Tukey's post-hoc tests. RESULTS: Statistically significant differences in SDFT stiffness for both first (p = 0.02) and last cycles (p = 0.03) were appreciated within the BFR treated group only, with BFR exposed forelimbs being significantly stiffer than the contralateral unexposed forelimbs. When normalised to cross-sectional area, no significant differences were appreciated among treatment groups in elastic modulus for the first (p = 0.5) or last cycles (p = 0.4). No histological differences were appreciated among treatment groups according to Bonar, Movin, or musculotendinous junction evaluation criteria. MAIN LIMITATIONS: Short-term comparisons were performed in a small sample population without correlation to performance outcome measures. Optimal occlusion percentages and walk protocols remain unknown. CONCLUSIONS: This study demonstrated no negative impact of BFR on mechanical strength of the equine SDFT; however, evidence suggests that BFR results in increased tendon stiffness based on biomechanical testing and subsequent calculations. No consistent detrimental histomorphological changes were seen.

CONTEXTO: Exercício de baixa carga com restrição do fluxo sanguíneo (RFS) tem sido cada vez mais utilizado por fisioterapeutas humanos para tratar lesões ortopédicas. Porém, seus efeitos no tendão flexor digital superficial (TFDS) de equinos não é conhecida. OBJETIVOS: O objetivo deste estudo foi investigar o efeito de específicas pressões com RFS durante o passo em cavalos sem lesão no TFDS, por meio de histologia e análise biomecânica. DELINEAMENTO DO ESTUDO: Estudo controlado. MÉTODOS: Quatro membros torácicos de quatro cavalos foram expostos a 40 sessões de RFS durante o passo (10 minutos de caminhada intervalada), ao longo de 56 dias. O membro contralateral foi utilizado como controle. Da mesma forma, quatro membros de quatro cavalos controle foram expostos a 40 sessões simuladas de caminhada com torniquete. No dia 56, todos os cavalos (n = 8) foram eutanasiados, e os TFDS foram submetidos a testes biomecânicos não destrutivos e análise histológica. A significância dos parâmetros biomecânicos entre tratamentos foi analisada utilizando ANOVA de efeitos mistos, seguida pelo teste de Tukey. RESULTADOS: A rigidez do TFDS foi estatisticamente diferente nos primeiros (p = 0.02) e últimos (p = 0.03) ciclos no grupo submetido à RFS, sendo os membros tratados significativamente mais rígidos do que os membros contralaterais não expostos ao tratamento. Quando normalizado para a área transversal, não foi observada diferença significativa entre os grupos de tratamento no módulo de elasticidade para os primeiros (p = 0.5) e últimos (p = 0.4) ciclos. Não foram identificadas diferenças histológicas nos diferentes tipos de tratamento, de acordo com os critérios de avaliação Bonar, Movin e de junção musculo­tendínea. PRINCIPAIS LIMITAÇÕES: Comparações de curto prazo foram realizadas em uma amostra pequena da população, sem correlação com medidas de resultados de desempenho. As porcentagens ideais de oclusão e os protocolos de caminhada permanecem desconhecidos. CONCLUSÕES: Este estudo não demonstrou impacto negativo do RFS na resistência mecânica do TFDS equino; no entanto, as evidências sugerem que a RFS resulta em aumento da rigidez do tendão com base em testes biomecânicos e cálculos subsequentes. Nenhuma alteração histológica prejudicial consistente foi observada.

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.

Chondrocyte Response to Fresh Autologous Conditioned Serum Versus Freeze-Dried Allogenic Conditioned Serum.

OBJECTIVE: To investigate the cytokine release profile and histological response of human cartilage after exposure to autologous conditioned serum (ACS) and freeze-dried allogenic conditioned serum (FD-CS). DESIGN: Cartilage explants were collected from 6 patients undergoing total knee arthroplasty. ACS and FD-CS were created from patient serum samples. Cartilage samples were divided into 6 groups: (1) untreated control, (2) ACS, (3) FD-CS, (4) untreated interleukin (IL)-1ß (5 ng/ml), (5) IL-1ß + ACS, and (6) IL-1ß + FD-CS. After 12 days, cartilage samples were analyzed with glycosaminoglycan (GAG) concentration normalized to wet weight while comparing cytokine concentrations, and histological scoring. RESULTS: There was a significant decrease in pathology scoring for ACS (P = 0.0368) and FD-CS (P = 0.0368) in the IL-1ß injury groups compared with the untreated IL-1ß insult group. ACS and FD-CS significantly mitigate the IL-1ß induced increase in basic fibroblast growth factor (bFGF) (P = 0.0009 and P = 0.0002, respectively). FD-CS showed a significant decrease in IL-1ß concentration in the presence of IL-1ß insult compared with the untreated IL-1ß group (P < 0.0001). ACS-treated samples had significantly higher concentration of tumor necrosis factor (TNF)-α independent of IL-1ß when compared with samples not treated with biologics (P = 0.0053). CONCLUSIONS: Explanted osteoarthritic cartilage responds favorably and equivalently to treatment with ACS and FD-CS from a histological perspective. Both ACS and FD-CS were able to mitigate the IL-1ß-induced increases in bFGF and FD-CS lowered IL-1ß concentration while increasing interleukin-1 receptor antagonist (IL-1Ra) concentration. Although the cytokine profile of cartilage tissue explants treated with FD-CS appears to be different than that of ACS, this difference does not seem to affect biologic activity of FD-CS.

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.

Comparison of Ultrasound and Magnetic Resonance Imaging for Identifying Soft Tissue Abnormalities in the Palmar Aspect of the Equine Digit.

Damage to the soft tissue structures of the digit is a common source of equine lameness. While magnetic resonance imaging (MRI) allows for the most complete diagnostic imaging of the equine digit, ultrasound is more readily available and less expensive. This prospective diagnostic accuracy study compares ultrasound to MRI for the diagnosis of injuries visible with ultrasound within the digit, including the deep digital flexor tendon (DDFT), collateral sesamoidean ligament (CSL), and navicular bursa. Clinical patients underwent an MRI of the digit and a blinded ultrasound of the digit between the heel bulbs, and results of the two modalities were compared. A total of 70 ultrasound and MRI exams of 45 horses were included. Ultrasound had good sensitivity (85%), moderate specificity (60%), and accuracy of 70% for evaluating the dorsal tearing of the DDFT. Accuracy was lower for navicular bursa effusion (67%), navicular bursa proliferation (61%), and CSL enlargement (61%). Tearing of the DDFT distal to the navicular bone was identified with MRI in 27 limbs, 20 of which also had dorsal damage proximal to the navicular bone identified with ultrasound. Ultrasound evaluation remains a useful screening tool, particularly for the assessment of DDFT tearing proximal to the navicular bone but risks under-diagnosing pathology to the navicular bursa and CSL. Clinically significant concurrent damage to the distal DDFT and other osseous and soft tissues in the hoof capsule is unlikely to be identified without MRI.

Equine blood flow restriction training: Safety validation.

BACKGROUND: Blood flow restriction (BFR) has become a key rehabilitative tool for human orthopaedic conditions. With modernised technology and evolution of clinical application, patient-specific delivery of occlusion percentages is now considered the standard of care in human patients due to improved therapeutic outcomes and minimised safety risks. Safety validation and limb occlusion pressure (LOP) data for horses, however, are lacking. OBJECTIVE: (1) To determine if BFR exposure resulted in forelimb biomechanical gait dysfunction as safety validation and (2) to investigate inter-horse and inter-limb LOP differences. STUDY DESIGN: Controlled in vivo experiment. METHODS: Daily unilateral forelimb BFR was performed in four horses over 56 days. Clinical examinations and objective gait analyses were performed on Days 0, 28 and 56. Daily LOP values were determined by Doppler evaluation to deliver 80% vascular occlusion at a walk. A linear mixed model evaluated for differences in lameness, kinetic and kinematic gait parameters. RESULTS: There were no significant differences in forelimb lameness (range of Grades 0-2 across all forelimbs), kinematic or kinetic gait parameters over time or between BFR-exposed and control (contralateral) limbs (p > 0.05). Clinically apparent complications related to BFR such as thrombosis or dermatitis were not appreciated. Significant differences in mean LOP values between various horses (p < 0.001) and measured left (204.48 mmHg) and right (173.78 mmHg) forelimbs (p < 0.001) were observed. Mean LOP and standard deviation across all readings was 189.1 ± 22.2 mmHg. MAIN LIMITATIONS: Optimal BFR occlusion percentages and protocols with documented clinical efficacy are unknown. Small study population. CONCLUSIONS: Exposure to BFR did not result in forelimb biomechanical dysfunction in four horses. Applied pressures of 75-151 mmHg would likely simulate a range of 50%-80% vascular occlusion in horses, but inherent physiological variation between horses and forelimbs warrants incorporation of individual pressures.

Radiographic changes of the proximal third metatarsal bone do not predict presence or severity of proximal suspensory desmopathy in a predominately Quarter Horse population.

BACKGROUND: Comparison of radiography to magnetic resonance imaging (MRI) can help objectively assess the value and limitations of radiographs in orthopaedic disease processes. The tarsus and proximal suspensory origin of the metatarsus are commonly imaged regions for lameness. The knowledge gained by comparison between imaging modalities will aid in improving accuracy of radiographic interpretation. OBJECTIVE: To compare the radiographic and MRI findings of the proximal third metatarsal bone (MTIII) and proximal suspensory ligament using MRI as the gold standard. STUDY DESIGN: Retrospective observational study. METHODS: Single hindlimbs of 35 horses with radiographic and high-field (3Tesla (3T)) MRI studies were blindly evaluated by two board-certified veterinary radiologists and a radiology resident. Severity and location of the following parameters were assessed: radiographic MTIII sclerosis and lucent regions, MRI endosteal metatarsal sclerosis and bone marrow lesions, plantar cortical proliferation and resorption, and proximal suspensory desmopathy (PSD). RESULTS: Radiographic osseous changes of MTIII were identified in 54% of limbs, whereas 40% limbs had osseous changes on MRI: 43% of limbs had PSD on MRI. No significant association was found between the presence of radiographic changes in MTIII and PSD on MRI (P = .7). A statistically significant association and positive correlation was found between the severity of radiographic changes and MTIII plantar cortical proliferation and resorption on MRI (P = .01). MAIN LIMITATIONS: Retrospective study with no histopathology. Ability to correlate findings with lameness was limited by the lack of standardised time intervals between onset of lameness, diagnostic analgesia and advanced imaging. CONCLUSION: Radiographic bony changes of proximal MTIII do not reliably predict presence or severity of PSD but are associated with osseous changes at the proximal MTIII. The presence of radiographic changes should warrant additional diagnostics in the region prior to making conclusions about the presence or absence of PSD.

INTRODUCTION/CONTEXTE: La comparaison de la radiographie avec l'imagerie par résonance magnétique (IRM) peut assister dans l'évaluation objective de la valeur et des limites des radiographies pour les processus pathologiques orthopédiques. OBJECTIFS: Comparer les trouvailles radiographiques et d'IRM du métatarsien principal proximal (MTIII) et de la portion proximale du ligament suspenseur du boulet, en utilisant l'IRM comme référence. TYPE D'ÉTUDE: Étude de observationnelle rétrospective. MÉTHODES: Les images radiographiques et d'IRM à haut champ (3 Tesla (3T)) d'un seul membre postérieur chez 35 chevaux ont été évalués à l'aveugle par deux radiologistes vétérinaires diplômés et un résident en radiologie. La sévérité et localisation des paramètres suivants ont été documentés : sclérose et régions lytiques radiographiques du MTIII, sclérose métatarsienne de l'endostéum à l'IRM et lésions de la moelle osseuse, prolifération et résorption plantaire corticale et desmopathie du ligament suspenseur du boulet (DLSB). RÉSULTATS: Des changements radiographiques osseux du MTIII ont été identifiés sur 54 % des membres contre 40% des membres ayant des changements osseux à l'IRM. 43% des membres avaient une DLSB à l'IRM. Aucune association significative n'a pu être identifiée entre la présence de changements radiographiques du MTIII et DLSB à l'IRM (P = 0.7). Une association statistiquement significative et une corrélation positive a été identifiée entre la sévérité des changements radiographiques et la résorption/prolifération plantaires corticales du MTIII à l'IRM (P = 0.01). LIMITES PRINCIPALES: Étude rétrospective sans histopathologie. La possibilité de corréler les données de l'examen de boiterie a été limitée par le manque d'intervalle de temps standardisés entre l'apparition de la boiterie, l'analgésie diagnostique et l'imagerie avancée. CONCLUSIONS: Ces trouvailles démontrent que les changements osseux radiographiques du MTIII proximal ne peuvent prédirent de façon fiable la présence ni la sévérité de DLSB, mais ils sont par contre associés aux changements osseux du MTIII proximal à l'IRM. La présence de changements radiographiques justifie l'utilisation de méthodes diagnostiques additionnelles du MTIII proximal avant de conclure sur la présence ou l'absence de DLSB.

Equine shock wave therapy - where are we now?

Over the past three decades, electrohydraulic extracorporeal shock wave therapy (ESWT) as a treatment modality for equine orthopaedic disorders has sparked exponential interest among practitioners, but its clinical applications are quickly evolving and a current review highlighting modernised equine clinical use is lacking. The objective of this review is to summarise the most current ESWT technology, evidence for its use, proposed mechanisms of action and clinical applications in horses while also highlighting the areas requiring further investigation. The three ways to generate a shock wave are through electrohydraulic, electromagnetic or piezoelectric mechanisms, but over the last decade, electrohydraulic systems have predominated due to the ability to focus and control a therapeutic waveform. Shock waves' primary physical effect is believed to be via mechanotransduction leading to cellular activation and downstream signalling. Experimentally, ESWT's effects on osseous, connective tissue and wound healing via various mechanisms of action have been reported both in the human and veterinary literature. Clinical trials have investigated ESWT's orthopaedic application including osteoarthritis, thoracolumbar pain, navicular syndrome, tendinopathy and proximal suspensory desmopathy, with its concomitant use with biologics representing an area of active research. Direct ESWT protocol comparisons in terms of long-term efficacy with variables of energy, depth and exposed tissue types are still lacking with evidence-based recommendations being largely anecdotal. Technical advancements to facilitate the safe and judicious use of ESWT include human and equine hearing protection, light sedation and/or patient restraint. Efforts to ensure the safe and judicious use of ESWT and its analgesic effects are ongoing.