Ultrasonography of the suspensory ligament branches in yearling and 2-year-old Thoroughbred sales horses: Prevalence, progression of findings and associations with racing performance.

BACKGROUND: Equine suspensory ligament branch (branch) ultrasonography is becoming increasingly commonplace presale. No ultrasonographical branch reference data exists for Thoroughbred sales horses. OBJECTIVES: To define the prevalence of ultrasonographical findings in the forelimb suspensory branches of yearling and 2-year-old sales Thoroughbreds and to analyse associations with racing performance. To track changes in branch 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. Ultrasonography was performed immediately prior to the sales. Ultrasonographical findings relating to branch size, fibrillar pattern, the presence of hyperechoic foci, periligamentar tissue thickness and the adjacent proximal sesamoid bone surface were examined. Associations with racing performance from 2 to 4 years of age were investigated using multivariate regression analyses. Clinical follow-up was sought to ascertain why horses that did not race never started. RESULTS: A total of 593 sales yearlings and 367 2-year-olds had forelimb branch ultrasonography performed. Grade ≥2 fibrillar branch change was present in 8.9% of yearlings and 14.4% of 2-year-olds. A 0.25 cm increase in branch width was associated with a 49-day delayed start to racing careers (P < 0.001, 95% confidence interval (95% CI): 21-77 days). The presence of grade 2 hyperechoic foci was associated with significantly lower total earnings (P = 0.01, 95% CI: $2000-$16 022) and lower earnings per start (P = 0.003, 95% CI: $349-$1718) in United States Dollars. Grade 3 fibrillar branch change had clinically important reductions in the probability of racing, the calibre of racing performance and earnings. Grade 1 fibrillar pattern was associated with significantly higher earnings per start (P = 0.004, 95% CI: $2641-$5759). MAIN LIMITATIONS: The findings are applicable to horses prepared for public auction and deemed fit to be entered for sale. The results may underestimate the proportion of severe lesions in horses not entered for sale. CONCLUSIONS: Reference values specific to young Thoroughbreds have been established. Grade 1 fibrillar branch change should be regarded as an acceptable appearance in sales yearlings and 2-year-olds. Approximately one-third of grade 2 yearling branches progressed to a grade 3 lesion. Evidence of enlarged branch width and grade 2 hyperechoic foci at 2-year-old sales constitute a risk to racing performance.

Concurrent radiological and ultrasonographical findings in the forelimb proximal sesamoid bones and adjacent suspensory ligament branches in yearling and 2-year-old Thoroughbred sales horses.

BACKGROUND: Changes in the proximal sesamoid bones (sesamoids) and the insertional region of the adjacent suspensory ligament branch (branch) are of particular importance in young Thoroughbreds sold at public auction. Little is known about the prevalence of concurrent ultrasonographical branch change, relative to the various grades of radiological sesamoid appearance. OBJECTIVE: To examine the existence of concurrent radiological and ultrasonographical findings in individual sesamoid-branch units in sales horses; to determine whether there are any radiological findings that are consistently accompanied by a particular degree of insertional branch change, and to provide practical recommendations as to when suspensory branch ultrasonography may be warranted in a sales environment. STUDY DESIGN: Prospective cohort study using an enrolled sample. METHODS: Horses were enrolled with consignor permission from a large Thoroughbred yearling sale and five 2-year-old Thoroughbred sales the following year. Data from the radiological evaluation of forelimb sesamoids and the ultrasonographical examination of the adjacent forelimb suspensory branches were described. RESULTS: A total of 2204 yearling forelimb sesamoid-branch units and 1336 2-year-old forelimb sesamoid-branch units were included, from 551 sales yearlings and 334 sales 2-year-olds. The proportion of yearling sesamoids with grade ≤1 vascular channels that had adjacent grade ≥2 fibrillar branch change was 1.2%. The same proportion for 2-year-olds was 3.8%, with medial forelimb sesamoids with grade 1 vascular channels overrepresented in 2-year-olds in this category. In yearlings, 31% of sesamoids with grade 2 vascular channels had adjacent grade ≥2 fibrillar branch change and 59% of sesamoids with grade 3 vascular channels had adjacent grade ≥2 fibrillar branch change. In 2-year-olds, 47% of sesamoids with grade 2 vascular channels had adjacent grade ≥2 fibrillar branch change and 67% of sesamoids with grade 3 vascular channels had the same. Only one yearling sesamoid and one 2-year-old sesamoid with radiological abaxial concavity had grade ≥2 fibrillar branch change. MAIN LIMITATIONS: Hindlimbs were not included. Clinical examinations were not performed and the status of any past or present inflammatory process at the sesamoid-branch enthesis could not be inferred from radiographs and ultrasonographic images alone. CONCLUSIONS: The existence and prevalence of concurrent radiological and ultrasonographical findings in the proximal sesamoid bones and adjacent suspensory ligament branches in yearling and 2-year-old Thoroughbred sales horses has been established. General recommendations have been made for selective branch ultrasonography on the basis of radiological sesamoid appearance. The results support a separate aetiology for radiological sesamoid abaxial concavity that does not primarily involve the suspensory branch insertion.

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.

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.

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.

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.

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.

A polymer network architecture provides superior cushioning and lubrication of soft tissue compared to a linear architecture.

We report the relationships between linear vs. network polymer architecture and biomechanical outcomes including lubrication and cushioning when the polymers are applied to the surface of articulating knee cartilage. Aqueous formulations of the bioinspired polymer poly(2-methacryloyloxylethyl phosphorylcholine) (pMPC) exhibit tuneable rheological properties, with network pMPC exhibiting increased elasticity and viscosity compared to linear pMPC. Application of a polymer network, compared to a linear one, to articulating tissue surfaces reduces friction, lessens tissue strain, minimizes wear, and protects tissue - thereby improving overall tissue performance. Administration of the network pMPC to the middle carpal joint of skeletally mature horses elicits a safe response similar to saline as monitored over a 70 day period.

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.

Gene therapy approaches for equine osteoarthritis.

With an intrinsically low ability for self-repair, articular cartilage injuries often progress to cartilage loss and joint degeneration resulting in osteoarthritis (OA). Osteoarthritis and the associated articular cartilage changes can be debilitating, resulting in lameness and functional disability both in human and equine patients. While articular cartilage damage plays a central role in the pathogenesis of OA, the contribution of other joint tissues to the pathogenesis of OA has increasingly been recognized thus prompting a whole organ approach for therapeutic strategies. Gene therapy methods have generated significant interest in OA therapy in recent years. These utilize viral or non-viral vectors to deliver therapeutic molecules directly into the joint space with the goal of reprogramming the cells' machinery to secrete high levels of the target protein at the site of injection. Several viral vector-based approaches have demonstrated successful gene transfer with persistent therapeutic levels of transgene expression in the equine joint. As an experimental model, horses represent the pathology of human OA more accurately compared to other animal models. The anatomical and biomechanical similarities between equine and human joints also allow for the use of similar imaging and diagnostic methods as used in humans. In addition, horses experience naturally occurring OA and undergo similar therapies as human patients and, therefore, are a clinically relevant patient population. Thus, further studies utilizing this equine model would not only help advance the field of human OA therapy but also benefit the clinical equine patients with naturally occurring joint disease. In this review, we discuss the advancements in gene therapeutic approaches for the treatment of OA with the horse as a relevant patient population as well as an effective and commonly utilized species as a translational model.

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.

Examining the Effects of the Oral Supplement Biota orientalis in the Osteochondral Fragment-Exercise Model of Osteoarthritis in the Horse.

Osteoarthritis (OA) is a common problem in horses. Several oral supplements have been proposed as treatments for horses with OA. The object of this study was to evaluate the use of the oil extract from the seeds of Biota orientalis (BO) for the treatment of experimentally induced OA in horses. OA was induced in 16, 2-5 year old horses in one middle carpal joint on Day 0; the other limb underwent a sham operation. Once daily oral treatment with BO or placebo was initiated on Day 0 and continued to Day 70. All horses were exercised 5 days a week starting on Day 14 through Day 70. The horses were evaluated every other week for lameness and serum GAG concentration as well as weekly synovial fluid assessment. Magnetic resonance imaging was performed on Day 7 and 70. Radiographic changes were assessed on Day 0, 14, and 70. On Day 70 tissue from the middle carpal joint was assessed macroscopically and histologically. All outcome parameters were compared between treatment groups to identify effects of treatment. Compared to placebo a significant decrease was found in synovial fluid prostaglandin E2 concentration and white blood cell counts in horses treated with BO. There was a significant reduction in radiographic scores for subchondral lysis of the radial carpal bone, osteophyte formation, subchondral sclerosis of the radial carpal bone, and total radiographic score for the horses treated with BO. There was no significant difference between treatment groups in clinical lameness findings, MRI findings, macroscopic grading or histologic grading. This study suggests a significant anti-inflammatory effect from oral BO that should be further investigated in clinical OA.

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.

Non-steroidal anti-inflammatory drugs in equine orthopaedics.

Orthopaedic disorders are commonly encountered in equine veterinary medicine, and non-steroidal anti-inflammatory drugs (NSAIDs) play an important role in the management of many equine orthopaedic disorders. There are multiple NSAIDs available for use in horses, including both non-selective and selective NSAIDS, and the body of literature evaluating the efficacy of these medications, their effects on normal and inflamed musculoskeletal tissues, and their side effects is broad. This review aims to summarise the current literature on the use of NSAIDs for equine orthopaedic disorders and examines new and future avenues for the management of inflammation in equine orthopaedics.

Slab fractures of the third carpal bone involving both radial and intermediate facets and outcomes after surgical repair in racing Quarter Horses.

BACKGROUND: Characteristics and outcomes after surgical repair of third carpal bone (C3) slab fractures involving both radial and intermediate facets in racing Quarter Horses are unknown. OBJECTIVES: To describe the pre- and intraoperative characteristics of C3 slab fractures of both radial and intermediate facets in Quarter Horses and to report on the long-term outcomes after internal fixation. STUDY DESIGN: Retrospective case series. METHODS: Case records were collected from racehorses with C3 slab fractures between 2008 and 2020. Inclusion criteria required arthroscopic-guided repair of C3 slab fractures involving both radial and intermediate facets in Quarter Horses. Routine C3 slab fractures (single facet), fractures in other breeds or those repaired with other techniques were excluded. Outcomes were obtained by standardised questionnaire. Data were presented as mean ± SD or as proportions with 95% confidence interval (CI). RESULTS: Of 22 Quarter Horses with C3 slab fractures involving both radial and intermediate facets, 91% (CI 79%-100%; n = 20) were collapsing and 91% (CI 79%-100%; n = 20) had avulsion of the medial palmar intercarpal ligament. Articular cartilage erosion and osteochondral fracture of the radial carpal bone was observed in 91% (CI 79%-100%; n = 20) and 41% (CI 20%-62%; n = 9) cases respectively. At 5.5 ± 3.9 years after surgery, 86% (CI 72%-100%; n = 19) were alive and used for breeding or retirement. Of 18 horses with follow-up >1 year, 39% (CI 16%-61%; n = 7) resumed some athletic activity. MAIN LIMITATIONS: Cases were referred specifically for surgical repair and horses with fractures considered too severe for surgical intervention or euthanasia at owner request were not included. Questionnaire responses are susceptible to recall bias. CONCLUSIONS: Horses with C3 slab fractures of both radial and intermediate facets that are repaired have a favourable prognosis for retirement, breeding and potentially low-level athletic activity.

Corrigendum: Gene therapy approaches for equine osteoarthritis.

[This corrects the article DOI: 10.3389/fvets.2022.962898.].

Immune response to allogeneic equine mesenchymal stromal cells.

BACKGROUND: Mesenchymal stromal cells (MSCs) are believed to be hypoimmunogeneic with potential use for allogeneic administration. METHODS: Bone marrow was harvested from Connemara (n = 1), Standardbred (n = 6), and Thoroughbred (n = 3) horses. MSCs were grouped by their level of expression of major histocompatibility factor II (MHC II). MSCs were then sub-grouped by those MSCs derived from universal blood donor horses. MSCs were isolated and cultured using media containing fetal bovine serum until adequate numbers were acquired. The MSCs were cultured in xenogen-free media for 48 h prior to use and during all assays. Autologous and allogeneic MSCs were then directly co-cultured with responder leukocytes from the Connemara horse in varying concentrations of MSCs to leukocytes (1:1, 1:10, and 1:100). MSCs were also cultured with complement present and heat-inactivated complement to determine whether complement alone would decrease MSC viability. MSCs underwent haplotyping of their equine leukocyte antigen (ELA) to determine whether the MHC factors were matched or mismatched between the donor MSCs and the responder leukocytes. RESULTS: All allogeneic MSCs were found to be ELA mismatched with the responder leukocytes. MHC II-low and universal blood donor MSCs caused no peripheral blood mononuclear cell (PBMC) proliferation, no increase in B cells, and no activation of CD8 lymphocytes. Universal blood donor MSCs stimulated a significant increase in the number of T regulatory cells. Neutrophil interaction with MSCs showed that universal blood donor and MHC II-high allogeneic MSCs at the 6 h time point in co-culture caused greater neutrophil activation than the other co-culture groups. Complement-mediated cytotoxicity did not consistently cause MSC death in cultures with active complement as compared to those with inactivated complement. Gene expression assays revealed that the universal blood donor group and the MHC II-low MSCs were more metabolically active both in the anabolic and catabolic gene categories when cultured with allogeneic lymphocytes as compared to the other co-cultures. These upregulated genes included CD59, FGF-2, HGF, IDO, IL-10, IL-RA, IL-2, SOX2, TGF-ß1, ADAMSTS-4, ADAMSTS-5, CCL2, CXCLB/IL-8, IFNγ, IL-1ß, and TNFα. CONCLUSIONS: MHC II-low MSCs are the most appropriate type of allogeneic MSC to prevent activation of the innate and cell-mediated component of the adaptive immune systems and have increased gene expression as compared to other allogeneic MSCs.

Allogeneic mesenchymal stromal cells for cartilage regeneration: A review of in vitro evaluation, clinical experience, and translational opportunities.

The paracrine signaling, immunogenic properties and possible applications of mesenchymal stromal cells (MSCs) for cartilage tissue engineering and regenerative medicine therapies have been investigated through numerous in vitro, animal model and clinical studies. The emerging knowledge largely supports the concept of MSCs as signaling and modulatory cells, exerting their influence through trophic and immune mediation rather than as a cell replacement therapy. The virtues of allogeneic cells as a ready-to-use product with well-defined characteristics of cell surface marker expression, proliferative ability, and differentiation capacity are well established. With clinical applications in mind, a greater focus on allogeneic cell sources is evident, and this review summarizes the latest published and upcoming clinical trials focused on cartilage regeneration adopting allogeneic and autologous cell sources. Moreover, we review the current understanding of immune modulatory mechanisms and the role of trophic factors in articular chondrocyte-MSC interactions that offer feasible targets for evaluating MSC activity in vivo within the intra-articular environment. Furthermore, bringing labeling and tracking techniques to the clinical setting, while inherently challenging, will be extremely informative as clinicians and researchers seek to bolster the case for the safety and efficacy of allogeneic MSCs. We therefore review multiple promising approaches for cell tracking and labeling, including both chimerism studies and imaging-based techniques, that have been widely explored in vitro and in animal models. Understanding the distribution and persistence of transplanted MSCs is necessary to fully realize their potential in cartilage regeneration techniques and tissue engineering applications.

Colony Forming Potential and Protein Composition of Commercial Umbilical Cord Allograft Products in Comparison With Autologous Orthobiologics.

BACKGROUND: Umbilical cord (UC) connective tissues contain plastic-adherent, colony forming unit-fibroblasts (CFU-Fs) amenable to culture expansion for potential therapeutic use. Recently, UC-derived allograft products have been made available to practitioners in orthopaedics and other specialties, by companies purporting "stem cell"-based healing. However, such marketing claims conflict with existing regulations for these human tissues, generating questions over the cellular and protein composition of current commercially available UC allograft products. PURPOSE: To evaluate commercial UC allograft products for viable cells, CFU-Fs, and protein makeup. STUDY DESIGN: Descriptive laboratory study. METHODS: Five commercial UC allograft products claiming to contain viable, undescribed "stem cells," 2 obtained from UC blood (UCB) and 3 from UC tissue (UCT), were analyzed. Image-based methods were used to measure cell concentration and viability, a traditional CFU-F assay was used to evaluate in vitro behavior indicative of a connective tissue progenitor cell phenotype often referred to as mesenchymal stem/stromal cells, and quantitative immunoassay arrays were used to measure a combination of cytokines and growth factors. Bone marrow concentrate (BMC) and plasma derived from the blood and bone marrow of middle-aged individuals served as comparative controls for cell culture and protein analyses, respectively. RESULTS: Viable cells were identified within all 5 UC allograft products, with those derived from UCB having greater percentages of living cells (40%-59%) than those from UCT (1%-22%). Compared with autologous BMC (>95% viability and >300 million living cells), no CFU-Fs were observed within any UC allograft product (<15 million living cells). Moreover, a substantial number of proteins, particularly those within UCB allograft products, were undetectable or present at lower concentrations compared with blood and bone marrow plasma controls. Interestingly, several important growth factors and cytokines, including basic fibroblast growth factor, hepatocyte growth factor, interleukin-1 receptor antagonist, and osteoprotegerin, were most prevalent in 1 or more UCT allograft products as compared with blood and bone marrow plasma. CONCLUSION: CFU-Fs, often referred to as stem cells, were not found within any of the commercial UC allograft products analyzed, and clinicians should remain wary of marketing claims stating otherwise. CLINICAL RELEVANCE: Any therapeutic benefit of current UC allograft products in orthopaedic medicine is more likely to be attributed to their protein composition (UCT > UCB) or inclusion of cells without colony forming potential (UCB > UCT).

Effects of an articular cartilage lubrication with a viscosupplement in vitro and in vivo following osteochondral fractures in horses.

OBJECTIVE: To assess whether the combination of hyaluronan, sodium chondroitin sulfate, and N-acetyl-d-glucosamine (HCSG) lubricates articular cartilage in vitro and modulates joint lubrication in vivo. ANIMALS: 16 healthy adult horses. PROCEDURES: The effects of HCSG injections on SF lubricant properties and joint health, immediately after injury and 2 weeks later, were analyzed by use an equine osteochondral fracture model of post-traumatic osteoarthritis (OA). Middle carpal joints of adult horses were randomly assigned to 1 of 4 surgical treatment groups as follows: normal nonsurgical group (n = 8), normal sham-surgical group (8), OA-induced surgical group with HCSG injection (8), or OA-induced surgical group with saline (0.9% NaCl) solution injection (8). Synovial fluid was aspirated periodically and analyzed for boundary lubrication function and lubricant molecules. At 17 days, joints were screened for gross pathological changes. RESULTS: Induction of OA led to an impairment of SF lubrication function and diminished hyaluronan concentration in a time-dependent manner following surgery, with HCSG injection lessening these effects. Certain friction coefficients approached those of unaffected normal equine SF. Induction of OA also caused synovial hemorrhage at 17 days, which was lower in joints treated with HCSG. CONCLUSIONS AND CLINICAL RELEVANCE: After induction of OA, equine SF lubricant function was impaired. Hyaluronan-sodium chondroitin sulfate-N-acetyl-d-glucosamine injection restored lubricant properties at certain time points and reduced pathological joint changes.