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An established lipopolysaccharide (LPS) model previously described in Warmbloods, was inconsistent in Standardbred horses, where lameness was not detected despite the presence of synovitis. The present study aimed to determine the dose of LPS from E. coli O55:B5 required to induce mild to moderate lameness following middle carpal joint injection in Standardbred horses and to quantitate the induced lameness over time, with and without anti-inflammatory pre-treatment. In a baseline trial, eight healthy, clinically sound Standardbred horses were used in a rule-based dose-escalation design trial, starting at a dose of 10 endotoxin units (EU). Lameness at trot was evaluated visually and quantitatively (using an inertial-sensor system and pressure plate analysis). Synovial fluid aspirates were analysed for total nucleated cell counts, total protein and prostaglandin E 2 (PGE 2 ). Following 2 months wash-out, the effective LPS-dose determined in the baseline trial was used to evaluate the effect of anti-inflammatory treatment. A mixed model for repeated measures with horse as random effect was used for analysis. After injection of 10 EU LPS, the desired degree of lameness was observed in the baseline trial, with maximal lameness at post-injection hour (PIH) 4, followed by a rapid decline and return to baseline by PIH 48. No lameness was observed following pre-treatment with meloxicam. In synovial fluid, PGE 2 was significantly higher at PIH 8 and PIH 24 in the baseline trial compared with following meloxicam pre-treatment. In conclusion, injection of the middle carpal joint with 10 EU LPS consistently induces a transient lameness and synovitis in Standardbred horses., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

At Fédération Equestre Internationale (FEI) competitions, horses undergo veterinary inspection for judgement of 'fit-to-compete'. However, FEI Veterinary Delegates (VDs) often differ in opinion. The aim of the present study was to evaluate intra- and inter-observer agreements of fit-to-compete judgement and compare these with objective gait analysis measurements. Twelve horses were evaluated by three experienced VDs and one veterinary specialist and video-recorded for re-evaluation later. Simultaneously, quantitative gait analysis measurements were acquired. Inter-observer agreement during live evaluations was fair (κ = 0.395, 58% agreement). Intra-observer agreement between live observations and videos at one and 12 months was 71% and 73% respectively. Sensitivity and specificity of motion symmetry measured with quantitative gait analysis system were 83.3% and 66.7% respectively, against the consensus of all observers as a reference. These findings might suggest that more VDs should be used to adequately judge fit-to-compete. Quantitative-gait-analysis may be useful to support decision making during fit-to-compete judgement., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Cartilage repair remains a major challenge and treatment of (osteo)chondral defects generally results in poor quality fibrous repair tissue. Our approach aims to address some of the major biomechanical issues encountered in scaffold-based cartilage repair, such as insufficient stiffness of the scaffolds, step formation at the interface with the native tissue and inadequate integration with the original tissue. Two osteochondral defects were created on the medial femoral trochlear ridge in each stifle of six Shetland ponies. The defects were filled with a bi-layered implant consisting of a polyetherketoneketone (PEKK) bone anchor and a polyurethane elastomer. The defects in the contralateral joint served as unfilled controls. After 12 weeks, the ponies were euthanased and tissues were evaluated macroscopically and using micro-computed tomography, histology and immunohistochemistry. Post-operative recovery was good in all ponies and minimal lameness was observed. After 12 weeks, the proximally located plug was partially covered (mean±standard deviation [SD] percentage surface area covered 72.5±19.7%) and the distal plug was nearly completely covered (mean±SD percentage surface area covered 98.5±6.1%) with stiff and smooth repair tissue. Histology and immunohistochemistry confirmed that the repair tissue was well connected to the native cartilage but contained negligible amounts of collagen type II and glycosaminoglycans (GAGs). The repair tissue was stiff and fibrous in nature and presented a nearly flush surface with the surrounding native cartilage distally. This approach therefore resolves a number of issues related to scaffold-based cartilage repair and compares favourably with results of several other studies in large animal models. However, long-term follow-up is needed to evaluate the true potential of this type of implant., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Extracellular vesicle (EV) concentration, characteristics and function in equine synovial fluid (SF) during normal growth and development has not previously been studied. Isolation of EVs was performed in SF from three healthy foals and two adult horses by differential ultracentrifugation (10,000g and 200,000g); EVs were purified by sucrose density gradient floatation and analysed by high-resolution flow cytometry (FCM), buoyant density and western blotting. Additionally, repeated biomarker analysis of sulphated glycosaminoglycans (GAG), matrix metalloproteinase (MMP), C-terminal crosslinked telopeptide type II collagen (CTX-II), collagenase cleaved neopeptide type II collagen (C2C) was performed in SF from 10 foals and six adult horses. In contrast with the quantitative EV profile, the biomarker profile in SF from juvenile joints was substantially different from that in SF from adult animals. However, there were qualitative differences in the high-resolution FCM scatter plots. Future in-depth functional analyses may reveal differences between juvenile and mature EVs in SF., (Copyright © 2018. Published by Elsevier Ltd.)

Quantitative gait analysis has the potential to offer objective and unbiased gait information that can assist clinical decision-making. In recent years, a growing number of gait analysis systems have come onto the market, highlighting the demand for such technology in equine orthopaedics. However, it is imperative that the measured variables which are used as outcome parameters are supported by scientific evidence and that the interpretation of such measurements is backed by a proper understanding of the biomechanical principles of equine locomotion. This review, which is based on studies on experimentally induced lameness, summarises the currently most widely used methods for gait analysis and the available evidence concerning gait parameters that can be used to quantify gait changes due to lameness. These are discussed regarding their current and future potential for routine clinical application., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Although phenylbutazone (PBZ) is commonly used in equine orthopaedic practice, little is known about its in vivo effects on joint inflammation and cartilage turnover. This study investigates the effects of PBZ on inflammatory parameters, matrix metalloproteinase (MMP) activity and cartilage biomarkers in equine joints with acute synovitis. In a two-period cross-over study, transient synovitis was induced at T = 0 h in the middle carpal joint of seven ponies by lipopolysaccharide (LPS) injection. Ponies received PBZ (2 mg/kg PO twice daily) or placebo for 1 week, starting at T = 2 h. Arthroscopic assessment of the middle carpal joint was performed at T = -504, 48 and 672 h. Synovial fluid (SF) was sampled at T = -504, 0, 8, 24, 48, 168, 336 and 672 h and analysed for leukocytes and total protein, substance P, general MMP activity, glycosaminoglycans (GAG), collagen II cleavage marker C2C and synthesis marker CPII. Markers in PBZ- vs. placebo-treated joints were compared over time using a linear mixed model. LPS injection caused marked transient synovitis without visible cartilage changes. Substance P and general MMP activity were not significantly reduced by PBZ treatment, nor were SF GAG or C2C concentrations at any time point. Concentration of CPII was significantly lower at T = 24 and 168 h in PBZ treated joints compared to placebo. Although PBZ is clinically effective in treating acute synovitis, it does not limit inflammation-induced cartilage catabolism and may transiently reduce collagen anabolism as evidenced by SF markers., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Arthroscopy enables direct inspection of the articular surface, but provides no information on deeper cartilage layers. Optical coherence tomography (OCT), based on measurement of reflection and backscattering of light, is a diagnostic technique used in cardiovascular surgery and ophthalmology. It provides cross-sectional images at resolutions comparable to that of low-power microscopy. The aim of this study was to determine if OCT is feasible for advanced clinical assessment of lesions in equine articular cartilage during diagnostic arthroscopy. Diagnostic arthroscopy of 36 metacarpophalangeal joints was carried out ex vivo. Of these, 18 joints with varying degrees of cartilage damage were selected, wherein OCT arthroscopy was conducted using an OCT catheter (diameter 0.9 mm) inserted through standard instrument portals. Five sites of interest, occasionally supplemented with other locations where defects were encountered, were arthroscopically graded according to the International Cartilage Repair Society (ICRS) classification system. The same sites were evaluated qualitatively (ICRS classification and morphological description of the lesions) and quantitatively (measurement of cartilage thickness) on OCT images. OCT provided high resolution images of cartilage enabling determination of cartilage thickness. Comparing ICRS grades determined by both arthroscopy and OCT revealed poor agreement. Furthermore, OCT visualised a spectrum of lesions, including cavitation, fibrillation, superficial and deep clefts, erosion, ulceration and fragmentation. In addition, with OCT the arthroscopically inaccessible area between the dorsal MC3 and P1 was reachable in some cases. Arthroscopically-guided OCT provided more detailed and quantitative information on the morphology of articular cartilage lesions than conventional arthroscopy. OCT could therefore improve the diagnostic value of arthroscopy in equine orthopaedic surgery., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

This manuscript describes a new classification of the various joint-related lesions that can be seen in the young, growing horse based on their anatomical and functional aetiopathogenesis. Juvenile osteochondral conditions (JOCC) is a term that brings together specific disorders according to their location in the joint and their biomechanical origin. When a biomechanical insult affects the process of endochondral ossification different types of osteochondrosis (OC) lesions may occur, including osteochondral fragmentation of the articular surface or of the periarticular margins, or the formation of juvenile subchondral bone cysts. In severe cases, osteochondral collapse of the articular surface or the epiphysis or even an entire small bone may occur. Tension on ligament attachments may cause avulsion fractures of epiphyseal (or metaphyseal) ossifying bone, which are classified as JOCC, but do not result from a disturbance of the process of endochondral ossification and are not therefore classified as a form of OC. The same applies to 'physitis' which can result from damage to the physeal growth plate., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Intra-articular injection of opioids provides analgesia in painful equine joints and μ-opioid receptors (MORs) have been demonstrated in equine synovial membranes. The aim of this study was to determine whether acute inflammatory conditions will lead to up-regulation of MOR in equine synovial membranes and whether anti-inflammatory treatment can prevent any such upregulation. In a two-period, blinded, placebo-controlled randomised cross-over design, lipopolysaccharide (LPS, 1.0 ng) was injected into the left or right middle carpal joint of seven healthy ponies. Arthroscopy and synovial membrane biopsy was performed under general anaesthesia at baseline, 48 h (T48) and 672 h (T672) after LPS injection, with ponies assigned to receive either phenylbutazone (PBZ 2.2mg/kg PO BID) or placebo from 2h post-LPS. Ponies were scored for pain and lameness. Repeated synovial fluid samples were obtained and the degree of synovitis scored both macroscopically and microscopically. The density and staining pattern of MOR-like protein in synovial membrane biopsies over the course of the synovitis with or without PBZ treatment was evaluated using immunohistochemical techniques. LPS injection consistently induced a severe transient synovitis. Pain and lameness were significantly attenuated by treatment with PBZ. Up-regulation of MOR-like protein in the inflamed equine synovial membrane could be demonstrated in the placebo treated animals, but not in the PBZ-treated animals overall, although there were no significant differences at any individual time-point between the two groups. It was concluded that acute inflammation will up-regulate MOR, while anti-inflammatory treatment will attenuate this response., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Although alterations in biomarkers of cartilage turnover in synovial fluid (SF) have been demonstrated in horses with osteochondrosis (OC), there have been few investigations of such alterations in animals <1 year old. In this study tarsocrural SF samples from foals aged 18, 22 and 52 weeks of age were assessed for: (1) 'turnover' biomarkers of type II collagen (CPII and C2C) and proteoglycan (CS846 and glycosaminoglycans [GAG]); (2) matrix metalloproteinase (MMP) activity; (3) insulin-like growth factor (IGF)-1; (4) transforming growth factor (TGF)-β1; (5) prostaglandin (PG) E(2); and (6) leukotriene B(4). Using a linear mixed model, the concentration of biomarkers was compared between animals that developed or did not develop radiographic evidence of OC at 24 or 48 weeks of age. The CPII:C2C ratio tended to be higher in OC-affected joints compared to controls at all ages, and this difference was statistically significant at 22 weeks of age. The concentrations of CS846 and IGF-1, and the CS846:GAG ratio were reduced in OC-affected joints relative to controls at 18 weeks of age only. At 52 weeks of age, the PGE(2) concentration was lower in joints with OC. Overall, there appears to be a consistent anabolic shift in type II collagen turnover in juvenile joints affected by OC. Aberrant proteoglycan turnover is not a hallmark of the late repair of this lesion but reduced concentrations of IGF-1 in SF may be associated with early-stage lesions., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Saddle-fit is recognised as an important factor in the pathogenesis of back problems in horses and is empirically being evaluated by pressure measurements in clinical practice, although not much is known about the validity, reliability and usability of these devices in the equine field. This study was conducted to assess critically a pressure measurement system marketed for evaluating saddle fit. Validity was tested by calculating the correlation coefficient between total measured pressure and the weight of 28 different riders. Reliability and discriminative power with respect to different saddle fitting methods were evaluated in a highly standardised, paired measurement set-up in which saddle-fit was quantified by air-pressure values inside the panels of the saddle. Total pressures under the saddle correlated well with riders' weight. A large increase in over-day sensor variation was found. Within trial intra-class correlation coefficients (ICCs) were excellent, but the between trial ICCs varied from poor to excellent and the variation in total pressure was high. In saddles in which the fit was adjusted to individual asymmetries of the horse, the pressure measurement device was able to detect correctly air-pressure differences between the two panels in the back area of the saddle, but not in the front area. The device yielded valid results, but was only reliable in highly standardised conditions. The results question the indiscriminate use of current saddle pressure measurement devices for the quantitative assessment of saddle-fit under practical conditions and suggest that further technical improvement may be necessary.

Subchondral bone provides structural support to the overlying articular cartilage and plays an important role in osteochondral diseases. There is growing insight that the mechanical features of bone are related to the biochemistry of the collagen network. In this study the effect of exercise on water, calcium and the collagen network (total collagen, lysyl-hydroxylation, hydroxylysylpyridinoline, and lysylpyridinoline crosslinks) of subchondral bone at two differently loaded sites (site 1: intermittently loaded; site 2: constantly loaded) is investigated in foals. Exercise influenced calcium content and levels of both types of crosslinks at site 1, but had no influence on site 2. There was no concomitant increase in lysyl-hydroxylation level with the rise in crosslinks. Levels of lysyl-hydroxylation and lysylpyridinoline crosslinking were lower at site 1 than at site 2. It is concluded that exercise affects the post-translational modifications of the collagen component of subchondral bone. Loading also appears to play a role in site-related topographical differences. The lack of any relation between the sum of pyridinoline crosslinks and the amount of triple helical hydroxylysine gives support to a recent hypothesis that lysyl-hydroxylation of the triple helix and the telopeptides are under separate control., (Copyright 2000 Harcourt Publishers Ltd.)