Search

P>Reasons for performing study:Damage to the flexor tendons, particularly the superficial digital flexor tendon (SDFT), is one of the most common musculoskeletal injuries sustained by horses competing in all disciplines. Our previous work has shown that SDFTs from different individuals show a wide variation in mechanical strengths; this is important clinically as it may relate to predisposition to injury. The high mechanical strength of tendon relies on the correct orientation of collagen molecules within fibrils and stabilisation by the formation of chemical cross-links between collagen molecules. It is not known whether the variation in SDFT mechanical properties between individuals relates to differences in collagen cross-link levels.Hypothesis:Enzyme-derived, intermolecular cross-linking of tendon collagen correlates with mechanical properties of the SDFT.Methods:SDFTs were collected from 38 horses and mechanically tested to failure. Structural and material properties were calculated from the load/deformation plot and cross-sectional area for each tendon. Following mechanical testing, pyrrolic cross-link levels were measured using a spectrophotometric assay for Ehrlich's reactivity and pyridinoline levels were quantified by HPLC. Cross-link levels were correlated with mechanical properties and statistical significance tested using a Pearson's correlation test.Results:Pyrrole cross-link levels showed a significant positive correlation with ultimate stress (P = 0.004), yield stress (P = 0.003) and elastic modulus (P = 0.018) of the tendons, despite being a minor cross-link in these tendons. There was no significant correlation of mechanical properties with either hydroxylysyl- or lysyl-pyridinoline levels.Conclusions:Given the low absolute levels of pyrrole, we suggest that the correlation with high mechanical strength is through an indirect mechanism. Understanding the nature of the relationships between pyrrole cross-links, other matrix characteristics and tendon material properties may allow development of strategies to identify horses at risk from tendon injury and be of value in informing training practices.

P>Reasons for performing the study:Highly prevalent superficial digital flexor tendon (SDFT) injury results in compromised tendon function through fibrosis and high frequency of re-injury due to altered biomechanical function. This study investigated the consequences of SDF tendinopathy on limb mechanics in relation to the mechanical properties of injured tendon.Objectives:To develop and validate a noninvasive in vivo assessment of tendon mechanics to investigate the effect of recent SDFT injury on limb stiffness index, providing an objective method to assess quality of healing.Hypotheses:Limb stiffness index would reduce as a consequence of SDFT injury and progressively increase during tendon healing and correlate with in vitro mechanical properties of the respective SDFTs.Methods:Kinematic analysis was performed at walk in 10 horses that had sustained career-ending SDFT injury. Stiffness index was derived from limb force recorded via a series of force plates and measurement of change in metacarpophalangeal joint angle using 3D motion analysis software. Horses were subjected to euthanasia 7 months after injury, the SDFTs removed and subjected to nondestructive in vitro mechanical testing.Results:Limb stiffness index was reduced following SDFT injury in comparison with the contralateral limb and increased during the convalescent period, approximating that of the contralateral limb by 7 months post injury. There was a significant positive correlation between in vivo limb stiffness index and in vitro SDFT stiffness.Clinical relevance:The ability to assess and monitor SDFT mechanical competence through limb stiffness measurement techniques in horses recovering from SDFT injury and the possibility of corroborating this with functional tendon healing may permit a more objective and accurate assessment of optimal tendon repair in the horse. This technique may be a useful method for assessing the efficacy of treatment regimens for tendinopathy and could be utilised to predict time to safe return to performance or re-injury.

P>Reasons for performing study:Mesenchymal stem cell (MSC) therapy for orthopaedic disease is being used with increasing frequency; there is a need to define a safe, reliable and effective technique for the recovery of MSCs from the sternum of the horse.Objectives:To describe an optimised safe technique for obtaining bone marrow-derived MSCs from the sternum of the Thoroughbred horse.Methods:The anatomical relationship of the sternum with the heart and internal anatomy was demonstrated in cadavers. Sternal anatomy was evaluated ultrasonographically and after midline sectioning. Sternebrae were examined histologically after aspiration to determine the effect of needle insertion. The quality of the aspirate was evaluated as the number of colony-forming units from sequential and separately aspirated 5 ml aliquots and assessed for their multipotency using trilineage differentiation.Results:The optimal safe location for the needle was the 5th sternebra because it had a safe dorsoventral thickness and was cranial to the apex of the heart. This sternebra could be reliably identified ultrasonographically. Aspirates could also be obtained from the 4th and 6th sternebrae, although the former is between the front limbs and the latter closer to the heart. Minimal disruption of the internal bony architecture was seen after needle insertion through the thin outer cortex and the first 5 ml aliquot contained the greatest number of colony-forming units of mesenchymal stem cells with trilineage capabilities.Conclusions:Accurate placement of a Jamshidi needle into the medullary cavity of the 4th-6th individual sternebrae is facilitated by the use of ultrasonography and enables aspiration of bone marrow reliably with minimal damage to the sternum and risk to the horse.Potential clinical relevance:Sternal marrow aspiration as described is a safe and reliable technique to obtain MSCs for orthopaedic cell-based therapies.

In human athletes, conditioning, training and competition are commenced before skeletal maturity. Yet in equine athletics, racing of young (age 2 years) horses remains contentious. Tendon injury persists as major causes of wastage in equine athletes. Minimising injury and associated welfare issues could involve a radical approach to the timing and implementation of conditioning and training. Tendons were examined from Thoroughbreds, Dutch Warmblood foals, working horses and also a group of wild horses to evaluate effects of age, function and exercise. Gross mechanical properties did not differ significantly with age or exercise, but showed a high variance within each group. Mechanical properties of tendon tissue showed significant differences as a function of age and location. The collagen fibril crimp angle and length showed a regional reduction in the central core with exercise and age, with a synergistic effect. Regional differences in collagen fibril diameter were seen in long-term exercised older horses, but not in short-term exercised, or younger, horses. The higher proportion of small fibrils in the central region of the long-term exercised horses did not correlate with new collagen formation and therefore appear to result from disassembly of the larger diameter fibrils. Fibril diameter distributions were influenced by exercise regimens in the growing foal. Changes in molecular composition occurred in longer-term exercise and older horses, in the centre of the tendon, with higher levels of type III collagen and changes in glycosaminoglycan (GAG) content. Cartilage Oligomeric Matrix Protein (COMP) levels also appear to be modulated by age, function and superimposition of exercise. These changes were all exacerbated with age and exercise, suggesting appropriate exercise in young horses may lead to a lower incidence of injury than in older horses. An hypothesis is advanced that immature tendon can respond to exercise while mature tendon has limited, if any, ability to do so. These findings support potentially controversial earlier conditioning and racing of younger, rather than older, equine athletes.

Most skeletal tissues are thought to adapt to the mechanical environment they experience. While this has been demonstrated for muscle and bone, previous studies in the mature horse have failed to demonstrate adaptation in the superficial digital flexor tendon (SDFT), which suffers a high frequency of injury. This study tested the hypothesis that imposed exercise during growth would result in an increase in SDFT cross-sectional area (CSA). Fourteen Thoroughbred foals were divided into 2 sex-matched groups. A control group received 4 h pasture exercise and an exercise group had the same amount of pasture exercise with an additional short period of treadmill exercise daily from age 2-15 months. Activity at pasture was assessed objectively using a visual system. There was no significant difference in pasture activity between groups, although males were more active than females. The exercise programme resulted in a significantly larger tendon CSA in the exercise group at several, but not all, timepoints, which may be attributed to levels of variance. However, there was a significantly greater rate of increase in tendon CSA as a function of time in the exercised compared to the control group. This is the first evidence to suggest that tendon development can be modulated by exercise during growth in the horse, potentially increasing the ability of tendon to withstand the rigours of later athletic activity.

Reasons for performing study: Osteoarthritis (OA) is a common cause of distal tarsal pain, but disease development is poorly understood. Awareness of normal tarsal structure and function is important in order to understand the pathogenesis of OA. Thickening of the subchondral bone (SCB) plate has been related to the development of OA, but SCB plate patterns in the equine tarsus have not been documented.Hypotheses: There is a repeatable pattern of SCB thickness across the distal tarsal joints, and specifically that thickness would be greatest dorsally and laterally.Methods: Twenty cadaver tarsi were collected from mature horses that had undertaken low-level exercise only with no history of hindlimb lameness. Magnetic resonance images were acquired using a high-resolution sagittal 3-dimensional T1-weighted spoiled gradient echo sequence. Subchondral bone thickness was measured on sagittal images at dorsal and plantar locations on the proximal and distal aspects of the central (CT) and third (T3) tarsal bones and proximal aspect of the third metatarsal bone (MO).Results: On the proximal aspect of CT, medial and lateral SCB thickness were significantly greater than midline. On the distal aspect of CT and T3 and proximal Mt3, lateral SCB thickness was significantly greater than medial and midline sites. Dorsal SCB thickness was greatest on the proximal and distal aspects of CT and proximal Mt3. Subehondral bone accounted for a greater proportion of CT and T3 on the dorsal aspect than the plantar.Conclusions: There is a repeatable pattern of SCB thickness in the distal tarsal bones of horses with no history of hindlimb lameness. This reflects the pattern of loading across the joints.Potential relevance: This study provides evidence of a consistent osteochondral pattern in the equine tarsus for reference in identification of osteoarticular pathologies.

Conditioning by early training may influence the composition of certain musculoskeletal tissues, but very few data exist on its effect during growth on tendon structure and function.To investigate whether conditioning exercise in young foals would lead to any ultrasonographically detectable damage to the superficial digital flexor tendon or an increase in cross-sectional area (CSA).Thirty-three Thoroughbred foals reared at pasture were allocated to 2 groups: control (PASTEX) allowed exercise freely at pasture; and CONDEX, also at pasture, began conditioning exercise from mean age 21 days over 1030 m on a purpose-built oval grass track, for 5 days/week until mean age 18 months. Foals were observed daily, and underwent orthopaedic examination monthly. Ultrasonographic images of the superficial digital flexor tendon (SDFT) at the mid-metacarpal level of both forelimbs were obtained in all foals at ages 5, 8, 12, 15 and 18 months. CSA was validated (r(2) = 0.89) by determining CSA from digital photographs of the transected SDFT surface from 12 of the horses necropsied at age 17.1 months.here was no clinical or ultrasonographic evidence of tendonopathy in either group and the greatest increase in mean CSA in both groups occurred between age 5 and 8 months. Across all age categories, there was no significant difference in mean CSA between the left and right limbs, or colts and fillies; there was a trend towards a larger CSA in the CONDEX group (P = 0.058).There was no conclusive evidence for a structural adaptive hypertrophy of the SDFT, probably because the regimen was insufficiently rigorous or because spontaneous pasture exercise may induce maximal development of energy storing tendons.A moderate amount of early conditioning exercise against a background of constant exercise at pasture is not harmful to the development of the flexor tendons.

Bone mineral density (BMD) was measured by dual x-ray absorptiometry (DXA) in the carpus of 6 Thoroughbreds age 24 months that were exercised on the treadmill for 4.5 months, and of 6 matched control horses. A sagittal slab of known thickness was sawn from one or more of radius, radial carpal (Cr), third carpal (C3), and third metacarpal (Mc3) bones. After the 4.5 month treadmill exercise, there was a significantly greater volumetric BMD in the dorsal aspect of C3 (P = 0.006). Treadmill-exercised horses also had higher BMD in the dorsodistal aspect of the radius (P = 0.03), the dorsal half of Mc3 (P = 0.012), and the dorsal aspect of Cr (P = 0.04). In other areas of interest (AOI) studied on the palmar aspect of the bones, significant increases in exercised horses did not occur. These sizeable changes in bone mass occurred within a few months of initiation of exercise. With technical development, DXA could be applied in vivo for investigation of changes in bone mass of horses which are growing and exercising under various regimes. The data are also relevant to the development of scientifically based training protocols.

Processes involved in equine carpal osteochondral injury have not been established. In other species, fibronectin appears important in chondrocyte-matrix interactions, and levels are increased in osteoarthritis. This investigation aimed to (a) describe fibronectin immunoreactivity in the middle carpal joint of 2-year-old Thoroughbreds, (b) determine topographical variations, (c) compare strenuously trained (Group 1) or gently exercised horses (Group 2) and (d) describe sites with early osteoarthritis. Group 1 (n = 6) underwent a 19 week high intensity treadmill training programme. Group 2 (n = 6) underwent 40 min walking until euthanasia. Dorsal and palmar sites on radial, intermediate and third carpal articular surfaces were prepared. Immunohistochemistry was performed using a biotin-streptavidin/peroxidase method. Cross-reactivity of rabbit antihuman fibronectin antiserum with equine fibronectin was confirmed using Western blotting. Results showed: (a) fibronectin was present primarily in pericellular and interterritorial matrix locations, (b) dorsal sites had zonal immunoreactivity compared to palmar sites, (c) Group 1 dorsal radial carpal cartilage had increased superficial staining compared to Group 2 and (d) fibrillated cartilage showed increased intracellular and local matrical immunoreactivity (superficial zone). These findings suggest topographical and exercise-related variations in fibronectin distribution, and indicate equine fibronectin is localised at sites of cartilage degeneration and released into the matrix by chondrocytes in the local area.

This study was undertaken to test the hypothesis that collagen fibrils, the submicroscopic units of strength in tendon, would hypertrophy in response to a specific defined training programme. Fibril diameters were measured in central and peripheral regions of superficial digital flexor tendon (SDFT) samples from five 18-month-old horses which underwent a subsequent 18 month training programme and 6 age- and sex-matched controls. Central region fibrils from the trained horses had a mass-average diameter (MAD) of 105.3 nm, which was significantly lower (P < 0.01) than that of 131.7 nm for the same region in the control horses. This reduction in fibril diameter in the region of tendon which is predisposed to injury was interpreted as evidence of microtrauma, as it implies the region is weakened by the training regimen. Repeated episodes of microtrauma may accumulate and eventually result in degenerative lesions and clinical tendonitis.

Biochemical markers of bone metabolism were analysed in serum samples obtained from 60 horses with no history of orthopaedic disease (age 3 months-20 years). Serum levels of the carboxyterminal propeptide of type I procollagen (PICP), a marker of bone formation and the pyridinoline cross linked telopeptide domain of type I collagen (ICTP), a putative marker of bone resorption, were measured by radioimmunoassay (RIA). Serum levels of the bone specific isoenzyme of alkaline phosphatase (BALP), another marker of bone formation, were measured by a wheatgerm agglutinin affinity (WGA) method. Total alkaline phosphatase levels were also determined. Serum levels of PICP were significantly correlated with bone ALP (r = 0.78, P < 0.0001) and ICTP (r = 0.87, P < 0.0001). ICTP levels also correlated significantly with bone ALP (r = 0.81, P < 0.0001). However, total alkaline phosphatase did not correlate significantly with PICP, ICTP and BALP in horses over 1 year of age. There was an inverse correlation between serum levels of all biochemical markers and age of animals, with the most significant changes seen over the first 2 years. In animals less than 1 year of age, the reference ranges (mean +/- s.d. 1.96) were as follows: PICP 1216-2666 micrograms/l, ICTP 13.8-26.7 micrograms/l, bone ALP 134-288 u/l and total ALP 223-498 u/l. In 2-year-olds, the equivalent reference ranges were: PICP 550-1472 micrograms/l, ICTP 7.96-22.8 micrograms/l, bone ALP 32.7-125 u/l and total ALP 134-238 u/l.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of osteochondral defect location and post-operative walking exercise on structural repair and recovery of joint function were examined in the midcarpal joints of ponies. Functional recovery was monitored by measuring ground reaction forces using a force plate. Structural repair was evaluated histologically and by measuring the total collagen and uronic acid content and relative proportions of Type I and II collagen in the repair tissue. Central defects tended to cause a more marked functional disturbance but were repaired with fibrocartilage rather than fibrous tissue in 3 out of 6 ponies while marginal defects were repaired almost exclusively with fibrous tissue. There was no significant difference between defect locations with regard to the biochemical measures evaluated. Exercise produced no beneficial effect on structural repair of the defects. Secondary 'kissing' lesions in the third carpal bones opposite the defects appeared grossly more severe in the exercised ponies which also showed a trend to more marked functional disturbance.

REASON FOR PERFORMING STUDY The injury-prone, energy-storing equine superficial digital flexor tendon (SDFT) of the mature performance horse has a limited ability to respond to exercise in contrast with the noninjury-prone, anatomically opposing common digital extensor tendon (CDET). Previous studies have indicated low levels of cellular activity in the mature SDFT, but in foal tendons the tenocytes may still have the ability to adapt positively to increased exercise. OBJECTIVES To measure tenocyte densities and types in histological sections from the SDFT and CDET of horses from controlled long-term, short-term and foal exercise studies. METHODS Specimens were collected from mid-metacarpal segments of the CDET and SDFT for each horse and processed for histology; central and peripheral regions of the SDFT cross-section were analysed separately (SDFTc, SDFTp). Tenocyte nuclei were counted in a total area of 1.59 mm(2) for each tendon region in each horse. Each nucleus was classified as type 1 (elongate and thin), type 2 (ovoid and plump) or type 3 (chondrocyte-like); type 1 cells are proposed to be less synthetically active than type 2 cells. RESULTS No significant differences were noted between exercise and control groups in any of the studies, with the exception of an exercise-related reduction in the proportion of type 1 tenocytes for all tendons combined in the long-term study. There were tendon- and site-specific differences in tenocyte densities and proportions of type 1 and 2 cells in all 3 studies. CONCLUSIONS AND POTENTIAL RELEVANCE There was no indication that exercise increased tenocyte density or proportions of the (theoretically) more active type 2 cells in immature horses (short-term and foal studies), perhaps because the training regimens did not achieve certain threshold strain levels. In the foal study these findings can still be interpreted positively as evidence that the training regimen did not induce subclinical damage.

No data exist on the intensity of exercise required or on possible harmful effects of increasing exercise in foals over the natural level when free at pasture.To investigate whether an increase in workload over free pasture exercise in the period from directly after birth to the start of training is tolerated by Thoroughbred (TB) foals without increasing injury rate or producing other undesired side effects.Thirty-three TB foals were allocated to one of 2 exercise groups directly after birth. One group (PASTEX) was raised on pasture and the other (CONDEX) kept under identical circumstances, but was additionally subjected to an exercise protocol of gradually increasing intensity. Foals were monitored periodically and scored for the presence of clinical signs related to the musculoskeletal system (joint effusion, pain at flexion, occurrence of physeal swelling), and radiographs taken at the end of the conditioning phase. Also, behavioural studies were performed to detect any changes in behaviour related to the exercise programme. Cortisol levels were measured in both groups, to assess the level of stress.Workload in the CONDEX group was significantly higher than in the PASTEX group (approximately 30%). Conditioning increased the likelihood for joint effusion in the antebrachiocarpal joint, but reduced tarsocrural effusion and physeal swelling at the lateral distal radius, the third metacarpal bone (medial aspect) and lateral and medial aspects of the third metatarsal bone.The 30% increase in workload did not affect the animals' welfare, effects of conditioning exercise on clinical musculoskeletal health were few and there were no adverse effects.This study supports the feasibility of imposing early conditioning exercise in horses and is a benchmark for its effects on the development of equine musculoskeletal tissues.

Summary Reasons for performing study: Understanding of the development of pathology and source of pain in distal tarsal osteoarthritis is poorly understood. Magnetic resonance imaging is often used in the analysis of human osteoarthritis (OA) because it is sensitive to early changes. Hypothesis: In association with distal tarsal joint (DTJ) pain, there will be an alteration in the characteristic subchondral bone (SCB) thickness pattern of horses with no history of pain when subjected to low-level exercise. Methods: Sixteen cadaver tarsal joints were collected from 9 mature horses with a history of tarsal pain and radiographic evidence of OA; 3 cadaver tarsi were collected from 2 mature horses with a history of tarsal pain and no radiographic abnormality. Magnetic resonance images were acquired using high-resolution sagittal 3D T1 weighted spoiled gradient echo sequence. Subchondral bone thickness was measured on sagittal images at dorsal and plantar locations on the proximal and distal aspects of the central (CT) and third (T3) tarsal bones and proximal aspect of the third metatarsal bone (MT3). Results: In tarsi with radiographic evidence of OA medial and lateral SCB thicknesses were greater than midline on the proximal and distal aspects of CT and T3. Lateral SCB thickness was greater than medial on the proximal aspect of MT3. There was an increase in SCB thickness at the majority of sites compared with normal horses. There were too few joints in the group without radiographic changes to analyse statistically. In painful tarsi SCB thickness was greater medially than laterally at all sites. In horses without tarsal pain all lateral sites had greater SCB thickness, except the proximal aspect of CT. Conclusions: There is alteration of normal SCB thickness patterns in painful tarsi. Different thickness patterns could represent different types of pathological processes. Potential clinical relevance: Further work is required to elucidate the pathological processes leading to OA of the DTJs.

Reasons for performing study: Overstrain injuries to the superficial digital flexor tendon (SDFT) and suspensory ligament (SL) are among the most common musculoskeletal injuries which contribute to the considerable wastage of racing Thoroughbreds. Many epidemiological studies have demonstrated the prevalence of and risk factors for tendon injury when racing but have not included those injuries sustained during training. However, since tendon injury during training is seen commonly in clinical practice, it is appropriate to determine the overall prevalence of tendon injury sustained during both training and racing. Objective: To determine the prevalence of overstrain injury to the SDFT and SL during training and racing among Thoroughbred flat racehorses in Japan in 1999. Methods: A retrospective study was performed using a sample population of 10,262 Thoroughbred racehorses. The medical information database of Thoroughbred racehorses registered by the Japan Racing Association (JRA) in 1999 was analysed for SDFT and SL overstrain injury diagnosed by a veterinarian employed by JRA during training and racing. Jump racehorses were excluded from this study. Results: The prevalence of forelimb SDFT tendonitis and SL desmitis was 11.1% (1130 cases) and 3.61% (370 cases) of the population, respectively. In the hindlimb, there were 0.06% (6 cases) and 0.14% (14 cases), respectively. Risks of SDF tendonitis in the forelimb in 3-year-olds or older horses were significantly higher than in 2-year-olds. In contrast, the risk of SL desmitis in the forelimb at age 3 and 4 years was 2.23 and 2.11 times higher, respectively, than in 2-year-olds, but this increased to 5.07 times in those age ≥5 years. Entire males were at greater risk in comparison to females and geldings. Conclusions: The results suggest that the prevalence of SDF tendonitis and SL desmitis in the forelimb was associated with the horse's age and sex. The prevalence of SL desmitis increased further with age compared with SDF tendonitis, possibly reflecting a more rapid accumulation of degeneration in this structure. Potential relevance: The age-related risk demonstrated in this study provides further support that overstrain injuries are associated with accumulated degeneration. These data provide a valuable resource for further research into the aetiology of tendon injury in the racehorse.

Reasons for performing study: Injury to the superficial digital flexor tendon (SDFT) is common in competition horses. The SDFT contributes to locomotory efficiency by storing energy; such tendons have low safety margins. Tendons which merely position the limb, including the opposing common digital extensor tendon (CDET), are rarely injured. The current failure of strategies to prevent or effectively treat injury to the SDFT indicates the importance of understanding how it differs from tendons which are not injury-prone. Hypothesis: That the structural and material properties and matrix composition of the SDFT and CDET differ, reflecting their specific functional requirements in vivo. Methods: Forelimb tendons were harvested from 26 mature horses and loaded to failure prior to matrix composition analysis of specimens. Results: The SDFT had a significantly higher cross-sectional area, structural stiffness, failure load and failure strain and a lower elastic modulus than the CDET (P

Introduction Autologous mesenchymal stem cells (MSCs) are an attractive concept in regenerative medicine, but their mechanism of action remains poorly defined. No immune response is reported after in vivo injection of allogeneic equine MSCs or embryo-derived stem cells (ESCs) into the equine tendon, which may be due to the cells’ immune-privileged properties. This study further investigates these properties to determine their potential for clinical application in other tissues. Methods Mitomycin C-treated MSCs, ESCs, or differentiated ESCs (dESCs) were cultured with allogeneic equine peripheral blood mononuclear cells (PBMCs), and their effect on PBMC proliferation, in the presence or absence of interferon-gamma (IFN-γ) was determined. MSCs and super-antigen (sAg)-stimulated PBMCs were co-cultured directly or indirectly in transwells, and PBMC proliferation examined. Media from MSC culture were harvested and used for PBMC culture; subsequent PBMC proliferation and gene expression were evaluated and media assayed for IFN-γ, tumor necrosis factor alpha (TNF-α), and interleukin (IL)-10 and IL-6 proteins with enzyme-linked immunosorbent assay (ELISA). Results Co-culture of PBMCs with ESCs or dESCs did not affect baseline proliferation, whereas co-culture with MSCs significantly suppressed baseline proliferation. Stimulation of PBMC proliferation by using super-antigens (sAgs) was also suppressed by co-culture with MSCs. Inhibition was greatest with direct contact, but significant inhibition was produced in transwell culture and by using MSC-conditioned media, suggesting that soluble factors play a role in MSC-mediated immune suppression. The MSCs constitutively secrete IL-6, even in the absence of co-culture with PBMCs. MSC-conditioned media also brought about a change in the cytokine-expression profile of sAg-stimulated PBMCs, significantly reducing PBMC expression of IL-6, IFN-γ, and TNF-α. Conclusions Equine MSCs and ESCs possess a degree of innate immune privilege, and MSCs secrete soluble factors that suppress PBMC proliferation and alter cytokine expression. These properties may make possible the future clinical use of allogeneic stem cells to help standardize and broaden the scope of treatment of tissue injuries.

Dorsal carpal osteochondral injury is a major cause of lameness in horses undergoing high intensity training. Intra-articular corticosteroid treatment is used commonly to manage exercise-associated articular pain, but its use remains highly controversial in the equine athlete. This project, therefore, aimed to compare the mechanical properties of intra-articular MPA and diluent-treated middle carpal subchondral and cancellous bone in horses undergoing a short-term treadmill exercise programme. It was hypothesised that subchondral and cancellous bone mechanical properties are influenced by intra-articular administration of methylprednisolone acetate (MPA). Eight 2-year-old female horses had MPA or diluent administered into contralateral middle carpal joints at 14 day intervals, for a total of 4 treatments per horse. Horses underwent a standard treadmill exercise protocol until euthanasia (Day 70). Standard sites were located on the dorsal aspect of third, radial and intermediate carpal bones. Osteochondral samples from each test site were divided into subchondral bone and cancellous bone portions. These were dried, resin-embedded and gold-coated. Microhardness measurements were obtained at each test site. No significant effect of intra-articular treatment was detected. At each site, cancellous bone trabecular struts had an 18-19% higher microhardness value than the overlying subchondral bone. These findings indicate that intra-articular administration of MPA at this dose has no effect on subchondral or cancellous bone adaptation to short-term exercise and, therefore, on the propensity of carpal bones to injury. Further investigation into the calcified cartilage layer, effect of different corticosteroid preparations and diffusion of medication are required.

Specific tendons show a high incidence of partial central core rupture which is preceded by degeneration. In the performance horse, the superficial digital flexor tendon (SDFT) is most often affected. We have described previously the molecular changes that are associated with degeneration in the central core region of the equine SDFT. The pathophysiological mechanism leading to change in synthetic activity of central zone cells in degenerated tendons is not known. In this study, we test the hypothesis that ageing results in matrix composition changes within the central zone of the SDFT. Extracellular matrix composition and cellularity were analysed in equine SDFTs collected from Thoroughbred horses and compared with a flexor tendon which rarely shows degenerative change and subsequent injury (deep digital flexor tendon, DDFT). Data were examined for age-related changes to central and peripheral zone tissue of the SDFT and DDFT. Ageing in both tendons (SDFT and DDFT) resulted in a significant increase in collagen-linked fluorescence and a decrease in cellularity in the DDFT but not the SDFT. The central zone tissue from the SDFT had a significantly higher proportion of type III collagen than the peripheral zone of the tendon. The highest level of type III collagen was found in the central zone tissue of the SDFT from the older group of horses and this may represent the early stages of a degenerative change. Collagen content did not differ between the 2 flexor tendons; however, there were differences in collagen type and organisation. The SDFT had a higher type III collagen content, higher levels of the mature trifunctional collagen crosslink hydroxylysylpyridinoline, lower total chondroitin sulphate equivalent glycosaminoglycan content, smaller diameter collagen fibrils and a higher cellularity than the DDFT. In conclusion, differences in macromolecular composition exist between the flexor tendons and ageing contributes to a tendon specific change in composition.

Summary This study was performed to test the hypothesis that a localised bone hypertrophy could occur within the subchondral cancellous architecture of the third and radial carpal bones. Using 2 levels of controlled and defined exercise, it was observed that a high intensity treadmill exercise protocol resulted in functional adaptation of the carpal bones. The increase in trabecular thickening and density was seen to be localised to those regions underlying common sites of cartilage degradation, the interface of the thickened trabeculae with the normal architecture in the third carpal bone was coincident with a common site of clinical fractures. The bone changes were determined both qualitatively on examination of slab radiographs and quantified by dual energy x-ray absorptiometry. The findings from this study are relevant to mechanical factors involved in the pathophysiology of joint degeneration. The potential clinical implications of this study are in relation to changes in the type and duration of exercise regimens used in training of equine athletes. The rapid response of bone to mechanical stimulation has implications in the longer term for localised cartilage degradation. Imaging techniques could be developed to monitor these early bone changes in the specific areas identified in this study and thus allow appropriate changes in training intensity to minimise subsequent damage to the articular surface.

Injuries to the superficial digital flexor tendon are common in horses required to gallop and jump at speed. Partial rupture of this tendon usually occurs in the central core of the midmetacarpal region and may be preceded by localised degenerative changes. Post mortem examination of apparently normal equine flexor tendons has revealed an abnormal macroscopic appearance in the central core, characterised by a reddish discolouration. We have previously shown that there is also physical damage to the collagen fibres. In the present study we tested the hypothesis that the abnormal appearance is accompanied by changes in the composition of the extracellular matrix of the tendon. Biochemical analysis of the extracellular matrix demonstrated an increase in total sulphated glycosaminoglycan content, increase in the proportion of type III collagen and decrease in collagen linked fluorescence in the central core of 'degenerated' tendons relative to tissue from the peripheral region of the same tendon. Dry matter content and total collagen content were not significantly different between tendon zones or normal and 'degenerated' tendons. These changes suggest a change in cell metabolism and matrix turnover in the central core of the tendon and are likely to contribute to a decrease in mechanical properties in this part of the tendon, predisposing to the characteristic partial rupture of the tendon.

Summary Injury to the core of the mid-metacarpal region of the superficial digital flexor tendon in Thoroughbred racehorses is a very frequent but poorly understood condition. It has been suggested that subclinical changes induced by galloping exercise weaken the collagen in this region of the tendon, predisposing it to rupture. The longitudinally arranged collagen fibrils in tendon follow a planar waveform, termed the crimp. Fibril bundles with a smaller crimp angle fail at a lower level of strain than those with a larger crimp angle. This study tested the hypothesis that a specific 18 month exercise programme would result in significant reduction of collagen fibril crimp angle and period length in the core region of the superficial digital flexor tendon of young Thoroughbreds (21 ± 1 months), compared to the normal change in these parameters with age. Central region crimp angle and length were significantly lower in exercised horses than in control horses (P