Functional adaptation through changes in regional biochemical characteristics during maturation of equine superficial digital flexor tendons

OBJECTIVE: To quantify and compare biochemical characteristics of the extracellular matrix (ECM) of specimens harvested from tensional and compressive regions of the superficial digital flexor tendon (SDFT) of horses in age classes that include neonates to mature horses. SAMPLE POPULATION: Tendon specimens were collected on postmortem examination from 40 juvenile horses (0, 5, 12, and 36 months old) without macroscopically visible signs of tendonitis. PROCEDURE: Central core specimens of the SDFT were obtained with a 4-mm-diameter biopsy punch from 2 loaded sites, the central part of the mid-metacarpal region and the central part of the mid-sesamoid region. Biochemical characteristics of the collagenous ECM content (ie, collagen, hydroxylysylpyridinoline crosslink, and pentosidine crosslink concentrations and percentage of degraded collagen) and noncollagenous ECM content (percentage of water and glycosaminoglycans, DNA, and hyaluronic acid concentrations) were measured. RESULTS: The biochemical composition of equine SDFT was not homogeneous at birth with respect to DNA, glycosaminoglycans, and pentosidine concentrations. For most biochemical variables, the amounts present at birth were dissimilar to those found in mature horses. Fast and substantial changes in all components of the matrix occurred in the period of growth and development after birth. CONCLUSIONS AND CLINICAL RELEVANCE: Unlike cartilage, tendon tissue is not biochemically blank (ie, homogeneous) at birth. However, a process of functional adaptation occurs during maturation that changes the composition of equine SDFT from birth to maturity. Understanding of the maturation process of the juvenile equine SDFT may be useful in developing exercise programs that minimize tendon injuries later in life that result from overuse.     

Age-related changes to the molecular and cellular components of equine flexor tendons.

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.     

Influence of site and age on biochemical characteristics of the collagen network of equine articular cartilage

OBJECTIVE: To determine variations in biochemical characteristics of equine articular cartilage in relation to age and the degree of predisposition for osteochondral disease at a specific site. SAMPLE POPULATION: Articular cartilage specimens from 53 horses 4 to 30 years old. PROCEDURE: Healthy specimens were obtained from 2 locations on the proximal articular surface of the first phalanx that had different disease prevalences (site 1 at the mediodorsal margin and site 2 at the center of the medial cavity). Water, total collagen, and hydroxylysine contents and enzymatic (hydroxylysylpyridinoline [HP]) and nonenzymatic (pentosidine) crosslinking were determined at both sites. Differences between sites were analyzed by ANOVA (factors, site, and age), and age correlation was tested by Pearson's product-moment correlation analysis. Significance was set at P< 0.01. RESULTS: Correlation with age was not found for water, collagen, hydroxylysine contents, and enzymatic cross-linking. Nonenzymatic crosslinking was higher in older horses and was linearly related to age (r = 0.94). Water and collagen contents and HP and pentosidine crosslinks were significantly higher at site 1. Hydroxylysine content was significantly lower at site 1. CONCLUSIONS: Except for nonenzymatic glycation, the composition of articular cartilage collagen does not change significantly in adult horses. A significant topographic variation exists in biochemical characteristics of the articular cartilage collagen network in equine metacarpophalangeal joints. These differences may influence local biomechanical properties and, hence, susceptibility to osteochondral disease, as will greater pentosidine crosslinks in older horses that are likely to cause stiffer and more brittle cartilage.     

Control of the collagen fibril diameter in the equine superficial digital flexor tendon in horses by decorin

The distribution pattern of collagen fibril diameter in the equine superficial digital flexor tendon (SDFT) is known to differ in central and peripheral areas of some regions. This study reports the essence of collagen fibril differences among different regions of the equine SDFT by transmission electron microscopic (TEM) and high-voltage electron microscopic observations and biochemical analysis. The distribution of large collagen fibrils increased but the density of collagen fibrils decreased from the proximal metacarpal region to the distal metacarpal region. Large collagen fibrils with an irregular cross-sectional profile were found more frequently in the middle metacarpal region than in other regions. Three-dimensional reconstruction of images of irregularly shaped collagen fibrils revealed that these fibrils are formed through fusion of small collagen fibrils with large ones. The amount of decorin, which reportedly inhibits the lateral fusion of collagen fibrils, decreased in the direction of the distal metacarpal region. On the other hand, the size of decorin gradually increased in the direction of the distal metacarpal region. These results suggest that regional differences in collagen fibril distribution and density of collagen fibrils in the SDFT are due, at least in part, to fusion of collagen fibrils and the concomitant regional differences in the amount and size of decorin.     

A review of tendon injury: Why is the equine superficial digital flexor tendon most at risk?

Tendon injury is one of the most common causes of wastage in the performance horse; the majority of tendon injuries occur to the superficial digital flexor tendon (SDFT) whereas few occur to the common digital extensor tendon. This review outlines the epidemiology and aetiology of equine tendon injury, reviews the different functions of the tendons in the equine forelimb and suggests possible reasons for the high rate of failure of the SDFT. An understanding of the mechanisms leading to matrix degeneration and subsequent tendon gross failure is the key to developing appropriate treatment and preventative measures.     

Recombinant equine growth hormone does not affect the in vitro biomechanical properties of equine superficial digital flexor tendon

OBJECTIVE: To evaluate the effect of recombinant equine growth hormone (rEGH) on the in vitro biomechanical properties of normal adult equine superficial digital flexor tendon (SDFT). STUDY DESIGN: Completely randomized design. SAMPLE POPULATION: Nine Standardbred horses, 6 to 9 years of age with ultrasonographically normal forelimb SDFT. METHODS: Six horses were administered intramuscular (IM) rEGH at 10 microg/kg/day for 1 week, and then 20 microg/kg/day for another 5 weeks; 3 horses (control subjects) were administered an equivalent daily volume of sterile water IM. Horses were killed at the end of the 6-week treatment period, and both forelimb SDFT were harvested and stored at -70 degrees C. In vitro biomechanical testing was performed under uniaxial tension. Results were analyzed using a general linear model of analysis of variance; significance was set at P <.05. RESULTS: There were no differences in cross-sectional area, maximal load at failure, yield load, ultimate and yield tensile strain, ultimate and yield tensile stress, or stiffness between tendons from control and treated horses. CONCLUSIONS: Administration of rEGH to adult Standardbred horses for 6 weeks had no detectable effect on the in vitro biomechanical properties of normal SDFT. CLINICAL RELEVANCE: Administration of rEGH does not modulate the in vitro biomechanical properties of SDFT from adult Standardbred horses.     

The effect of recombinant equine growth hormone on the biomechanical properties of healing superficial digital flexor tendons in horses

OBJECTIVE: To evaluate the effect of recombinant equine growth hormone (rEGH) on the in vitro biomechanical properties of healing superficial digital flexor tendon (SDFT) in horses. STUDY DESIGN: Completely randomized design. SAMPLE POPULATION: Twelve Standardbred horses, 3 to 7 years of age, with ultrasonographically normal forelimb SDFT. METHODS: One week after induction of collagenase (2,000 U) induced superficial flexor tendonitis, horses were randomly divided into groups of 6. One group was administered intramuscular rEGH (10 microg/kg/day for 1 week, then 20 microg/kg/day for 5 weeks), whereas the other group (control subjects) were administered an equivalent volume of saline (0.9% NaCl) solution. At the end of this 6-week treatment, horses were killed and one forelimb SDFT from each horse was harvested for biomechanical testing under uniaxial tension. Results were analyzed using an unpaired Student's t test; significance was set at P 相似文献   

Effects of exercise on tenocyte cellularity and tenocyte nuclear morphology in immature and mature equine digital tendons

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.     

Functional adaptation of equine articular cartilage: the formation of regional biochemical characteristics up to age one year

Biochemical heterogeneity of cartilage within a joint is well known in mature individuals. It has recently been reported that heterogeneity for proteoglycan content and chondrocyte metabolism in sheep develops postnatally under the influence of loading. No data exist on the collagen network in general or on the specific situation in the horse. The objective of this study was to investigate the alterations in equine articular cartilage biochemistry that occur from birth up to age one year, testing the hypothesis that the molecular composition of equine cartilage matrix is uniform at birth and biochemical heterogeneity is formed postnatally. Water content, DNA content, glycosaminoglycan content (GAG) and biochemical characteristics of the collagen network (collagen content, hydroxylysine content and hydroxylysylpyridinoline [HP] crosslinks) were measured in immature articular cartilage of neonatal (n = 16), 5-month-old foals (n = 16) and yearlings (n = 16) at 2 predefined differently loaded sites within the metacarpophalangeal joint. Statistical differences between sites were analysed by ANOVA (P<0.01), and age correlation was tested by Pearson's product moment correlation analysis (P<0.01). In neonatal cartilage no significant site differences were found for any of the measured biochemical parameters. This revealed that the horse has a biochemically uniform joint (i.e. the cartilage) at birth. In the 5-month-old foals and yearlings, significant site differences, comparable to those in the mature horse, were found for DNA, GAG, collagen content and hydroxylysine content. This indicates that functional adaptation of articular cartilage to weight bearing for these biochemical parameters takes place during the first months postpartum. Water content and HP crosslinks showed no difference between the 2 sites from neonatal horses, 5-month-old animals and yearlings. At both sites water, DNA and GAG decreased during maturation while collagen content, hydroxylysine content and HP crosslinks increased. We propose that a foal is born with a uniform biochemical composition of cartilage in which the functional adaptation to weight bearing takes place early in life. This adaptation results in biochemical and therefore biomechanical heterogeneity and is thought to be essential to resist the different loading conditions to which articular cartilage is subjected during later life. As collagen turnover is extremely low at mature age, an undisturbed functional adaptation of the collagen network of articular cartilage at a young age may be of significant importance for future strength and resistance to injury.     

Age-related morphometry of equine calcified cartilage

Although there are many studies in the equine literature focused on articular diseases and the aetiology of osteoarthritis, few have concentrated on normal articular structures and how they change with age. The objective of this investigation was to study the thickness and morphology of the calcified cartilage layer of the distal metacarpus over a range of ages. A parasagittal slab of bone was sectioned from the region of sesamoid contact on the medial condyle of the metacarpi from 34 horses. The slab of bone was preserved, dehydrated and embedded, undecalcified, in methylmethacrylate and then stained with toluidine blue. Six repeatable fields of interest from the distal aspect of each metacarpus were digitised and examined to determine the morphology of the calcified cartilage layer. The thickness of the calcified cartilage, range 88-426 microm, was estimated using a method of integration. The results indicate an age-related influence on the thickness of the calcified cartilage layer, generally increased in older horses. While this finding is significant, perhaps more importantly a positional relationship was also identified, indicating that pressures endured by different regions within a joint may dictate morphological development of the tissues. This study has begun to lay the groundwork to determine whether the calcified layer of the hyaline cartilage could be involved in the development of osteoarthritis.     

Topographical mapping of biochemical properties of articular cartilage in the equine fetlock joint

The aim of this study was to evaluate topographical differences in the biochemical composition of the extracellular matrix of articular cartilage of the normal equine fetlock joint. Water content, DNA content, glycosaminoglycan (GAG) content and a number of characteristics of the collagen network (total collagen content, levels of hydroxylysine- (Hyl) and the crosslink hydroxylysylpyridinoline, (HP) of articular cartilage in the proximal 1st phalanx (P1), distal 3rd metacarpal bone (MC), and proximal sesamoid bones (PSB) were determined in the left and right fetlock joint of 6 mature horses (age 5-9 years). Twenty-eight sites were sampled per joint, which included the clinically important areas often associated with pathology. Biochemical differences were evaluated between sampling sites and related with the predisposition for osteochondral injury and type of loading. Significant regional differences in the composition of the extracellular matrix existed within the joint. Furthermore, left and right joints exhibited biochemical differences. Typical topographic distribution patterns were observed for each parameter. In P1 the dorsal and palmar articular margin showed a significantly lower GAG content than the more centrally located sites. Collagen content and HP crosslinks were higher at the joint margins than in the central area. Also, in the MC, GAG content was significantly lower at the (dorsal) articular margin compared with the central area. Consistent with findings in P1, collagen and HP crosslinks were significantly lower in the central area compared to the (dorsal) articular margin. Biochemical and biomechanical heterogeneity of articular cartilage is supposed to reflect the different functional demands made at different sites. In the present study, GAG content was highest in the constantly loaded central areas of the joint surfaces. In contrast, collagen content and HP crosslinks were higher in areas intermittently subjected to peak loading which suggests that the response to a certain type of loading of the various components of the extracellular matrix of articular cartilage are different. The differences in biochemical characteristics between the various sites may help to explain the site specificity of osteochondral lesions commonly found in the equine fetlock joint. Finally, these findings emphasise that the choice of sampling sites may profoundly influence the outcome of biochemical studies of articular cartilage.     

MR IMAGING FEATURES OF SURGICALLY INDUCED CORE LESIONS IN THE EQUINE SUPERFICIAL DIGITAL FLEXOR TENDON

Tendon injuries are common in athletic humans and horses. Ultrasonography is the diagnostic method of choice in horses with tendon injuries but there is increasing application of magnetic resonance (MR) imaging to monitor and follow‐up tendon healing. A core lesion was created in the superficial digital flexor tendon (SDFT) of each forelimb of four horses. One of the four horses was euthanized at 2, 4, 8, and 12 weeks after creation of the lesion. MR examinations of the SDFT were performed immediately post mortem in a 1.5 T Siemens Symphony magnet and compared with histologic findings. Measurements from the MR images were also compared to ultrasonographic measurements available from the same lesions. Tendon lesions appeared as well‐circumscribed hyperintensities in the core of the SDFT on all pulse sequences. Lesions were most conspicuous on fat‐suppressed fast low angle shot (FLASH) sequences and least conspicuous on T2 transverse dual turbo spin echo (T2 TSE) sequences. The signal‐difference‐to‐noise ratio decreased with the age of the lesion in all sequences in this study. Twelve‐week‐old lesions were not visible on T2 TSE images but in all other sequences the lesion remained hyperintense. The lesion volume and maximum cross‐sectional area of core lesions were significantly smaller in T2 TSE images than in other MR sequences. The lesion volume and maximum cross‐sectional area of core lesions were significantly larger in proton density, T1, and FLASH sequences and significantly smaller in T2 sequences than when measured from ultrasonographic images. Through comparison between sequences, MR imaging may be able to provide information on various stages of tendon healing.     

Microwave thermography: a non-invasive technique for investigation of injury of the superficial digital flexor tendon in the horse.

Microwave thermographs were recorded from 77 normal horses. In 51% the lowest temperature was recorded in the mid-metacarpal region, and in 41% it was in the distal metacarpal region. The mean temperature of the normal limbs ranged from 25.04 to 37.4 degrees C. Maximum temperature differences between symmetrical points in both forelimbs ranged from 0 to 5.33 degrees C and differences in mean limb temperatures between both forelimbs ranged from 0 to 2.91 degrees C. In 48 horses with acute (less than 4 weeks' duration) injury of the superficial digital flexor tendon (SDFT) (36 unilateral, 12 bilateral) and 12 horses with acute injury of the soft tissues of the palmar metacarpal region other than the SDFT (all unilateral) 66% of forelimbs had acute SDFT injury, and 50% of those with other soft tissue injuries, had elevations of the temperature in the mid- or distal metacarpal region. Abnormal values for mean limb temperature, difference in mean limb temperature and maximum temperature difference between locations in opposite forelimbs were detected in 75% of the horses with SDFT injury and in only 16% of the horses with other soft tissue injury. The sensitivity of microwave thermography for the detection of SDFT injury was 81% and the specificity 74%. When 30 horses in National Hunt training were examined weekly for 5 weeks, 2 horses sustained SDFT injury during that period. The microwave thermographs recorded from these 2 horses, at 1 and 2 weeks before the onset of clinical signs, were abnormal. However, 16 horses which did not develop tendon injury also displayed thermographic abnormalities.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 4. Morphometric, microscopic and biomechanical properties of the digital tendons of the forelimb

AIM: To determine the weight, volume, density and cross-sectional area (CSA) of the digital flexor tendons, common digital extensor tendon (CDET) and suspensory ligament (SL) of the forelimb of young Thoroughbred horses in early training, and to assess the response to a training programme of known parameters of superficial digital flexor tendon (SDFT) tissue at mid-metacarpal level. METHODS: The tendons of seven 2-year-old Thoroughbred horses in training were inspected, transected into segments of known length, and compared with those from seven untrained horses matched for age, sex and breed. The weight, volume, density and CSA of each segment, and the crimp angle, histological features, and biomechanical ultimate stress and stiffness of tendon samples from the mid-metacarpal region of the SDFT were determined. RESULTS: There was no macroscopic evidence of swelling or discolouration in any of the tendon segments or cut surfaces. The volumes of SDFT and CDET segments of horses in the trained group were significantly greater than those in the untrained group (p=0.036 and p=0.039, respectively). A greater increase in volume than weight resulted in a lower density in the SDFT but not CDET in trained compared with untrained horses (p=0.038). CSA of these two tendons was significantly greater in the trained group (p=0.002 and 0.036, respectively), the percentage increase being greater in the CDET than the SDFT. The number of tenocytes at four sites in the mid-metacarpal SDFT region was less in trained than untrained horses (p=0.025). There was no histological evidence of inflammation, and no difference in crimp angle between groups. There was no significant between-group difference in stiffness or ultimate stress of tendon strips. CONCLUSIONS: Volume and CSA of the SDFT and CDET were larger in trained than untrained horses. The SDFT was less dense in the trained group. Because no evidence of tendonitis was detected and training appeared to have no significant effect on crimp angle or biomechanical properties of tendon strips, the size and density changes were presumed to be adaptive and induced by the training. CLINICAL RELEVANCE: Although evident in this in vitro study, the detection of adaptive from initial pathological increase in size of the SDFT is likely to be difficult in vivo.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 4. Morphometric,microscopic and biomechanical properties of the digital tendons of the forelimb

AIM: To determine the weight, volume, density and cross-sectional area (CSA) of the digital flexor tendons, common digital extensor tendon (CDET) and suspensory ligament (SL) of the forelimb of young Thoroughbred horses in early training, and to assess the response to a training programme of known parameters of superficial digital flexor tendon (SDFT) tissue at mid-metacarpal level.

METHODS: The tendons of seven 2-year-old Thoroughbred horses in training were inspected, transected into segments of known length, and compared with those from seven untrained horses matched for age, sex and breed. The weight, volume, density and CSA of each segment, and the crimp angle, histological features, and biomechanical ultimate stress and stiffness of tendon samples from the mid-metacarpal region of the SDFT were determined.

RESULTS: There was no macroscopic evidence of swelling or discolouration in any of the tendon segments or cut surfaces. The volumes of SDFT and CDET segments of horses in the trained group were significantly greater than those in the untrained group (p=0.036 and p=0.039, respectively). A greater increase in volume than weight resulted in a lower density in the SDFT but not CDET in trained compared with untrained horses (p=0.038). CSA of these two tendons was significantly greater in the trained group (p=0.002 and 0.036, respectively), the percentage increase being greater in the CDET than the SDFT. The number of tenocytes at four sites in the mid-metacarpal SDFT region was less in trained than untrained horses (p=0.025). There was no histological evidence of inflammation, and no difference in crimp angle between groups. There was no significant between-group difference in stiffness or ultimate stress of tendon strips.

CONCLUSIONS: Volume and CSA of the SDFT and CDET were larger in trained than untrained horses. The SDFT was less dense in the trained group. Because no evidence of tendonitis was detected and training appeared to have no significant effect on crimp angle or biomechanical properties of tendon strips, the size and density changes were presumed to be adaptive and induced by the training.

CLINICAL RELEVANCE: Although evident in this in vitro study, the detection of adaptive from initial pathological increase in size of the SDFT is likely to be difficult in vivo.     

Characterization and clinical application of mesenchymal stem cells from equine umbilical cord blood

Tendinitis of the superficial digital flexor tendon (SDFT) is a significant cause of lameness in horses; however, recent studies have shown that stem cells could be useful in veterinary regenerative medicine. Therefore, we isolated and characterized equine umbilical cord blood mesenchymal stem cells (eUCB-MSCs) from equine umbilical cord blood obtained from thoroughbred mares during the foaling period. Horses that had tendinitis of the SDFT were treated with eUCB-MSCs to confirm the therapeutic effect. After eUCB-MSCs transplantation, the core lesion in the SDFT was found to decrease. These results suggest that transplantation using eUCB-MSCs could be another source of cell treatment.     

Expression of bone morphogenetic protein-6 and -2 and a bone morphogenetic protein antagonist in horses with naturally acquired osteochondrosis

OBJECTIVE: To determine the mRNA expression of bone morphogenetic protein (BMP)-6 and -2 and a BMP antagonist (Noggin) in horses with osteochondrosis. SAMPLE POPULATION: Samples of articular cartilage from affected stifle or shoulder joints of 10 immature horses with naturally acquired osteochondrosis and corresponding joints of 9 clinically normal horses of similar age; additionally, samples of distal femoral growth plate cartilage and distal femoral articular cartilage were obtained from a normal equine fetus. PROCEDURE: Cartilage specimens were snap-frozen in liquid nitrogen, and total RNA was isolated. Adjacent specimens were fixed in 4% paraformaldehyde for histologic examination. Expression of BMP-6, BMP-2, and Noggin mRNA was evaluated by real-time quantitative polymerase chain reaction (PCR) assays. Spatial tissue mRNA expression of BMP-6 was determined by in situ hybridization. RESULTS: Nucleotide sequences were obtained for portions of the BMP-6 propeptide and mature peptide region, as well as the signal and mature peptide region of Noggin. Expression of BMP-6, BMP-2, and Noggin mRNA was found to be similar in cartilage from normal and osteochondrosis-affected horses. Spatial expression of BMP-6 correlated with the middle and deep layers of articular cartilage; no differences were observed in overall expression between cartilage specimens from the 2 groups of horses. No expression of BMP-6 was found in the superficial layer, subchondral bone, or osteochondrosis-affected cleft fibrous tissue. CONCLUSIONS AND CLINICAL RELEVANCE: Although these signaling peptides may play important roles in cartilage differentiation, results did not provide evidence to suggest that they are involved in the disease process of osteochondrosis.     

Biochemical analysis of the articular cartilage and subchondral and trabecular bone of the metacarpophalangeal joint of horses with early osteoarthritis

OBJECTIVE: To assess whether site-related changes in biochemical composition are present in the cartilage and subchondral and trabecular bone of the metacarpophalangeal joint of horses with early osteoarthritis. SAMPLE POPULATION: Right metacarpophalangeal joints from 59 mature warmblood horses. PROCEDURE: Biochemical data (cross-link, amino acid, DNA, and ash contents; denatured collagen and glycosaminoglycan [GAG] concentrations; bone mineral density; and mineral composition) were obtained from 2 differently loaded sites of phalanx I cartilage and subchondral and trabecular bone samples; data were compared with previously published values from nonosteoarthritic equine joints. RESULTS: Compared with findings in nonosteoarthritic joints, GAG concentration was lower in cartilage from osteoarthritic joints and there was a loss of site differences in cellularity and lysylpyridinoline (LP) cross-link content. In subchondral bone, LP cross-link content was decreased overall and there was a loss of site differences in osteoarthritic joints; ash content was higher in the osteoarthritic joints. Hydroxyproline content in trabecular bone from osteoarthritic joints was greater than that in nonosteoarthritic trabecular bone. In all 3 layers and at both sites, the linear increase of the pentosidine cross-link content with age had diminished or was not apparent in the horses with osteoarthritic joints. CONCLUSIONS AND CLINICAL RELEVANCE: In equine metacarpophalangeal joints with early osteoarthritis, distinct biochemical changes were detected in the cartilage and subchondral and trabecular bone. The dissimilarity in response of the different tissues and differences between the sites that are affected may be related to differences in biomechanical loading and transmission and dissipation of force.     

Biochemical characterisation of navicular hyaline cartilage,navicular fibrocartilage and the deep digital flexor tendon in horses with navicular disease

The study hypothesis was that navicular disease is a process analogous to degenerative joint disease, which leads to changes in navicular fibrocartilage and in deep digital flexor tendon (DDFT) matrix composition and that the process extends to the adjacent distal interphalangeal joint. The objectives were to compare the biochemical composition of the navicular articular and palmar cartilages from 18 horses with navicular disease with 49 horses with no history of front limb lameness, and to compare navicular fibrocartilage with medial meniscus of the stifle and collateral cartilage of the hoof. Cartilage oligomeric matrix protein (COMP), deoxyribonucleic acid (DNA), total glycosaminoglycan (GAG), metalloproteinases MMP-2 and MMP-9 and water content in tissues were measured. Hyaline cartilage had the highest content of COMP and COMP content in hyaline cartilage and tendon was higher in lame horses than in sound horses (p<0.05). The concentration of MMP-2 amount in hyaline cartilage was higher in lame horses than in sound horses. The MMP-2 amounts were significantly higher in tendons compared to other tissue types. Overall, 79% of the lame horses with lesions had MMP-9 in their tendons and the amount was higher than in sound horses (p<0.05). In horses with navicular disease there were matrix changes in navicular hyaline and fibrocartilage as well as the DDFT with potential implications for the pathogenesis and management of the condition.     

Development of biochemical heterogeneity of articular cartilage: influences of age and exercise

The objective of this study was to document the development of biochemical heterogeneity from birth to maturity in equine articular cartilage, and to test the hypothesis that the amount of exercise during early life may influence this process. Neonatal foals showed no biochemical heterogeneity whatsoever, in contrast to a clear biochemical heterogeneity in mature horses. The process of formation of site differences was almost completed in exercised foals age 5 months, but was delayed in those deprived of exercise. For some collagen-related parameters, this delay was not compensated for after an additional 6 month period of moderate exercise. It is concluded that the functional adaptation of articular cartilage, as reflected in the formation of biochemical heterogeneity in the horse, occurs for the most part during the first 5 months postpartum. A certain level of exercise seems essential for this process and withholding exercise in early life, may result in a delay in the adaptation of the cartilage.