Associations of horse age, joint type, and osteochondral injury with serum and synovial fluid concentrations of type II collagen biomarkers in Thoroughbreds.

OBJECTIVE: To determine the effects of horse age, osteochondral injury, and joint type on a synthesis biomarker and 3 degradative biomarkers of type II collagen in Thoroughbreds. ANIMALS: Healthy rested adult (3- to 12-year-old) Thoroughbreds (n = 19), yearling (1- to 2-year-old) Thoroughbreds (40), and Thoroughbred racehorses (2 to 7 years old) undergoing arthroscopic surgery for removal of osteochondral fragments that resulted from training or racing (41). PROCEDURES: Samples of blood and metacarpophalangeal, metatarsophalangeal, or carpal joint synovial fluid (SF) were collected from all horses. Commercially available assays were used to analyze SF and serum concentrations of type II collagen biomarkers of synthesis (carboxy propeptide of type II collagen [CPII]) and degradation (cross-linked C-telopeptide fragments of type II collagen [CTX II], neoepitope generated by collagenase cleavage of type I and II collagen [C1,2C], and neoepitope generated by collagenase cleavage of type II collagen [C2C]). RESULTS: Osteochondral injury affected concentrations of CPII, CTX II, C1,2C, and C2C in SF, serum, or both, compared with concentrations in healthy adult horses. Compared with adult horses, yearling horses had increased SF or serum concentrations of degradative biomarkers (CTX II, C1,2C, and C2C). Concentrations were higher in carpal than metacarpophalangeal or metatarsophalangeal joints for all biomarkers in osteochondral-injured horses. Variable differences in SF concentrations between joint types were detected in healthy adult and yearling horses. CONCLUSIONS AND CLINICAL RELEVANCE: Horse age, osteochondral injury, and joint type all significantly affected type II collagen biomarker concentrations in SF and serum of Thoroughbreds.

Cartilage thickness and split-line pattern at the canine humeral trochlea.

OBJECTIVE: To characterise the humeral trochlea in middle to large breed dogs in respect to split-line pattern and cartilage thickness. METHODS: In 15 paired cadaveric elbow joints of mature dogs (>20 kg body weight) collagen network orientation of the hyaline cartilage of the humeral trochlea was visualised using a traditional split-line technique in which a dissecting needle dipped in India ink was inserted into the cartilage (n = 10). Cartilage thickness was measured radiographically on osteochondral plugs harvested at four representative locations within the joint surface of the humeral trochlea (n = 15). RESULTS: The joint surface of the humeral trochlea showed a distinct pattern of centripetally oriented split-lines with less pronounced or even absent split-lines caudo- proximally towards the olecranon fossa. Median cartilage thickness at the canine humeral trochlea was 0.51 mm (interquartile range: 0.42 - 0.61 mm). Centrally, at the region where osteochondrosis lesions commonly occur in middle to large breed dogs, the median cartilage thickness was 0.55 mm (interquartile range: 0.48 - 0.62 mm). CLINICAL SIGNIFICANCE: When focusing on anatomical joint resurfacing while performing osteochondral transplantation at the canine humeral trochlea, transplants should be implanted such that their split-lines are oriented centripetally. Hyaline cartilage thickness of transferred grafts should be in the range of half a millimetre to optimally match the situation at the canine humeral trochlea.

Interleukin-6 and high mobility group box protein-1 in synovial membranes and osteochondral fragments in equine osteoarthritis.

Cytokine production in synovial membranes (SM) and osteochondral fragments (OCF) may influence the development of equine osteoarthritis (OA). In this study, the presence of interleukin (IL)-6 and cytoplasmic and extracellular high mobility group box protein (HMGB)-1 in SM and osteochondral tissue from healthy and diseased equine joints was investigated by immunohistochemistry. Additionally, microscopic synovitis was graded. IL-6 was commonly found in SM cells and in chondrocytes in uncalcified cartilage of OCF, whereas little staining was detected in healthy cartilage. Cytoplasmic and/or extracellular HMGB-1 was widespread only in SM from diseased joints, and also detected in OCF in areas of cartilage damage, fibrous repair tissue, and tidemark reduplication. Joints with OCF and cartilage lesions (without OCF) showed significantly higher median synovitis scores than healthy joints (p=0.013 and p=0.042, respectively). The study identifies OCF as a source of inflammatory mediators in equine OA, as shown by the presence of IL-6 and extracellular HMGB-1 in the fragment. Based upon HMGB-1 release in SM and OCF, further studies to investigate possible involvement of HMGB-1 in the pathogenesis of OA are warranted.

Micro-computed tomography of early lesions of osteochondrosis in the tarsus of foals.

INTRODUCTION: Osteochondrosis (OC) is an important developmental orthopedic disease of human and equine patients. The disease is defined as a focal disturbance in enchondral ossification. In horses, the disturbance can occur secondary to failure of the blood supply to growth cartilage. Diagnosis of the early, subclinical stages that can clarify the etiology is currently confined to cross-sectional histological examination. The potential for micro-computed tomography (micro-CT) with angiography to detect early lesions of OC has not yet been investigated. MATERIALS AND METHODS: Nine Standardbred foals bred from parents with OC of the tarso-crural joint were sacrificed at weekly intervals from birth to 7 weeks of age. Permanent barium angiograms were created within one hind limb post mortem, and samples collected from two predilection sites for OC within the tarso-crural joint of the perfused hind limb. The resulting 18 sample blocks were scanned with a custom-built micro-CT equipment set-up, and analyzed as 2D slices and 3D volume rendered models before sectioning for conventional histological examination. RESULTS: Histological examination identified eight early lesions in seven locations within six joints from the nine foals. Micro-CT with angiography was able to detect seven lesions in the same sites as histological examination. Lesions consisted of non-perfused foci within growth cartilage. No perfused vessels exited from subchondral bone deep to any lesion. Six of the seven lesions were associated with focal defects in the subchondral bone plate. Evidence of ongoing ossification was seen in three out of the seven lesions and included one separate center of ossification. CONCLUSION: Micro-CT was a useful technique for examination of early lesions of OC. The results of micro-CT were compatible with failure of cartilage canal vessels at the point where they cross the ossification front. Resultant areas of ischemic chondronecrosis were associated with focal delay in enchondral ossification as visualized in 3D volume rendered models. Micro-CT combined with histology clarified the role of different forms of ossification in the secondary repair responses to lesions.

MT3-MMP (MMP-16) is downregulated by in vitro cytokine stimulation of cartilage, but unaltered in naturally occurring equine osteoarthritis and osteochondrosis.

Matrix degradation by metalloproteinases is considered a key feature in the loss of articular cartilage seen in many joint diseases. Membrane-type matrix metalloproteinase-3 (MT3-MMP) expression is elevated in human cartilage in end-stage osteoarthritis. We investigated whether MT3-MMP is similarly regulated in cartilage in two naturally occurring arthropathies in vivo and whether proinflammatory cytokines regulate its expression in vitro. MT3-MMP expression was evaluated in cartilage from horses with osteoarthritis and osteochondrosis and compared with age- and site-matched normal cartilage. MT3-MMP also was measured in normal cartilage stimulated with proinflammatory cytokines. MT3-MMP expression was not significantly altered in either osteoarthritis or osteochondrosis cartilage. However, gene expression was significantly downregulated by the addition of recombinant human interleukin-1beta, oncostatin M, or tumor necrosis factor-alpha to normal cartilage explants. The results suggest that MT3-MMP may not have a role in matrix destruction in equine cartilage diseases. Further work is required to characterize its regulation and function.

Associations between candidate gene markers at a quantitative trait locus on equine chromosome 4 responsible for osteochondrosis dissecans in fetlock joints of South German Coldblood horses.

A previously accomplished whole-genome scan for osteochondrosis (OC) and OC dissecans (OCD) in South German Coldblood horses using 250 microsatellite markers identified putative quantitative trait loci (QTL). A chromosome-wide significant QTL for fetlock OCD was found on Equus caballus chromosome (ECA) 4q at a relative position of 70.0-73.3 cM. The aim of this study was to analyze associations of single nucleotide polymorphisms (SNPs) in candidate genes for OC in this region. The association analysis included 32 affected and 64 unaffected horses. Three SNPs located in intron 8, intron 9, and 3'-untranslated region (UTR) of the acyloxyacyl hydrolase (AOAH) gene on ECA4q were significantly associated with OCD in fetlock joints. In order to control for systematic environmental and quantitative genetic effects, we employed a linear animal model. The association of the SNP (AJ543065:g.703A>G) in the 3'-UTR of exon 21 was confirmed in the animal model analysis and a significant additive genetic effect for fetlock OCD of 0.42 (P = 0.002) and a dominance effect of -0.32 (P = 0.03) was estimated. This is the first report on a marker in population-wide linkage disequilibrium with equine OCD in fetlock joints.

Epiphyseal cartilage canal blood supply to the distal femur of foals.

REASONS FOR PERFORMING STUDY: The developmental pattern of the cartilage canal blood supply to epiphyseal growth cartilage has been linked to osteochondrosis (OC) in the tarsus of foals. This pattern has not yet been described in the distal femur, another site frequently affected by OC. OBJECTIVE: To describe the developmental pattern of the blood supply to the distal femoral epiphyseal growth cartilage in 8 Standardbred foals age 0-7 weeks. METHODS: One foal was sacrificed weekly from birth to age 7 weeks (n=8) to undergo a barium perfusion procedure to demonstrate vessels within cartilage canals of one hindlimb. The distal end of the femur was cleared in methyl salicylate and perfused vessels were studied in the intact bones. Each distal femur was then sawed into 5 mm thick slabs in the transverse plane, and the slabs decalcified and radiographed. Finally, the lateral trochlear ridge was separated from each slab and examined histologically. RESULTS: The cartilage canal blood supply regressed with increasing age, but several regions remained vascularised in the oldest foal at age 7 weeks. Vessels arose from perichondrial and subchondral arterial sources, and coursed perpendicular or parallel to the ossification front. The midsection of parallel vessels became incorporated into the ossification front during growth. Anastomoses formed and vessels within the distal portion of canals with an original perichondrial source shifted to use subchondral vessels as their arterial source. Both parallel and perpendicular vessels therefore traversed the ossification front to enter cartilage canals. No histological lesions were observed in sections from any of the foals. CONCLUSION: The same anatomical feature (traversing the ossification front to enter cartilage canals) reported to render vessels vulnerable to failure in the tarsus was also present in the distal femur of foals. POTENTIAL RELEVANCE: OC may occur by the same pathogenetic mechanism in the distal femur as in the tarsus of foals.

Early changes in biomarkers of skeletal metabolism and their association to the occurrence of osteochondrosis (OC) in the horse.

REASON FOR PERFORMING STUDY: Diagnosis of osteochondrosis (OC) is based on clinical signs and radiography, but alternative methods for detection at an early stage would be useful. OBJECTIVES: To determine in the juvenile horse the relationship between serum concentrations of a number of biomarkers that reflect changes in cartilage and bone turnover and age, feeding level, growth, and the occurrence of OC. METHODS: Foals were assigned to a high (n = 20) or moderate (n = 19) feeding level group from birth to age 1 year. Bodyweight, withers height and cannon width were measured. Osteoarticular status was assessed radiographically at 5.5 and 11 months in all foals, and by necropsy at 12 months for 8 foals/group. Serum biomarkers of bone (osteocalcin, CTX-1) and cartilage (CPII, C2C) metabolism were assayed at 8 time points between ages 2 and 52 weeks. Ratios between biomarkers of tissue formation and degradation were calculated at each time point. RESULTS: Consistent age-related patterns in biomarker serum concentrates were found, indicating a markedly higher metabolism before age 20 weeks but concentrations were not affected by feeding level. Bodyweight was correlated negatively to C2C and CTX-1, and withers height was positively correlated to osteocalcin and the osteocalcin/CTX-1 and CPII/ C2C ratios. Osteocalcin concentration at 2 weeks and CPII/ C2C ratio at 20 weeks had strong positive correlations to OC, as diagnosed radiographically at 5.5 months. Osteocalcin had a strong correlation with radiographically detected OC at 11 months but at that time there was no significant relationship between CPII/C2C ratio and OC. CONCLUSIONS: Occurrence of OC lesions is significantly associated with anabolic changes in bone metabolism during the first weeks post partum, given the strong relation with osteocalcin. POTENTIAL RELEVANCE: Measuring osteocalcin concentrations during the first few weeks post partum may have potential value for the prediction of risk for OC development.

Epiphyseal cartilage canal blood supply to the tarsus of foals and relationship to osteochondrosis.

REASON FOR PERFORMING STUDY: Pathological changes in the blood supply to growth cartilage have been implicated in the pathogenesis of osteochondrosis (OC) in horses, but have not been reported using vascular perfusion techniques. OBJECTIVE: To describe the developmental pattern of cartilage canal vessels in the distal tibial epiphysis and talar growth cartilage of foals. METHODS: Nine foals bred from parents with OC were sacrificed between the ages of 0 and 7 weeks to undergo a barium perfusion procedure. The distal end of the tibia and the entire talus were cleared in methyl salicylate and perfused vessels studied in the intact bones. Slabs with a thickness of 4-5 mm from 3 predilection sites for OC were examined in the stereomicroscope and with light microscopy. RESULTS: Cartilage canals were present for a limited period of growth. Perfused vessels initially entered canals from the perichondrium. Vessels in the proximal portion of canals retained their perichondrial arterial source throughout. With time, the ossification front advanced to incorporate the mid-portion of canals; and anastomoses formed between canal vessels and subchondral vessels. A shift occurred and vessels in the distal terminus of canals came to use subchondral vessels as their arterial source. Twelve histological lesions were found in 7 foals. All contained necrotic vessels surrounded by necrotic growth cartilage and 3 caused macroscopically visible delay in endochondral ossification. Lesions were located where vessels traversed the ossification front to enter the distal terminus of canals. CONCLUSION: Cartilage canal vessels are particularly susceptible to failure at the point where they cross the ossification front, with consequences for the viability of those chondrocytes that depend on them. POTENTIAL RELEVANCE: A better understanding of how lesions of OC arise may improve the ability to identify, monitor, prevent and treat this disorder. Involvement of cartilage canals in the pathogenesis of equine tarsal OC plausibly explains several clinical features of this disease.

Cartilage matrix changes in the developing epiphysis: early events on the pathway to equine osteochondrosis?

REASONS FOR PERFORMING STUDY: The earliest osteochondrosis (OC) microscopic lesion reported in the literature was present in the femorotibial joint of a 2-day-old foal suggesting that OC lesions and factors initiating them may arise prior to birth. OBJECTIVE: To examine the developing equine epiphysis to detect histological changes that could be precursors to OC lesions. METHODS: Osteochondral samples from 21 equine fetuses and 13 foals were harvested from selected sites in the scapulohumeral, humeroradial, metacarpophalangeal, femoropatellar, femorotibial, tarsocrural and metatarsophalangeal joints. Sections were stained with safranin O and picrosiruis red to assess cartilage changes and structural arrangement of the collagen matrix. RESULTS: Extracellular matrix changes observed included perivascular areas of paleness of the proteoglycan matrix associated with hypocellularity and, sometimes, necrotic chondrocytes. These changes were most abundant in the youngest fetuses and in the femoropatellar/femorotibial (FP/FT) joints. Indentations of the ossification front were also observed in most specimens, but, most frequently, in scapulohumeral and FP/FT joints. A cartilage canal was almost always present in these indentations. The vascular density of the cartilage was higher in the youngest fetuses. In these fetuses, the most vascularised joints were the metacarpo- and metatarsophalangeal joints but their cartilage canals regressed quickly. After birth, the most vascularised cartilage was present in the FP/FT joint. Articular cartilage differentiated into 4 zones early in fetal life and the epiphyseal cartilage also had a distinct zonal cartilage structure. A striking difference was observed in the collagen structure at the junction of the proliferative and hypertrophic zones where OCD lesions occur. CONCLUSION: Matrix and ossification front changes were frequently observed and significantly associated with cartilage canals suggesting that they may be physiological changes associated with matrix remodelling and development. The collagen structure was variable through the growing epiphysis and a differential in biomechanical properties at focal sites may predispose them to injury.

Effect of dietary nutrients on osteochondrosis lesions and cartilage properties in pigs.

OBJECTIVE: To evaluate dietary ingredients involved in cartilage and bone metabolism and their influence on osteochondrosis lesions in swine. ANIMALS: 80 crossbred gilts (mean initial weight, 39 kg). PROCEDURES: Pigs (10 pigs/treatment) were fed a corn-soybean meal basal (control) diet or the basal diet supplemented with additional minerals (copper and manganese or silicon), amino acids (proline and glycine; a combination of leucine, isoleucine, and valine; or methionine and threonine), or fatty acids (provided by fish oil) for 84 days. Pigs were then slaughtered and the distal portion of the left femur was collected for determination of osteochondrosis lesions at the femoral condyle. After evaluation of external joint surfaces, the distal portion of the femur was sectioned to evaluate lesions in the growth plate and articular cartilage. Additionally, a cartilage specimen was obtained from the patella for analysis. RESULTS: Pigs fed diets containing high amounts of methionine and threonine or the diet containing all additional ingredients had significantly lower total severity scores, compared with scores for pigs fed the control diet or a diet supplemented with fish oil. Pigs fed diets containing additional proline and glycine, copper and manganese, methionine and threonine, or all additional ingredients had significantly lower overall scores, compared with scores for pigs fed the control diet or a diet supplemented with fish oil. CONCLUSIONS AND CLINICAL RELEVANCE: Dietary manipulation decreased the severity of osteochondrosis lesions, compared with results for pigs fed a control diet. However, additional research on optimal concentrations and combinations of dietary components is needed.

Autogenous osteochondral grafting for treatment of stifle osteochondrosis in dogs.

OBJECTIVE: To develop and assess clinical outcomes for osteochondral autografting for treatment of stifle osteochondrosis (OC) in dogs. STUDY DESIGN: Retrospective case series. ANIMALS: Dogs with stifle OC (n=10). METHODS: Osteochondral autografting was developed and optimized in canine cadavers and purpose-bred research dogs using the Osteochondral Autograft Transfer System (OATS). Dogs with stifle OC (n=10 dogs, 12 stifles) were then treated using the OATS system. Outcomes were assessed by radiography (n=12), magnetic resonance imaging (1), second-look arthroscopy (9), lameness scoring (12), and telephone survey of owners (10 clients, 12 stifles) 6-15 months after surgery. RESULTS: Complications were documented in 4 of the 12 stifles treated and included peri-incisional seromas (3) and marked stifle effusion (1). Subjective assessment of follow-up radiographs revealed evidence of integration of the grafts with maintenance of subchondral bone surface architecture. Subjective assessment of follow-up MRI in 1 stifle revealed evidence for incorporation of grafts with restoration of articular surface contour. Second-look arthroscopy 6-30 weeks after surgery revealed maintenance of articular cartilage at the graft site. Dogs were significantly (P<.001) less lame at follow-up compared with preoperative scores. Based on follow-up owner surveys, only 2 dogs had no pain or lameness; the other dogs were judged to have mild pain and/or lameness. All owners noticed improvement in the dogs' quality of life after surgery. CONCLUSION: Osteochondral autografting deserves consideration and further evaluation as a primary treatment option for stifle OC in dogs. CLINICAL RELEVANCE: Osteochondral autografting for treatment of lateral femoral condylar OC lesions in dogs using OATS instrumentation is safe and results in improved function and quality of life based on owners' perception 6-15 months after treatment.

Early lesions of osteochondrosis in the distal tibia of foals.

Material available for research into osteochondrosis (OC) in humans tends to represent chronic lesions. Comparative studies of early lesions in young animals are, therefore, important in clarifying the pathogenesis of OC in humans. Recent studies in pigs provide strong evidence that lesions of articular OC are associated with a focal failure in the cartilage canal vascular supply to epiphyseal growth cartilage (articular-epiphyseal cartilage complex excluding the articular cartilage). The purpose of the present study was to examine histological sections from a specific predilection site for articular OC in the distal tibia of a large number of young foals to determine if the same is true in horses. Material from the distal tibiae of 100 foals aged from 191 days of gestation to 153 days old was collected from routine submissions of fetuses and foals for post mortem examination. The tibiae were band-sawed into slabs, and selected slabs were processed for histology, stained with hematoxylin and eosin, and examined using light microscopy. Early subclinical developmental stages of OC were found in the most common site for clinical OC lesions of horses in nine of 100 foals aged 12 to 122 days old. All lesions contained areas of chondrocyte necrosis that were associated with cartilage canal necrosis in five of nine foals. Five of these foals also had focal disruption of enchondral ossification at the chondro-osseous junction in the same site. Early lesions purported to play a role in the initial stages of articular OC in the distal tibia of horses were characterized by chondrocyte necrosis and likely occurred secondary to a failure of cartilage canal vascular supply to epiphyseal growth cartilage. The similarities in appearance between early lesions of piglets and foals suggest that information gained in one species may be transferable to others, including humans.

Hypertrophy and physiological death of equine chondrocytes in vitro.

REASON FOR PERFORMING STUDY: Equine osteochondrosis results from a failure of endochondral ossification during skeletal growth. Endochondral ossification involves chondrocyte proliferation, hypertrophy and death. Until recently no culture system was available to study these processes in equine chondrocytes. OBJECTIVE: To optimise an in vitro model in which equine chondrocytes can be induced to undergo hypertrophy and physiological death as seen in vivo. METHODS: Chondrocytes isolated from fetal or older (neonatal, growing and mature) horses were cultured as pellets in 10% fetal calf serum (FCS) or 10% horse serum (HS). The pellets were examined by light and electron microscopy. Total RNA was extracted from the pellets, and quantitative PCR carried out to investigate changes in expression of a number of genes regulating endochondral ossification. RESULTS: Chondrocytes from fetal foals, grown as pellets, underwent hypertrophy and died by a process morphologically similar to that seen in vivo. Chondrocytes from horses age >5 months did not undergo hypertrophy in pellet culture. They formed intramembranous inclusion bodies and the cultures included cells of osteoblastic appearance. Pellets from neonatal foals cultured in FCS resembled pellets from older horses, however pellets grown in HS underwent hypertrophy but contained inclusion bodies. Chondrocytes from fetal foals formed a typical cartilage-like tissue grossly and histologically, and expressed the cartilage markers collagen type II and aggrecan mRNA. Expression of Sox9, collagen type II, Runx2, matrix metalloproteinase-13 and connective tissue growth factor mRNA increased at different times in culture. Expression of fibroblast growth factor receptor-3 and vascular endothelial growth factor mRNA decreased with time in culture. CONCLUSIONS: Freshly isolated cells from fetal growth cartilage cultured as pellets provide optimal conditions for studying hypertrophy and death of equine chondrocytes. POTENTIAL RELEVANCE: This culture system should greatly assist laboratory studies aimed at elucidating the pathogenesis of osteochondrosis.

Osteochondrosis and copper: histology of articular cartilage from foals out of copper supplemented and non-supplemented dams.

Copper (Cu) supplementation of dams in late gestation may be protective against articular cartilage abnormalities in foals. Articular cartilage was harvested from 22 Thoroughbred foals at 160 days of age, at sites predisposed to osteochondrosis (OC), and examined for evidence of early cartilage abnormalities and established dyschondroplastic (DCP) lesions to determine if there were any significant differences due to mare Cu supplementation by injection during late gestation, or foal liver Cu concentration. Cu supplemented mares received calcium Cu edetate injections in late gestation (250 mg at around 220, 248, 276 and 304 days gestation, then every two weeks until foaling). Foals were euthanased at 160 days of age and articular cartilage was harvested from four defined sites. Samples were examined for histological appearance of chondrocytes after staining with haematoxylin and eosin, and were also stained with toluidine blue to indicate proteoglycan content. Alkaline phosphatase (ALP) activity was detected by histochemistry, and histocytochemical techniques were used to determine the expression of cathepsin B. Cu supplementation of the dam, or liver Cu concentration of the foal at birth or 160 days of age had no statistically significant effect on the frequency of cartilage irregularities observed grossly, or abnormalities detected histologically at four defined sites. ALP expression was similar in all samples. Cathepsin B expression varied between sites, and was seen in chondrocyte clusters. The intensity of toludine blue staining varied between sites. Minor histological cartilage abnormalities were observed in cartilage from clinically normal animals. These abnormalities might be 'early' dyschondroplastic lesions, which could resolve or progress. The role of Cu in the development, resolution or progression of dyschondroplastic lesions is poorly understood.

Effects of atelocollagen gel containing bone marrow-derived stromal cells on repair of osteochondral defect in a dog.

To clarify the contribution of autologous transplantation of mesenchymal stromal cells (MSCs), an atelocollagen gel containing or not containing fluorescently-labeled canine MSCs was transplanted into an osteochondral defect which did not repair spontaneously and the histological repair of the defect was compared. Although an early repair of the cartilage was not observed in either defect, the reproduction of subchondral bone was remarkable in the MSCs-implanted defect. Moreover, in 2 weeks after operation, the implanted MSCs were located in the deeper regions of the defect, suggesting the differentiation of osteoblasts. There was a possibility that the movement of the implanted MSCs was due to an increase in intra-articular pressure from postoperative inflammation.

Identification of infrared absorption spectral characteristics of synovial fluid of horses with osteochondrosis of the tarsocrural joint.

OBJECTIVE: To determine the feasibility of the use of Fourier-transform infrared (FTIR) spectroscopy within the midinfrared range to differentiate synovial fluid samples of joints with osteochondrosis from those of control samples. ANIMALS: 33 horses with osteochondrosis of the tarsocrural joint and 31 horses free of tarsocrural joint disease. PROCEDURES: FTIR spectroscopy of synovial fluid was used. Sixty-four synovial fluid samples from the tarsocrural joint were collected. Of these, 33 samples were from horses with radiographic evidence of osteochondrosis of the tarsocrural joint and 31 from control joints. Disease-associated features within infrared spectra of synovial fluid were statistically selected for spectral classification, and the variables identified were used in a classification model. Linear discriminant analysis and leave-one-out cross-validation were used to develop a classifier to identify joints with osteochondrosis. RESULTS: 12 significant subregions were identified that met the selection criteria. The stepwise discriminant procedure resulted in the final selection of 6 optimal regions that most contributed to the discriminatory power of the classification algorithm. Infrared spectra derived from synovial fluid of joints with osteochondrosis were differentiated from the control samples with accuracy of 77% (81% specificity and 73% sensitivity). CONCLUSIONS AND CLINICAL RELEVANCE: The disease-associated characteristics of infrared spectra of synovial fluid from joints with osteochondrosis may be exploited via appropriate feature selection and classification algorithms to differentiate joints with osteochondrosis from those of control joints. Further study with larger sample size including age-, breed-, and sex-matched control horses would further validate the clinical value of infrared spectroscopy for the diagnosis of osteochondrosis in horses.

Evaluation of the influences of exercise, birth date, and osteochondrosis on plasma bone marker concentrations in Hanoverian Warmblood foals.

OBJECTIVE: To determine whether plasma concentrations of bone turnover markers in growing Hanoverian foals are influenced by age, housing conditions, or osteochondrosis. ANIMALS: 165 healthy foals and 119 foals with osteochondrosis. PROCEDURES: Foals were allocated according to birth date and housing management into groups of early-born (born before March 31, 2001; n = 154 foals, 88 of which were healthy and 66 of which had osteochondrosis) and late-born (born after March 31, 2001; 130 foals, 77 of which were healthy and 53 of which had osteochondrosis) foals. Plasma osteocalcin and carboxyterminal propeptide of type I collagen concentrations were analyzed as markers of bone formation, and carboxyterminal telopeptide of type I collagen concentration was analyzed as a marker of bone resorption. Foals underwent radiographic evaluation to screen for osteochondrosis. RESULTS: Plasma concentrations of osteocalcin, carboxyterminal propeptide of type I collagen, and carboxyterminal telopeptide of type I collagen decreased with age, but these changes were more distinct in late-born foals than in early-born foals. Neither sex nor predisposition to develop osteochondrosis affected the pattern of bone marker changes in either group. CONCLUSIONS AND CLINICAL RELEVANCE: An age-related decrease in concentrations of bone markers was seen during the first 200 days of life. Changes in bone marker concentrations were similar for foals with osteochondrosis and healthy foals. The correlation between the decrease in bone marker concentration and date of birth indicates that there are differences in skeletal development between early- and late-born foals.

Osteochondral fragments involving the dorsomedial aspect of the proximal interphalangeal joint in young horses: 6 cases (1997-2006).

OBJECTIVE: To determine clinical and diagnostic imaging findings in young horses with osteochondral fragments involving the dorsomedial aspect of the proximal interphalangeal (PIP) joint. DESIGN: Retrospective case series. ANIMALS: 6 horses. PROCEDURES: Medical records were reviewed. Follow-up information was obtained through telephone conversations with owners or trainers or by examining race records. RESULTS: Horses were between 1 and 4 years old. Three had bilateral osteochondral fragments in the forelimbs (n = 2 horses) or hind limbs (1). Radiographically, all but 1 fragment seemed to originate from the dorsomedial aspect of the distal end of the first phalanx. Fragment size ranged from 6 x 9 mm to 11 x 21 mm. Three horses had lameness referable to the region of the affected joint; the other 3 horses did not have clinical signs referable to affected PIP joints. Two horses were euthanized shortly after diagnosis at the owners' request because of concerns that the horses would be unsuited for their intended athletic use. Two of the 3 horses in which fragments were incidental findings were able to race successfully, although 1 received intra-articular corticosteroid treatments; the third was retired because of unrelated orthopedic problems. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that osteochondral fragments involving the dorsomedial aspect of the PIP joint may be an incidental finding in young horses. Given the absence of clinical signs in 5 of 9 affected joints and the fact that 3 of 6 horses were affected bilaterally, a developmental origin of the fragments was suspected.

[Prevalence of hereditary diseases in three-year-old Swiss Warmblood horses]. / Erhebung der Prävalenz von Erbkrankheiten bei dreijährigen Schweizer Warmblutpferden.

The objective of this study was to investigate clinical signs indicating hereditary diseases like equine sarcoid, osteochondrosis (OC) and the idiopathic laryngeal hemiplegia (ILH), and to demonstrate relationships between environment, feeding habits and conformation ("exterieur" evaluation) of the horses. For this purpose, we analyzed veterinary examinations of 403 stallions at the approvals since 1994 examined 493 three-year-old Swiss Warmblood horses, which were shown at the Swiss-Field-Tests in 2005. With the help of the owners a questionnaire on health, environment and feeding habits of the animals was completed. At the same time, the horses were assessed and graded for their "exterieur" (type, conformation, gaits) by judges of the Swiss Sporthorse breeding association. In 11.5% of horses sarcoids were found, 8.7% showed one and 2.8% several tumors. The prevalence of sarcoids in offspring of sires with known sarcoids was not significantly higher than in descendants from stallions without a known history of sarcoids. We found distended joints as a possible symptom of OC in 11.4% of the horses, 3.9% (n = 19) in both tarsal joints. We did not find a relationship between enlarged joints in the offspring and the presence of OC in the sires. Abnormal respiratory noise at work, as a possible sign for ILH, was heard only in 1.2% (n = 6). It is important to note that while we found a high number of sarcoid affected horses compared to other studies, presence of enlarged joints was not very frequent and very few horses showed abnormal respiratory noise. Additionally, we found no correlation between "exterieur" marks and the horse's general health.