In articular cartilage (AC), the collagen arcades provide the tissue with its extraordinary mecha... more In articular cartilage (AC), the collagen arcades provide the tissue with its extraordinary mechanical properties. As these structures cannot be restored once damaged, functional restoration of AC defects remains a major challenge. We report that the use of a converged bioprinted, osteochondral implant, based on a gelatin methacryloyl cartilage phase, reinforced with precisely patterned melt electrowritten polycaprolactone micrometer‐scale fibers in a zonal fashion, inspired by native collagen architecture, can provide long‐term mechanically stable neo‐tissue in an orthotopic large animal model. The design of this novel implant was achieved via state‐of‐the‐art converging of extrusion‐based ceramic printing, melt electrowriting, and extrusion‐based bioprinting. Interestingly, the cell‐free implants, used as a control in this study, showed abundant cell ingrowth and similar favorable results as the cell‐containing implants. Our findings underscore the hypothesis that mechanical stabi...
Objective: We investigate the potential of a prototype multimodality arthroscope, combining ultra... more Objective: We investigate the potential of a prototype multimodality arthroscope, combining ultrasound, optical coherence tomography (OCT) and arthroscopic indentation device, for assessing cartilage lesions, and compare the reliability of this approach with conventional arthroscopic scoring ex vivo. Design: Areas of interest (AIs, N ¼ 43) were selected from equine fetlock joints (N ¼ 5). Blind-coded AIs were independently scored by two equine surgeons employing International Cartilage Repair Society (ICRS) scoring system via conventional arthroscope and multimodality arthroscope, in which highfrequency ultrasound and OCT catheters were attached to an arthroscopic indentation device. In addition, cartilage stiffness was measured with the indentation device, and lesions in OCT images scored using custom-made automated software. Measurements and scorings were performed twice in two separate rounds. Finally, the scores were compared to histological ICRS scores. Results: OCT and arthroscopic examinations showed the highest average agreements (55.2%) between the scoring by surgeons and histology scores, whereas ultrasound had the lowest (50.6%). Average intraobserver agreements of surgeons and interobserver agreements between rounds were, respectively, for conventional arthroscope (68.6%, 69.8%), ultrasound (68.6%, 68.6%), OCT (65.1%, 61.7%) and automated software (65.1%, 59.3%). Conclusions: OCT imaging supplemented with the automated software provided the most reliable lesion scoring. However, limited penetration depth of light limits the clinical potential of OCT in assessing human cartilage thickness; thus, the combination of OCT and ultrasound could be optimal for reliable diagnostics. Present findings suggest imaging and quantitatively analyzing the entire articular surface to eliminate surgeon-related variation in the selection of the most severe lesion to be scored.
Conventional arthroscopic evaluation of articular cartilage is subjective and poorly reproducible... more Conventional arthroscopic evaluation of articular cartilage is subjective and poorly reproducible. Therefore, implementation of quantitative diagnostic techniques, such as near infrared spectroscopy (NIRS) and optical coherence tomography (OCT), is essential. Locations (n = 44) with various cartilage conditions were selected from mature equine fetlock joints (n = 5). These locations and their surroundings were measured with NIRS and OCT (n = 530). As a reference, cartilage proteoglycan (PG) and collagen contents, and collagen network organization were determined using quantitative microscopy. Additionally, lesion severity visualized in OCT images was graded with an automatic algorithm according to International Cartilage Research Society (ICRS) scoring system. Artificial neural network with variable selection was then employed to predict cartilage composition in the superficial and deep zones from NIRS data, and the performance of two models, generalized (including all samples) and condition-specific models (based on ICRS-grades), was compared. Spectral data correlated significantly (p < 0.002) with PG and collagen contents, and collagen orientation in the superficial and deep zones. The combination of NIRS and OCT provided the most reliable outcome, with condition-specific models having lower prediction errors (9.2%) compared to generalized models (10.4%). Therefore, the results highlight the potential of combining both modalities for comprehensive evaluation of cartilage during arthroscopy. Articular cartilage is an aneural tissue that enables near frictionless contact between articulating bones. The tissue consists mainly of water, collagen and proteoglycans (PGs) 1 , and due to its limited regenerative capability, it is susceptible to progressive degeneration after trauma and wear. Cartilage defects can result from various reasons and lead to degenerative conditions, such as osteoarthritis (OA) and trauma-induced post-traumatic OA (PTOA), affecting millions of people worldwide 2, 3 . Although these diseases have similar initial stages, including superficial PG loss and disruption of superficial collagen network, PTOA mainly impacts the young, whereas OA mostly the elderly. These alterations in cartilage composition and structure often lead to excessive strains within the tissue, and therefore progressive deterioration. As no cure currently exists for OA, it would be essential to reliably diagnose cartilage injuries and evaluate their surrounding tissue to prevent the initiation and progression of PTOA, e.g. via surgical intervention. Arthroscopic repair surgery is often performed to treat joint conditions diagnosed via clinical examination, computed tomography (CT) or magnetic resonance imaging (MRI). Due to poor soft tissue contrast in conventional CT images and relatively poor resolution of clinical MRI 4 , arthroscopic examination may reveal previously
The equine fetlock joint has the largest number of traumatic and degenerative lesions of all join... more The equine fetlock joint has the largest number of traumatic and degenerative lesions of all joints of the appendicular skeleton. To gain insight into the distribution of cartilage degeneration across the articular surface in relation to age in order better to understand the dynamic nature and progression of osteoarthritis (OA). That there would be a specific age-related distribution pattern of cartilage degeneration in the equine metacarpophalangeal joint. The proximal articular cartilage surfaces of the first phalanges (P1) of 73 slaughter horses (age range 0.4-23 years) with different stages of osteoarthritis were scored semiquantitatively on a 0 to 5 scale and also assessed quantitatively using the cartilage degeneration index (CDI(P1)), which ranges from 0 to 100%. Furthermore, CDI values were determined for special areas of interest; medial dorsal surface (CDI(mds)), lateral dorsal surface (CDI(lds)), medial central fovea (CDI(mcf)) and lateral central fovea (CDI(lcf)). Correlations were calculated for CDI(P1) values and CDI values at the specific areas of interest with macroscopic scores and with age. There was a high correlation between the semiquantitative macroscopic score and the quantitative CDI(P1) values (r = 0.92; P &lt; 0.001). A macroscopic score of 0 (i.e. no obvious cartilage degeneration) corresponded with a CDI(P1) mean +/- s.e. value of 25 +/- 2.8% and a macroscopic score of 5 (i.e. severe cartilage degeneration in localised areas) with a mean +/- s.e. value of 38.1 +/- 7.9%. There was a moderate but highly significant correlation between the CDI(P1) value and the age of the horses (r = 0.41; P &lt; 0.001). Highest CDI values were calculated for the medial dorsal surface (from 10.6 +/- 2.8% at macroscopic Grade 0 to 63.1 +/- 8.4% at Grade 5). At the lateral dorsal surface, these values were 5.9 +/- 1.4% and 47.2 +/- 10.4%, respectively. The CDI(mcf) and CDI(lcf) were significantly lower (P &lt; 0.05) than the CDI(mds) and CDI(lds) at all grades. The CDI(mcf) ranged from 1.0 +/- 2.9% at Grade 0 to 43.7 +/- 9.1% at Grade 5; laterally, these values were 1.5 +/- 2.6% and 15.2 +/- 6.2%, respectively. CDI grading increased from lateral to medial and from central to dorsal. This specific distribution pattern confirms the heterogeneous nature of the OA process and strongly supports an important role for biomechanical loading, superimposed on age-related changes, in the spread of the disorder over the joint. Knowledge of the development of OA across the articular surface is essential for understanding the dynamic nature and progression of the disease and can form a basis for improvements in diagnostic and therapeutic approaches to degenerative joint disease.
Reasons for performing study: The concept of functional adapatation of articular cartilage during... more Reasons for performing study: The concept of functional adapatation of articular cartilage during maturation has emerged from earlier biochemical research. However, articular cartilage has principally a biomechanical function governed by joint loading. Objectives: To verify whether the concept of functional adaptation can be confirmed by direct measurement of biomechanical properties of cartilage. Hypothesis: Fetuses have homogeneous (i.e. site-independent) cartilage with regard to biomechanical properties. During growth and development to maturity, the biomechanical characteristics adapt according to functional (loading) demands, leading to distinct, site-dependent biomechanical heterogeneity of articular cartilage. Methods: Osteochondral plugs were drilled out of the surface at 2 differently loaded sites (Site 1: intermittent impactloading during locomotion, Site 2: low-level constant loading during weightbearing) of the proximal articular cartilage surface of the proximal phalanx in the forelimb from stillborn foals (n = 8), horses of age 5 (n = 9) and 18 months (n = 9) and mature horses (n = 13). Cartilage thickness was measured using ultrasonic, optical and needle-probe techniques. The osteochondral samples were biomechanically tested in indentation geometry. Young's modulus at equilibrium, dynamic modulus at 1 Hz and the ratios of these moduli values between Sites 1 and 2 were calculated. Age and site effects were evaluated statistically using ANOVA tests. The level of significance was set at P<0.05. Results: Fetal cartilage was significantly thicker compared to the other ages with no further age-dependent differences in cartilage thickness from age 5 months onwards. Young's modulus stayed constant at Site 1, whereas at Site 2 there was a gradual, statistically significant increase in modulus during maturation. Values of dynamic modulus at both Sites 1 and 2 were significantly higher in the fetus and decreased after birth. Values for both moduli were significantly different between Sites 1 and 2 from age 18 months onwards. The ratio of values between Sites 1 and 2 for Young's modulus and dynamic modulus showed a gradual decrease from ~1.0 at birth to 0.5-0.6 in the mature horse. At age 18 months, all values were comparable to those in the mature horse. Conclusions: In line with the concept of functional adaptation, the neonate is born with biomechanically 'blank' or homogeneous cartilage. Functional adaptation of biomechanical properties takes place early in life, resulting in cartilage with a distinct heterogeneity in functional characteristics. At age 18 months, functional adaptation, as assessed by the biomechanical characteristics, has progressed to a level comparable to the mature horse and, after this age, no major adaptations seem to occur. Potential relevance: Throughout life, different areas of articular cartilage are subjected to different types of loading. Differences in loading can adequately be met only when the tissue is biomechanically adapted to withstand these different loading conditions without injury. This process of functional adaptation starts immediately after birth and is completed well before maturity. This makes the factor of loading at a young age a crucial variable, and emphasises the necessity to optimise joint loading during early life in order to create an optimal biomechanical quality of articular cartilage, which may well turn out to be the best prevention for joint injury later in life.
The case reported here describes an atypical presentation of cellulitis/lymphangitis in an 8-year... more The case reported here describes an atypical presentation of cellulitis/lymphangitis in an 8-year-old Dutch Warmblood mare. The horse was presented with a history of recurrent episodes of cellulitis/lymphangitis and the presence of fluctuating cyst-like lesions on the left hindlimb. These lesions appeared to be interconnected lymphangiectasias. Surgical debridement followed by primary wound closure and local drainage was performed under general anaesthesia. Twelve months post surgery, no recurrence of cellulitis/lymphangitis had occurred and the mare had returned to her former use as a dressage horse. bs_bs_banner
ABSTRACT This report describes an unusual case of unilateral abdominal cryptorchidism in a Shetla... more ABSTRACT This report describes an unusual case of unilateral abdominal cryptorchidism in a Shetland pony. On the cryptorchid side, no testicular structures were present in the inguinal area but testicular/epididymal remnants at the distal end of a pedunculated structure could be discerned in the abdomen at laparoscopic examination. Furthermore, the deferent duct was disconnected from the epididymal remnants and the deferent duct was visible at the caudal aspect of the bladder, traversing over the lateral ligament of the bladder into the vaginal ring where it ended blindly in the vaginal process. After laparoscopic removal of the testicular/epididymal remnants and castration of the contralateral normally descended testicle, the hCG stimulation test was negative and the pony no longer exhibited masculine behaviour.
The importance of the equine thoracolumbar vertebral column in orthopaedic disorders is well reco... more The importance of the equine thoracolumbar vertebral column in orthopaedic disorders is well recognized and diagnostic imaging becomes more feasible, but little is known about variations in the anatomical configuration within breeds. In this descriptive post-mortem study, anatomical variations in three widely differing breeds: Warmblood horses, Shetland ponies and semi-feral Konik horses are described. The caudal cervical (C), thoracic (T), lumbar (L) and sacral (S) regions of the vertebral column of 30 Warmblood horses, 29 Shetland ponies and 18 Konik horses were examined using computed tomography and visualized by volume rendering. Homologous/ morphologic variations in the caudal cervical area were frequently seen in Warmblood horses (43%), which was significantly more than in the other breeds (p < 0.001). The as standard described equine formula of 18 T, 6 L and 5 S vertebrae was seen in 78% of Konik horses, but only in 53% Warmblood horses and 38% Shetland ponies, which was significantly different (p < 0.05). Overall, Shetland ponies showed a higher tendency of thoracoization, lumbarization and more variations in the number of vertebrae and pairs of ribs. Ankylosed intertransverse joints (ITJs) between transverse processes of the lumbar vertebrae were most common between the second last and last lumbar vertebra and prevalence was significantly higher in Shetland ponies (61%), than in Warmblood horses (38%) and Konik horses (7%) (p < 0.0001). Cranial to the second last lumbar vertebra there were fewer ITJs ankylosed (14%) in Warmblood horses (p < 0.0095), and this decrease in number of ankylosed ITJs was different compared to the change in ankylosed ITJs in Shetland ponies (p < 0.005). ITJs occurred asymmetrically in 15% (12/77) of the cases. A limitation of the study was that clinical data of the horses were only incompletely available, precluding any conclusions about the potential clinical implications of anatomical variations. Knowledge of variation in osseous anatomy of the equine thoracolumbar vertebral column is important for the interpretation of diagnostic imaging. To assess the functional importance and clinical relevance of this variation, follow-up studies are necessary.
Objective-To evaluate an arthroscopic indentation instrument (Artscan 200) for assessment of the ... more Objective-To evaluate an arthroscopic indentation instrument (Artscan 200) for assessment of the health status of equine articular cartilage. Study Design-In vitro experiment using equine isolated proximal phalanx (P1) specimens. Sample Population-P1 specimens from 39 horses (aged 1.5-22 years). Methods-Reproducibility was tested by determination of the coefficient of variation (CV). Dynamic modulus and cartilage degeneration index (CDI) values were measured at 2 predefined sites (site 1, joint margin; site 2, joint center) to assess the accuracy and to evaluate the relation with surface integrity. Results-CV was 9.0%. A significant decrease in indenter force was identified when dynamic modulus values decreased to o2.5 MPa (range of tested samples 0.9-8.1 MPa) and when CDI values at site 1 increased to 450% (range 5.4-72.8%). Conclusions-Technique reproducibility was adequate but accuracy was limited. The device identified degeneration-associated decreases in cartilage stiffness only when the mechanical properties of the cartilage were considerably changed. Clinical Relevance-Usefulness of this indentation instrument during arthroscopic surgery would be limited in the initial phase of OA-like cartilage degeneration, but may yield important information in more advanced OA.
for performing study: Osteoarthritis (OA) is one of the most prevalent and disabling chronic cond... more for performing study: Osteoarthritis (OA) is one of the most prevalent and disabling chronic conditions affecting horses and leads to degeneration of articular cartilage. Diagnosis is based on clinical signs in combination with radiography, which is relatively insensitive and provides only an indication of accumulated damage. Alternative methods, such as molecular markers, are therefore needed that can quantitatively, reliably and sensitively detect osteoarthritic changes in the joints at an early stage of the disease. If such markers are to be used reliably, it is important to know the relationship between marker concentration and cartilage composition. Objectives: To study the relationship between cartilage composition, synovial fluid levels of glycosaminoglycans (GAGs), hydroxyproline (Hyp) and general matrix metalloproteinase (MMP) activity, and the presence and severity of articular cartilage damage on the articular surface of P1. Methods: Synovial fluid (SF) was collected from the metacarpophalangeal joints of 60 mature horses, and levels of GAGs, Hyp and general MMP activity were determined. Further, GAG and denatured collagen content of the articular cartilage were determined at the dorsal articular margin of P1 (site 1) and central cavity (site 2). The presence and severity of cartilage change was quantified using the cartilage degeneration index (CDI), measured at the same 2 sites. Correlations between SF parameters, cartilage composition and degree of cartilage degeneration were sought using correlation analysis. Results: There was no correlation between GAG or Hyp content of SF and the amount of GAGs or denatured collagen, respectively, in cartilage. In joints with moderate to severe cartilage damage, the GAG content of site 1 was significantly lower than in joints with no to minimal cartilage change (P = 0.005) and there was a negative correlation between the amount of denatured collagen and GAG content at site 1 in all joints (r = -0.39, P = 0.002). Further, in joints with moderate to severe cartilage damage, there was a significant positive correlation between MMP activity in SF and Hyp levels in SF (r = 0.72, P<0.001) and CDI at sites 1 (r = 0.46, P=0.03) and 2 (r = 0.43, P = 0.04). Conclusions: General MMP activity in joints with moderate to severe cartilage damage is related to the severity of those cartilage changes and to Hyp levels in SF. Glycosaminoglycan levels in SF are not directly related to MMP activity, GAG content of articular cartilage or severity of cartilage change. Potential relevance: Glycosaminoglycan levels in SF are not helpful for the early detection of cartilage lesions. In damaged joints, Hyp levels may give an indication of the severity of cartilage change as they are strongly related to MMP activity, but do not qualify as markers for the presence or absence of cartilage lesions.
for performing study: Training at a very young age may influence the characteristics of the colla... more for performing study: Training at a very young age may influence the characteristics of the collagen network of articular cartilage extracellular matrix (ECM) in horses. Objectives: To investigate whether increasing workload of foals results in significant changes in the biochemical composition of articular cartilage ECM. Methods: Thoroughbred foals (n = 33) were divided into 2 different exercise groups from age 10 days-18 months. One group (PASTEX; n = 15) was reared at pasture; the other (CONDEX; n = 18) underwent a specific additional training programme that increased workload by 30%. At mean age 18 months, 6 animals from each group were subjected to euthanasia. The proximal articular surface of the proximal phalanx of the right hindlimb was examined for the presence of damage using the cartilage degeneration index (CDI). Samples were taken from 2 sites with known different loading patterns. Slices were analysed for DNA, glycosaminoglycans (GAG), collagen and post translational modifications of collagen (formation of hydroxylysylpyridinoline [HP] and pentosidine crosslinks, and hydroxylysine [Hyl]), and exercise groups and different sites compared. Results: There were no differences in CDI between PASTEX and CONDEX animals, indicating the absence of extra joint damage due to the exercise regimen. There were site-related differences for most biochemical variables, corroborating earlier reports. All biochemical variables showed differences between PASTEX and CONDEX groups at one of the sites, and some at both. GAG and collagen levels were lower in the CONDEX group whereas Hyl, HP crosslinks and pentosidine crosslinks were higher. Conclusions and potential relevance: A measurable effect of the conditioning exercise was demonstrated. The margin between too much and too little work when training foals may be narrower than intuitively presumed.
Records of lame horses (2004-2007) with (peri-)synovial pain of the metacarpophalangeal joint (MC... more Records of lame horses (2004-2007) with (peri-)synovial pain of the metacarpophalangeal joint (MCPJ, n = 53), digital flexor tendon sheath (DFTS, n = 20), and distal interphalangeal joint (DIPJ, n = 31) were evaluated (n = 104). Cases were grouped according to the following treatments: (1) systemic administration of NSAIDs (n = 40); (2) intrasynovial administration of glucocorticosteroids (n = 30); (3) combination treatment (n = 34). Outcome was defined as successful when the owner reported that the horse had returned to its previous level of performance within a period of up to six months after treatment. Statistical evaluation was performed using Fisher&#39;s Exact tests and multivariable logistic regression analyses at P &lt; 0.05. Successful outcome was significantly higher for horses in treatment group 3 (19/34, 55.9 per cent, P = 0.021) than in treatment groups 1 (11/40, 27.5 per cent, OR 0.21, 95 per cent CI 0.06 to 0.71) and 2 (8/30, 26.6 per cent, OR 0.18, 95 per cent CI 0.06 to 0.56). A significant association was only found with regard to the affected synovial structure (P = 0.025); relative to the DIPJ, involvement of the MCPJ and DFTS was four to five times more likely to provide a successful outcome (OR 4.18 and 5.59, 95 per cent CI 1.24 to 14.08 and 1.42-22.22, respectively).
Arthroscopy enables direct inspection of the articular surface, but provides no information on de... more Arthroscopy enables direct inspection of the articular surface, but provides no information on deeper cartilage layers. Optical coherence tomography (OCT), based on measurement of reflection and backscattering of light, is a diagnostic technique used in cardiovascular surgery and ophthalmology. It provides cross-sectional images at resolutions comparable to that of low-power microscopy. The aim of this study was to determine if OCT is feasible for advanced clinical assessment of lesions in equine articular cartilage during diagnostic arthroscopy. Diagnostic arthroscopy of 36 metacarpophalangeal joints was carried out ex vivo. Of these, 18 joints with varying degrees of cartilage damage were selected, wherein OCT arthroscopy was conducted using an OCT catheter (diameter 0.9 mm) inserted through standard instrument portals. Five sites of interest, occasionally supplemented with other locations where defects were encountered, were arthroscopically graded according to the International Cartilage Repair Society (ICRS) classification system. The same sites were evaluated qualitatively (ICRS classification and morphological description of the lesions) and quantitatively (measurement of cartilage thickness) on OCT images. OCT provided high resolution images of cartilage enabling determination of cartilage thickness. Comparing ICRS grades determined by both arthroscopy and OCT revealed poor agreement. Furthermore, OCT visualised a spectrum of lesions, including cavitation, fibrillation, superficial and deep clefts, erosion, ulceration and fragmentation. In addition, with OCT the arthroscopically inaccessible area between the dorsal MC3 and P1 was reachable in some cases. Arthroscopically-guided OCT provided more detailed and quantitative information on the morphology of articular cartilage lesions than conventional arthroscopy. OCT could therefore improve the diagnostic value of arthroscopy in equine orthopaedic surgery.
In articular cartilage (AC), the collagen arcades provide the tissue with its extraordinary mecha... more In articular cartilage (AC), the collagen arcades provide the tissue with its extraordinary mechanical properties. As these structures cannot be restored once damaged, functional restoration of AC defects remains a major challenge. We report that the use of a converged bioprinted, osteochondral implant, based on a gelatin methacryloyl cartilage phase, reinforced with precisely patterned melt electrowritten polycaprolactone micrometer‐scale fibers in a zonal fashion, inspired by native collagen architecture, can provide long‐term mechanically stable neo‐tissue in an orthotopic large animal model. The design of this novel implant was achieved via state‐of‐the‐art converging of extrusion‐based ceramic printing, melt electrowriting, and extrusion‐based bioprinting. Interestingly, the cell‐free implants, used as a control in this study, showed abundant cell ingrowth and similar favorable results as the cell‐containing implants. Our findings underscore the hypothesis that mechanical stabi...
Objective: We investigate the potential of a prototype multimodality arthroscope, combining ultra... more Objective: We investigate the potential of a prototype multimodality arthroscope, combining ultrasound, optical coherence tomography (OCT) and arthroscopic indentation device, for assessing cartilage lesions, and compare the reliability of this approach with conventional arthroscopic scoring ex vivo. Design: Areas of interest (AIs, N ¼ 43) were selected from equine fetlock joints (N ¼ 5). Blind-coded AIs were independently scored by two equine surgeons employing International Cartilage Repair Society (ICRS) scoring system via conventional arthroscope and multimodality arthroscope, in which highfrequency ultrasound and OCT catheters were attached to an arthroscopic indentation device. In addition, cartilage stiffness was measured with the indentation device, and lesions in OCT images scored using custom-made automated software. Measurements and scorings were performed twice in two separate rounds. Finally, the scores were compared to histological ICRS scores. Results: OCT and arthroscopic examinations showed the highest average agreements (55.2%) between the scoring by surgeons and histology scores, whereas ultrasound had the lowest (50.6%). Average intraobserver agreements of surgeons and interobserver agreements between rounds were, respectively, for conventional arthroscope (68.6%, 69.8%), ultrasound (68.6%, 68.6%), OCT (65.1%, 61.7%) and automated software (65.1%, 59.3%). Conclusions: OCT imaging supplemented with the automated software provided the most reliable lesion scoring. However, limited penetration depth of light limits the clinical potential of OCT in assessing human cartilage thickness; thus, the combination of OCT and ultrasound could be optimal for reliable diagnostics. Present findings suggest imaging and quantitatively analyzing the entire articular surface to eliminate surgeon-related variation in the selection of the most severe lesion to be scored.
Conventional arthroscopic evaluation of articular cartilage is subjective and poorly reproducible... more Conventional arthroscopic evaluation of articular cartilage is subjective and poorly reproducible. Therefore, implementation of quantitative diagnostic techniques, such as near infrared spectroscopy (NIRS) and optical coherence tomography (OCT), is essential. Locations (n = 44) with various cartilage conditions were selected from mature equine fetlock joints (n = 5). These locations and their surroundings were measured with NIRS and OCT (n = 530). As a reference, cartilage proteoglycan (PG) and collagen contents, and collagen network organization were determined using quantitative microscopy. Additionally, lesion severity visualized in OCT images was graded with an automatic algorithm according to International Cartilage Research Society (ICRS) scoring system. Artificial neural network with variable selection was then employed to predict cartilage composition in the superficial and deep zones from NIRS data, and the performance of two models, generalized (including all samples) and condition-specific models (based on ICRS-grades), was compared. Spectral data correlated significantly (p < 0.002) with PG and collagen contents, and collagen orientation in the superficial and deep zones. The combination of NIRS and OCT provided the most reliable outcome, with condition-specific models having lower prediction errors (9.2%) compared to generalized models (10.4%). Therefore, the results highlight the potential of combining both modalities for comprehensive evaluation of cartilage during arthroscopy. Articular cartilage is an aneural tissue that enables near frictionless contact between articulating bones. The tissue consists mainly of water, collagen and proteoglycans (PGs) 1 , and due to its limited regenerative capability, it is susceptible to progressive degeneration after trauma and wear. Cartilage defects can result from various reasons and lead to degenerative conditions, such as osteoarthritis (OA) and trauma-induced post-traumatic OA (PTOA), affecting millions of people worldwide 2, 3 . Although these diseases have similar initial stages, including superficial PG loss and disruption of superficial collagen network, PTOA mainly impacts the young, whereas OA mostly the elderly. These alterations in cartilage composition and structure often lead to excessive strains within the tissue, and therefore progressive deterioration. As no cure currently exists for OA, it would be essential to reliably diagnose cartilage injuries and evaluate their surrounding tissue to prevent the initiation and progression of PTOA, e.g. via surgical intervention. Arthroscopic repair surgery is often performed to treat joint conditions diagnosed via clinical examination, computed tomography (CT) or magnetic resonance imaging (MRI). Due to poor soft tissue contrast in conventional CT images and relatively poor resolution of clinical MRI 4 , arthroscopic examination may reveal previously
The equine fetlock joint has the largest number of traumatic and degenerative lesions of all join... more The equine fetlock joint has the largest number of traumatic and degenerative lesions of all joints of the appendicular skeleton. To gain insight into the distribution of cartilage degeneration across the articular surface in relation to age in order better to understand the dynamic nature and progression of osteoarthritis (OA). That there would be a specific age-related distribution pattern of cartilage degeneration in the equine metacarpophalangeal joint. The proximal articular cartilage surfaces of the first phalanges (P1) of 73 slaughter horses (age range 0.4-23 years) with different stages of osteoarthritis were scored semiquantitatively on a 0 to 5 scale and also assessed quantitatively using the cartilage degeneration index (CDI(P1)), which ranges from 0 to 100%. Furthermore, CDI values were determined for special areas of interest; medial dorsal surface (CDI(mds)), lateral dorsal surface (CDI(lds)), medial central fovea (CDI(mcf)) and lateral central fovea (CDI(lcf)). Correlations were calculated for CDI(P1) values and CDI values at the specific areas of interest with macroscopic scores and with age. There was a high correlation between the semiquantitative macroscopic score and the quantitative CDI(P1) values (r = 0.92; P &lt; 0.001). A macroscopic score of 0 (i.e. no obvious cartilage degeneration) corresponded with a CDI(P1) mean +/- s.e. value of 25 +/- 2.8% and a macroscopic score of 5 (i.e. severe cartilage degeneration in localised areas) with a mean +/- s.e. value of 38.1 +/- 7.9%. There was a moderate but highly significant correlation between the CDI(P1) value and the age of the horses (r = 0.41; P &lt; 0.001). Highest CDI values were calculated for the medial dorsal surface (from 10.6 +/- 2.8% at macroscopic Grade 0 to 63.1 +/- 8.4% at Grade 5). At the lateral dorsal surface, these values were 5.9 +/- 1.4% and 47.2 +/- 10.4%, respectively. The CDI(mcf) and CDI(lcf) were significantly lower (P &lt; 0.05) than the CDI(mds) and CDI(lds) at all grades. The CDI(mcf) ranged from 1.0 +/- 2.9% at Grade 0 to 43.7 +/- 9.1% at Grade 5; laterally, these values were 1.5 +/- 2.6% and 15.2 +/- 6.2%, respectively. CDI grading increased from lateral to medial and from central to dorsal. This specific distribution pattern confirms the heterogeneous nature of the OA process and strongly supports an important role for biomechanical loading, superimposed on age-related changes, in the spread of the disorder over the joint. Knowledge of the development of OA across the articular surface is essential for understanding the dynamic nature and progression of the disease and can form a basis for improvements in diagnostic and therapeutic approaches to degenerative joint disease.
Reasons for performing study: The concept of functional adapatation of articular cartilage during... more Reasons for performing study: The concept of functional adapatation of articular cartilage during maturation has emerged from earlier biochemical research. However, articular cartilage has principally a biomechanical function governed by joint loading. Objectives: To verify whether the concept of functional adaptation can be confirmed by direct measurement of biomechanical properties of cartilage. Hypothesis: Fetuses have homogeneous (i.e. site-independent) cartilage with regard to biomechanical properties. During growth and development to maturity, the biomechanical characteristics adapt according to functional (loading) demands, leading to distinct, site-dependent biomechanical heterogeneity of articular cartilage. Methods: Osteochondral plugs were drilled out of the surface at 2 differently loaded sites (Site 1: intermittent impactloading during locomotion, Site 2: low-level constant loading during weightbearing) of the proximal articular cartilage surface of the proximal phalanx in the forelimb from stillborn foals (n = 8), horses of age 5 (n = 9) and 18 months (n = 9) and mature horses (n = 13). Cartilage thickness was measured using ultrasonic, optical and needle-probe techniques. The osteochondral samples were biomechanically tested in indentation geometry. Young's modulus at equilibrium, dynamic modulus at 1 Hz and the ratios of these moduli values between Sites 1 and 2 were calculated. Age and site effects were evaluated statistically using ANOVA tests. The level of significance was set at P<0.05. Results: Fetal cartilage was significantly thicker compared to the other ages with no further age-dependent differences in cartilage thickness from age 5 months onwards. Young's modulus stayed constant at Site 1, whereas at Site 2 there was a gradual, statistically significant increase in modulus during maturation. Values of dynamic modulus at both Sites 1 and 2 were significantly higher in the fetus and decreased after birth. Values for both moduli were significantly different between Sites 1 and 2 from age 18 months onwards. The ratio of values between Sites 1 and 2 for Young's modulus and dynamic modulus showed a gradual decrease from ~1.0 at birth to 0.5-0.6 in the mature horse. At age 18 months, all values were comparable to those in the mature horse. Conclusions: In line with the concept of functional adaptation, the neonate is born with biomechanically 'blank' or homogeneous cartilage. Functional adaptation of biomechanical properties takes place early in life, resulting in cartilage with a distinct heterogeneity in functional characteristics. At age 18 months, functional adaptation, as assessed by the biomechanical characteristics, has progressed to a level comparable to the mature horse and, after this age, no major adaptations seem to occur. Potential relevance: Throughout life, different areas of articular cartilage are subjected to different types of loading. Differences in loading can adequately be met only when the tissue is biomechanically adapted to withstand these different loading conditions without injury. This process of functional adaptation starts immediately after birth and is completed well before maturity. This makes the factor of loading at a young age a crucial variable, and emphasises the necessity to optimise joint loading during early life in order to create an optimal biomechanical quality of articular cartilage, which may well turn out to be the best prevention for joint injury later in life.
The case reported here describes an atypical presentation of cellulitis/lymphangitis in an 8-year... more The case reported here describes an atypical presentation of cellulitis/lymphangitis in an 8-year-old Dutch Warmblood mare. The horse was presented with a history of recurrent episodes of cellulitis/lymphangitis and the presence of fluctuating cyst-like lesions on the left hindlimb. These lesions appeared to be interconnected lymphangiectasias. Surgical debridement followed by primary wound closure and local drainage was performed under general anaesthesia. Twelve months post surgery, no recurrence of cellulitis/lymphangitis had occurred and the mare had returned to her former use as a dressage horse. bs_bs_banner
ABSTRACT This report describes an unusual case of unilateral abdominal cryptorchidism in a Shetla... more ABSTRACT This report describes an unusual case of unilateral abdominal cryptorchidism in a Shetland pony. On the cryptorchid side, no testicular structures were present in the inguinal area but testicular/epididymal remnants at the distal end of a pedunculated structure could be discerned in the abdomen at laparoscopic examination. Furthermore, the deferent duct was disconnected from the epididymal remnants and the deferent duct was visible at the caudal aspect of the bladder, traversing over the lateral ligament of the bladder into the vaginal ring where it ended blindly in the vaginal process. After laparoscopic removal of the testicular/epididymal remnants and castration of the contralateral normally descended testicle, the hCG stimulation test was negative and the pony no longer exhibited masculine behaviour.
The importance of the equine thoracolumbar vertebral column in orthopaedic disorders is well reco... more The importance of the equine thoracolumbar vertebral column in orthopaedic disorders is well recognized and diagnostic imaging becomes more feasible, but little is known about variations in the anatomical configuration within breeds. In this descriptive post-mortem study, anatomical variations in three widely differing breeds: Warmblood horses, Shetland ponies and semi-feral Konik horses are described. The caudal cervical (C), thoracic (T), lumbar (L) and sacral (S) regions of the vertebral column of 30 Warmblood horses, 29 Shetland ponies and 18 Konik horses were examined using computed tomography and visualized by volume rendering. Homologous/ morphologic variations in the caudal cervical area were frequently seen in Warmblood horses (43%), which was significantly more than in the other breeds (p < 0.001). The as standard described equine formula of 18 T, 6 L and 5 S vertebrae was seen in 78% of Konik horses, but only in 53% Warmblood horses and 38% Shetland ponies, which was significantly different (p < 0.05). Overall, Shetland ponies showed a higher tendency of thoracoization, lumbarization and more variations in the number of vertebrae and pairs of ribs. Ankylosed intertransverse joints (ITJs) between transverse processes of the lumbar vertebrae were most common between the second last and last lumbar vertebra and prevalence was significantly higher in Shetland ponies (61%), than in Warmblood horses (38%) and Konik horses (7%) (p < 0.0001). Cranial to the second last lumbar vertebra there were fewer ITJs ankylosed (14%) in Warmblood horses (p < 0.0095), and this decrease in number of ankylosed ITJs was different compared to the change in ankylosed ITJs in Shetland ponies (p < 0.005). ITJs occurred asymmetrically in 15% (12/77) of the cases. A limitation of the study was that clinical data of the horses were only incompletely available, precluding any conclusions about the potential clinical implications of anatomical variations. Knowledge of variation in osseous anatomy of the equine thoracolumbar vertebral column is important for the interpretation of diagnostic imaging. To assess the functional importance and clinical relevance of this variation, follow-up studies are necessary.
Objective-To evaluate an arthroscopic indentation instrument (Artscan 200) for assessment of the ... more Objective-To evaluate an arthroscopic indentation instrument (Artscan 200) for assessment of the health status of equine articular cartilage. Study Design-In vitro experiment using equine isolated proximal phalanx (P1) specimens. Sample Population-P1 specimens from 39 horses (aged 1.5-22 years). Methods-Reproducibility was tested by determination of the coefficient of variation (CV). Dynamic modulus and cartilage degeneration index (CDI) values were measured at 2 predefined sites (site 1, joint margin; site 2, joint center) to assess the accuracy and to evaluate the relation with surface integrity. Results-CV was 9.0%. A significant decrease in indenter force was identified when dynamic modulus values decreased to o2.5 MPa (range of tested samples 0.9-8.1 MPa) and when CDI values at site 1 increased to 450% (range 5.4-72.8%). Conclusions-Technique reproducibility was adequate but accuracy was limited. The device identified degeneration-associated decreases in cartilage stiffness only when the mechanical properties of the cartilage were considerably changed. Clinical Relevance-Usefulness of this indentation instrument during arthroscopic surgery would be limited in the initial phase of OA-like cartilage degeneration, but may yield important information in more advanced OA.
for performing study: Osteoarthritis (OA) is one of the most prevalent and disabling chronic cond... more for performing study: Osteoarthritis (OA) is one of the most prevalent and disabling chronic conditions affecting horses and leads to degeneration of articular cartilage. Diagnosis is based on clinical signs in combination with radiography, which is relatively insensitive and provides only an indication of accumulated damage. Alternative methods, such as molecular markers, are therefore needed that can quantitatively, reliably and sensitively detect osteoarthritic changes in the joints at an early stage of the disease. If such markers are to be used reliably, it is important to know the relationship between marker concentration and cartilage composition. Objectives: To study the relationship between cartilage composition, synovial fluid levels of glycosaminoglycans (GAGs), hydroxyproline (Hyp) and general matrix metalloproteinase (MMP) activity, and the presence and severity of articular cartilage damage on the articular surface of P1. Methods: Synovial fluid (SF) was collected from the metacarpophalangeal joints of 60 mature horses, and levels of GAGs, Hyp and general MMP activity were determined. Further, GAG and denatured collagen content of the articular cartilage were determined at the dorsal articular margin of P1 (site 1) and central cavity (site 2). The presence and severity of cartilage change was quantified using the cartilage degeneration index (CDI), measured at the same 2 sites. Correlations between SF parameters, cartilage composition and degree of cartilage degeneration were sought using correlation analysis. Results: There was no correlation between GAG or Hyp content of SF and the amount of GAGs or denatured collagen, respectively, in cartilage. In joints with moderate to severe cartilage damage, the GAG content of site 1 was significantly lower than in joints with no to minimal cartilage change (P = 0.005) and there was a negative correlation between the amount of denatured collagen and GAG content at site 1 in all joints (r = -0.39, P = 0.002). Further, in joints with moderate to severe cartilage damage, there was a significant positive correlation between MMP activity in SF and Hyp levels in SF (r = 0.72, P<0.001) and CDI at sites 1 (r = 0.46, P=0.03) and 2 (r = 0.43, P = 0.04). Conclusions: General MMP activity in joints with moderate to severe cartilage damage is related to the severity of those cartilage changes and to Hyp levels in SF. Glycosaminoglycan levels in SF are not directly related to MMP activity, GAG content of articular cartilage or severity of cartilage change. Potential relevance: Glycosaminoglycan levels in SF are not helpful for the early detection of cartilage lesions. In damaged joints, Hyp levels may give an indication of the severity of cartilage change as they are strongly related to MMP activity, but do not qualify as markers for the presence or absence of cartilage lesions.
for performing study: Training at a very young age may influence the characteristics of the colla... more for performing study: Training at a very young age may influence the characteristics of the collagen network of articular cartilage extracellular matrix (ECM) in horses. Objectives: To investigate whether increasing workload of foals results in significant changes in the biochemical composition of articular cartilage ECM. Methods: Thoroughbred foals (n = 33) were divided into 2 different exercise groups from age 10 days-18 months. One group (PASTEX; n = 15) was reared at pasture; the other (CONDEX; n = 18) underwent a specific additional training programme that increased workload by 30%. At mean age 18 months, 6 animals from each group were subjected to euthanasia. The proximal articular surface of the proximal phalanx of the right hindlimb was examined for the presence of damage using the cartilage degeneration index (CDI). Samples were taken from 2 sites with known different loading patterns. Slices were analysed for DNA, glycosaminoglycans (GAG), collagen and post translational modifications of collagen (formation of hydroxylysylpyridinoline [HP] and pentosidine crosslinks, and hydroxylysine [Hyl]), and exercise groups and different sites compared. Results: There were no differences in CDI between PASTEX and CONDEX animals, indicating the absence of extra joint damage due to the exercise regimen. There were site-related differences for most biochemical variables, corroborating earlier reports. All biochemical variables showed differences between PASTEX and CONDEX groups at one of the sites, and some at both. GAG and collagen levels were lower in the CONDEX group whereas Hyl, HP crosslinks and pentosidine crosslinks were higher. Conclusions and potential relevance: A measurable effect of the conditioning exercise was demonstrated. The margin between too much and too little work when training foals may be narrower than intuitively presumed.
Records of lame horses (2004-2007) with (peri-)synovial pain of the metacarpophalangeal joint (MC... more Records of lame horses (2004-2007) with (peri-)synovial pain of the metacarpophalangeal joint (MCPJ, n = 53), digital flexor tendon sheath (DFTS, n = 20), and distal interphalangeal joint (DIPJ, n = 31) were evaluated (n = 104). Cases were grouped according to the following treatments: (1) systemic administration of NSAIDs (n = 40); (2) intrasynovial administration of glucocorticosteroids (n = 30); (3) combination treatment (n = 34). Outcome was defined as successful when the owner reported that the horse had returned to its previous level of performance within a period of up to six months after treatment. Statistical evaluation was performed using Fisher&#39;s Exact tests and multivariable logistic regression analyses at P &lt; 0.05. Successful outcome was significantly higher for horses in treatment group 3 (19/34, 55.9 per cent, P = 0.021) than in treatment groups 1 (11/40, 27.5 per cent, OR 0.21, 95 per cent CI 0.06 to 0.71) and 2 (8/30, 26.6 per cent, OR 0.18, 95 per cent CI 0.06 to 0.56). A significant association was only found with regard to the affected synovial structure (P = 0.025); relative to the DIPJ, involvement of the MCPJ and DFTS was four to five times more likely to provide a successful outcome (OR 4.18 and 5.59, 95 per cent CI 1.24 to 14.08 and 1.42-22.22, respectively).
Arthroscopy enables direct inspection of the articular surface, but provides no information on de... more Arthroscopy enables direct inspection of the articular surface, but provides no information on deeper cartilage layers. Optical coherence tomography (OCT), based on measurement of reflection and backscattering of light, is a diagnostic technique used in cardiovascular surgery and ophthalmology. It provides cross-sectional images at resolutions comparable to that of low-power microscopy. The aim of this study was to determine if OCT is feasible for advanced clinical assessment of lesions in equine articular cartilage during diagnostic arthroscopy. Diagnostic arthroscopy of 36 metacarpophalangeal joints was carried out ex vivo. Of these, 18 joints with varying degrees of cartilage damage were selected, wherein OCT arthroscopy was conducted using an OCT catheter (diameter 0.9 mm) inserted through standard instrument portals. Five sites of interest, occasionally supplemented with other locations where defects were encountered, were arthroscopically graded according to the International Cartilage Repair Society (ICRS) classification system. The same sites were evaluated qualitatively (ICRS classification and morphological description of the lesions) and quantitatively (measurement of cartilage thickness) on OCT images. OCT provided high resolution images of cartilage enabling determination of cartilage thickness. Comparing ICRS grades determined by both arthroscopy and OCT revealed poor agreement. Furthermore, OCT visualised a spectrum of lesions, including cavitation, fibrillation, superficial and deep clefts, erosion, ulceration and fragmentation. In addition, with OCT the arthroscopically inaccessible area between the dorsal MC3 and P1 was reachable in some cases. Arthroscopically-guided OCT provided more detailed and quantitative information on the morphology of articular cartilage lesions than conventional arthroscopy. OCT could therefore improve the diagnostic value of arthroscopy in equine orthopaedic surgery.
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Papers by Harold Brommer