Maturation of the equine medial femoral condyle osteochondral unit.

Objective: The juvenile equine medial femoral condyle (MFC) is frequently affected with radiographic changes (sclerosis and subchondral lucencies) that arise at a similar site to juvenile osteochondritis dissecans (JOCD) in children. There is little information on maturation of the MFC. To describe the normal development of the equine MFC osteochondral unit from birth to 2 years. Methods: Micro CT, histology and immunohistochemistry were performed on healthy equine MFCs (n = 29) at sites where lesions occur. Parameters assessed included: cartilage thickness; the epiphyseal growth plate cartilage organization; the osteochondral junction and progression of endochondral ossification. Results: From 0 to 6 months, chondrocytes near the articular surface are small and flat and have a characteristic hypertrophic appearance near the osteochondral junction but are not arranged in columns like physeal growth plates. The osteochondral junction is also crossed by cartilage canals containing vessels giving a porous appearance on 3D µCT images. At 7 months of age, a subchondral bone plate compact structure emerged histologically coincident with the end of endochondral ossification (absence of type X collagen immunostain and chondrocyte hypertrophy). Conclusion: New information is provided on MFC osteochondral unit maturation that will improve our understanding of the development of juvenile equine orthopaedic disease. Equine MFC endochondral ossification is complete at 6 months of age. The immature osteochondral junction may be structurally fragile because of its microarchitecture and susceptible to focal traumatic events that induce developmental lesions.

Short link N acts as a disease modifying osteoarthritis drug.

Osteoarthritis (OA) is a degenerative joint disease characterised by a progressive degradation of articular cartilage and underlaying bone and is associated with pain and disability. Currently, there is no medical treatment to reverse or even retard OA. Based on our previous reports, where we establish the repair potential of short Link N (sLN) in the intervertebral disc, a cartilage-like tissue, we hypothesise that sLN may hold similar promises in the repair of articular cartilage. This study aimed to determine if sLN, could prevent OA disease progression. Skeletally mature New Zealand white rabbits underwent unilateral anterior cruciate ligament transection (ACLT) of their left femorotibial joints to induce joint degeneration typical of OA. Beginning 3 weeks post-operatively, and every three weeks thereafter for 12 weeks, either saline (1 mL) or sLN (100 µg in 1 mL saline) was injected intraarticularly into the operated knee. Six additional rabbits underwent sham surgery but without ACLT or post-operative injections. The effects on gross joint morphology and cartilage histologic changes were evaluated. In the Saline group, prominent erosion of articular cartilage occurred in both femoral condyle compartments and the lateral compartment of the tibial plateau while, sLN treatment reduced the severity of the cartilage damage in these compartments of the knee showing erosion. Furthermore, statistically significant differences were detected between the joint OA score of the saline and sLN treated groups (p = 0.0118). Therefore, periodic intraarticular injection of sLN is a promising nonsurgical treatment for preventing or retarding OA progression, by reducing cartilage degradation.

Differences in third metacarpal trabecular microarchitecture between the parasagittal groove and condyle at birth and in adult racehorses.

BACKGROUND: The aetiology of equine metacarpal condylar fractures is not completely understood and a developmental cause has been postulated. OBJECTIVES: To investigate the subchondral bone trabecular microarchitecture of the lateral parasagittal groove and condyle in equine neonates and its adaptation with maturation and athletic activity. STUDY DESIGN: Ex vivo observational study. METHODS: Distal metacarpi of neonates, yearlings and adult racehorses (n = 24) were harvested. Dorsal and palmar frontal histological sections, containing the lateral parasagittal groove and condyle, were studied. The sections were digitalised and subchondral trabecular bone quantity and quality parameters and trabecular orientation in the frontal plane were measured. RESULTS: Trabecular spacing and length were greater (P = 0.004 and P = 0.0005 respectively) whereas bone fraction, trabecular number and connectivity were all lower (P = 0.0004, P = 0.0001 and P = 0.001 respectively) in the lateral parasagittal groove compared with the condyle in neonatal foals. Trabecular thickness and bone fraction increased with age in racehorses and trabecular spacing decreased. The predominant trabecular orientation had a consistent pattern in neonates and it changed with maturity and the cumulative effect of racing at all the ROIs except for the palmar lateral parasagittal groove that retained a more 'immature' pattern. MAIN LIMITATIONS: Samples were investigated in 2D. 3D processing could have provided more information. CONCLUSIONS: Already at birth there are striking differences in the subchondral bone trabecular microarchitecture between the lateral parasagittal groove and condyle in foals. Adaptation of trabeculae is confirmed with maturity in racehorses, with the greatest adaptation measured in bone quantity parameters. The trabecular orientation had a unique and more immature orientation pattern in the lateral palmar parasagittal grooves in adult racehorses and may reflect a weaker structure at this site.

Synovial nerve fiber density decreases with naturally-occurring osteoarthritis in horses.

OBJECTIVE: To measure the nerve fiber density in synovial membranes from healthy and OA equine joints and to investigate the relationship between synovial innervation and OA severity, synovial vascularity and synovitis. DESIGN: Twenty-five equine metacarpophalangeal joints were collected post-mortem. The joints were dissected and the macroscopic lesions of the articular cartilage were scored. Synovial membrane specimens (n = 50) were harvested, fixed, sectioned and scored histologically. Immunohistochemical staining and immunofluorescence with S-100 protein, that identifies nerve fibers, and ⍺-actin, that stains vascular smooth muscle, were also performed on site-matched specimens and the relationships between these tissues was interrogated. RESULTS: The nerve fiber density was higher in the superficial layer (≤200 µm) of the synovium when compared to the deeper layer in control equine joints (mean difference (95% C.I.): 0.054% (0.018%, 0.11%)). In osteoarthritic joints, synovial innervation decreased in the superficial layer with increasing macroscopic OA score (ß (SEM), 95% C.I.: -0.0061 (0.00021), -0.0011, -0.00017). The blood vessel density was also higher in the superficial layer of the synovium compared to the deep layer in the control (mean difference (95% C.I.): 1.1% (0.36%, 2.3%)) and OA (mean difference (95% C.I.): 0.60% (0.22%, 1.2%)) equine joints. Moreover, considering all synovial specimens, higher nerve fiber density in the deep layer positively correlated with blood vessel density (ß (SEM), 95% C.I.: 0.11 (0.036), 0.035, 0.18). CONCLUSION: The reduction in nerve fiber density with advanced cartilage degeneration suggests that peripheral neuropathy is associated with equine OA. Whether this link is associated with neuropathic pain, requires further investigation.

Ultrasonographic screening for subclinical osteochondrosis of the femoral trochlea in foals (28-166 days old): a prospective farm study.

BACKGROUND: Extensive osteochondritis dissecans (OCD) lesions of the lateral ridge of the trochlea of the femur (LRTF), the most common OCD-affected site in the stifle, have a poor outcome with surgical debridement and can be career ending. The early detection of osteochondrosis lesions and their conservative management holds the promise to enhance outcome. We hypothesise that ultrasonography is a valuable field screening tool to detect and monitor early subclinical LRTF osteochondrosis. OBJECTIVES: The goals were to 1) describe the normal ultrasonographic features of the LRTF in foals of different ages and 2) screen a foal cohort at the farm for early subclinical osteochondrosis lesions. STUDY DESIGN: Prospective cohort study. METHODS: The LRTF of both hindlimbs of Thoroughbred foals (n = 46, 27-166 days old) were imaged once with ultrasonography and radiography (lateromedial and caudolateral-craniomedial oblique views). Cartilage thickness, ossification front indentation of the chondro-osseous junction and epiphyseal vascularisation were assessed on ultrasonography. Follow-up radiographs were taken as yearlings. RESULTS: The cartilage thickness, ossification front indentation and epiphyseal vascularisation significantly decreased with advancing maturity. Subclinical osteochondrosis lesions, characterised by semicircular indentations in the ossification front (indirect evidence of focal failure of ossification and retained cartilage) were detected in six foals (28-145 days old), both with radiography and ultrasonography. Ultrasonography provided a better overall subjective assessment of the osteochondrosis lesion topography (length, depth and the width) compared with radiography. MAIN LIMITATIONS: Post-mortem validation of lesions was not possible. CONCLUSION: Ultrasonography of the LRTF is a practical, inexpensive and reliable technique to discriminate physiological from pathological events at the LRTF in young foals. It revealed the complex topography of the chondro-osseous junction permitting a rapid, comprehensive assessment of the subclinical osteochondrosis lesions in very young foals.

Equine meniscal degeneration is associated with medial femorotibial osteoarthritis.

BACKGROUND: There is limited information available concerning normal equine meniscal morphology, its degeneration and role in osteoarthritis (OA). OBJECTIVES: To characterise normal equine meniscal morphology and lesions and to explore the relationship between equine meniscal degeneration and femorotibial OA. STUDY DESIGN: Ex vivo cadaveric study. METHODS: Menisci were harvested from 7 normal joints (n = 14 menisci) and 15 joints with OA (n = 30 menisci). A macroscopic femorotibial OA score (cartilage degeneration and osteophytosis) was employed to measure disease severity in each compartment. The femoral and tibial meniscal surfaces were scored for macroscopic fibrillation and tears (1-4). Histological sections (regions: cranial and caudal horn; body) were also scored for microscopic fibrillation and tears (0-3) and inner border degeneration (0-3). RESULTS: Partial meniscal tears were present on both femoral and tibial surfaces in all 3 regions and most frequently identified on the femoral surface of the cranial horn of the medial meniscus and body of the lateral meniscus. There was a significantly positive correlation between the global medial meniscal macroscopic scores and osteophyte (r = 0.7, P = 0.002) or cartilage degeneration (r = 0.5, P = 0.03) scores within the medial femorotibial joint. The global medial meniscal macroscopic score was greater (P = 0.004) in the advanced OA joints compared with control joints. MAIN LIMITATIONS: The menisci were principally from abattoir specimens without a known clinical history because of the challenge in obtaining a large number of specimens with a clinical diagnosis of femorotibial OA. CONCLUSIONS: This study is the first to describe normal equine meniscal morphology and lesions. Meniscal lesions were identified in all segments and on both articular surfaces. Meniscal degeneration significantly correlated with OA severity in the equine medial femorotibial joint. The relationship between OA and meniscal pathology remains to be elucidated.

C2K77 ELISA detects cleavage of type II collagen by cathepsin K in equine articular cartilage.

OBJECTIVES: Develop a species-specific ELISA for a neo-epitope generated by cathepsin K cleavage of equine type II collagen to: (1) measure cartilage type II collagen degradation by cathepsin K in vitro, (2) identify cytokines that upregulate cathepsin K expression and (3) compare cathepsin K with matrix metalloproteinase (MMP) collagenase activity in stimulated cartilage explants and freshly isolated normal and osteoarthritic (OA) articular cartilages. DESIGN: A new ELISA (C2K77) was developed and tested by measuring the activity of exogenous cathepsin K on equine articular cartilage explants. The ELISA was then employed to measure endogenous cathepsin K activity in cultured cartilage explants with or without stimulation by interleukin-1 beta (IL-1ß), tumour necrosis-alpha (TNF-α), oncostatin M (OSM) and lipopolysaccharide (LPS). Cathepsin K activity in cartilage explants (control and osteoarthritic-OA) and freshly harvested cartilage (control and OA) was compared to that of MMPs employing C2K77 and C1,2C immunoassays. RESULTS: The addition of Cathepsin K to normal cartilage caused a significant increase (P < 0.01) in the C2K77 epitope release. Whereas the content of C1,2C, that reflects MMP collagenase activity, was increased in media by the addition to cartilage explants of TNF-α and OSM (P < 0.0001) or IL-1ß and OSM (P = 0.002), no change was observed in C2K77 which also unchanged in OA cartilages compared to normal. CONCLUSIONS: The ELISA C2K77 measured the activity of cathepsin K in equine cartilage which was unchanged in OA cartilage. Cytokines that upregulate MMP collagenase activity had no effect on endogenous cathepsin K activity, suggesting a different activation mechanism that requires further study.

Validation of the ultrasonographic assessment of the femoral trochlea epiphyseal cartilage in foals at osteochondrosis predilected sites with magnetic resonance imaging and histology.

BACKGROUND: Noninvasive imaging tools are needed to screen foal femoropatellar joints to detect subclinical osteochondrosis lesions due to focal failure of endochondral ossification to enhance early management to optimise intrinsic healing events. Recently investigations employing 3T susceptibility-weighted magnetic resonance imaging (3T SWI MRI) and CT have demonstrated their capacity for early osteochondrosis diagnosis, but these technologies are not practical for field screening. We postulate that ultrasonography is a valuable field tool for the detection of subclinical osteochondrosis lesions. OBJECTIVES: The goals were to 1) describe the ultrasonographic features of the femoral trochlea of healthy and osteochondrosis-predisposed neonatal foals, 2) validate the capacity of ultrasound to assess cartilage canal vascular archictecture and the ossification front and 3) evaluate field feasibility in a pilot study. STUDY DESIGN: Experimental study. METHODS: Ultrasonographic evaluation of osteochondrosis predisposed (n = 10) and control (n = 6) femoral trochleas was performed ex vivo and compared with site-matched histological sections and 3T SWI MRI. The articular and epiphyseal cartilage thickness, ossification front indentation and cartilage canal vascular archictecture were assessed at each ROI. Femoral trochleae of foals (n = 3) aged ≈ 1, 3 and 6 months were also evaluated with ultrasonography in field. RESULTS: Ultrasonographic measurements strongly correlated with the histological measurements. There was no difference in the cartilage thickness or ossification front indentation between control and osteochondrosis-predisposed specimens. The cartilage canal vascular archictecture on ultrasonograms corresponded with the vessel pattern observed on site matched histology and 3T SWI MRI. MAIN LIMITATIONS: The number of specimens for study was limited and no early osteochondrosis lesions were present within the predilected group, but a field study is now underway. CONCLUSION: Ultrasonographic examination of the femoral trochlea permitted accurate evaluation of cartilage thickness, cartilage canal vascular archictecture and ossification front indentation in young foals and is a promising, practical tool for screening subclinical osteochondrosis and monitoring and managing lesions at important clinical sites.

Early cathepsin K degradation of type II collagen in vitro and in vivo in articular cartilage.

OBJECTIVE: To characterize the initial events in the cleavage of type II collagen mediated by cathepsin K and demonstrate the presence of the resulting products in human and equine articular osteoarthritic cartilage. DESIGN: Equine type II collagen was digested with cathepsin K and the cleavage products characterized by mass spectrometry. Anti-neoepitope antibodies were raised against the most N-terminal cleavage products and used to investigate the progress of collagen cleavage, in vitro, and the presence of cathepsin K-derived products in equine and human osteoarthritic cartilage. RESULTS: Six cathepsin K cleavage sites distributed throughout the triple helical region were identified in equine type II collagen. Most of the cleavages occurred following a hydroxyproline residue. The most N-terminal site was within three residues of the previously identified site in bovine type II collagen. Western blotting using anti-neoepitope antibodies showed that the initial cleavages occurred at the N-terminal sites and this was followed by more extensive degradation resulting in products too small to be resolved by SDS gel electrophoresis. Immunohistochemical staining of cartilage sections from equine or human osteoarthritic joints showed staining in lesional areas which was not observed in non-arthritic sites. CONCLUSIONS: Cathepsin K cleaves triple helical collagen by erosion from the N-terminus and with subsequent progressive cleavages. The liberated fragments can be detected in osteoarthritic cartilage and may represent useful biomarkers for disease activity.

Osteoclasts are recruited to the subchondral bone in naturally occurring post-traumatic equine carpal osteoarthritis and may contribute to cartilage degradation.

UNLABELLED: The role of osteoclasts in osteochondral degeneration in osteoarthritis (OA) has rarely been investigated in spontaneous disease or animal models of OA. OBJECTIVE: The objectives of the current study were to investigate osteoclast density and location in post-traumatic OA (PTOA) and control specimens from racehorses. METHOD: Cores were harvested from a site in the equine third carpal bone, that undergoes repetitive, high intensity loading. Histological and immunohistochemical (Cathepsin K and Receptor-activator of Nuclear Factor kappa-ß ligand (RANKL)) stained sections were scored (global and subregional) and the osteoclast density calculated. The cartilage histological scores were compared with osteoclast density and RANKL scores. RESULTS: There was a greater density of osteoclasts in PTOA samples and they were preferentially located in the subchondral bone plate. RANKL scores positively correlated to the scores of cartilage degeneration and the osteoclast density. The relationship between hyaline articular cartilage RANKL score and osteoclast density was stronger than that of the subchondral bone RANKL score suggesting that cartilage RANKL may have a role in recruiting osteoclasts. The RANKL score in the articular calcified cartilage correlated with the number of microcracks also suggesting that osteoclasts recruited by RANKL may contribute to calcified cartilage degeneration in PTOA. CONCLUSION: Our results support the hypothesis that osteoclasts are recruited during the progression of spontaneous equine carpal PTOA by cartilage RANKL, contributing to calcified cartilage microcracks and focal subchondral bone loss.

The effects of three-month oral supplementation with a nutraceutical and exercise on the locomotor pattern of aged horses.

REASONS FOR PERFORMING STUDY: Multiple in vitro studies assessing articular tissues have indicated that glucosamine and chondroitin sulphate may possess anti-inflammatory effects, but little is known of their clinical effects in vivo. Many old horses have stiff joints, which is likely to be attributable to inflammation and therapy with these nutraceutical compounds could improve joint function. OBJECTIVES: To assess the clinical effects of a mixed supplement on the improvement of stiff gait in aged horses. STUDY DESIGN: Randomised, blinded, placebo-controlled study. METHODS: A group of 24 geriatric equids (age 29 ± 4 years; mean ± s.d.) received either 3 months oral supplementation with a test compound (containing glucosamine, chondroitin sulfate and methyl sulfonyl methane), or a placebo. Kinematic outcome criteria (primary: stride length; secondary: carpal flexion, fore fetlock extension and tarsal range of motion) were objectively quantified on a treadmill at a walk and trot before and after treatment. RESULTS: Stride length did not change significantly in the treated horses at the end of the trial. In the control group, carpal flexion and fore fetlock extension were significantly increased (P<0.05). CONCLUSIONS: There were no indications of effect of the supplement on gait characteristics. The observations in the control group may have been due to a habituation or exercise effect. This study does not support the use of a glucosamine/chondroitin sulfate/methyl sulfonyl methane supplement to improve stiff gait in geriatric horses because of the lack of a sizeable effect. The significant changes in gait parameters in the control group may indicate the usefulness of exercise regimens in older horses.

Inflammatory stimuli differentially modulate the transcription of paracrine signaling molecules of equine bone marrow multipotent mesenchymal stromal cells.

OBJECTIVE: Osteoarthritis (OA) is a degenerative disease of joint tissues that causes articular cartilage erosion, osteophytosis and loss of function due to pain. Inflammation and inflammatory cytokines in synovial fluid (SF) contribute to OA progression. Intra-articular (IA) injections of multipotent mesenchymal stromal cells (MSCs) are employed to treat OA in both humans and animals. MSCs secrete paracrine pro-inflammatory and anabolic signaling molecules that promote tissue repair. The objective of this study was to investigate the effects of OASF on the gene expression of paracrine signaling molecules by MSCs. METHODS: The effects of Lipopolysaccharide (LPS) and interleukin (IL)-1ß as well as both normal (N) and osteoarthritis (OA) SF stimulations on the expression of paracrine pro-inflammatory (tumor necrosis factor (TNF)-α, IL-1ß, IL-8), modulatory (IL-6) and anabolic (vascular endothelial growth factor (VEGF), transforming growth factor (TGF)-ß1 and insulin-like growth factor (IGF)-1) signaling molecules by equine bone marrow multipotent mesenchymal stromal cells (eBM-MSCs) was investigated employing reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: In contrast with NSF, OASF significantly up-regulated the expression of VEGF in eBM-MSCs. Both NSF and OASF significantly down-regulated the expression of IL-1ß. LPS and IL-1ß significantly increased the expression of pro-inflammatory cytokines (TNF-α, IL-8 and IL-6; and IL-1ß and IL-8 respectively). DISCUSSION: We conclude that the transcription of paracrine signaling molecules in eBM-MSCs is modulated by SF. Furthermore, OA alters the properties of SF and the response of eBM-MSCs. Finally, the effects of LPS or IL-1ß stimulation are distinct to that observed following stimulations with OASF.

Outcome after arthroscopic treatment of lateral femoral trochlear ridge osteochondrosis in sport horses. A retrospective study of 37 horses.

OBJECTIVES: To determine the short- and long-term outcome for sport horses after arthroscopic treatment of osteochondrosis of the lateral trochlear ridge of the femur. METHODS: A retrospective study was performed using the medical records of horses intended for use as English sport horses. Outcome was obtained through telephone questionnaire. RESULTS: Thirty-seven horses, mainly Warmbloods, underwent arthroscopic surgery for treatment of lateral femoral trochlear ridge osteochondrosis. Short-term outcome revealed that 27 of 37 horses had no complications. Seven horses had postoperative lameness and effusion which eventually resolved in four horses. Long-term outcome was available for 29 horses, of which 19 were performing to full expectations. Five horses were athletic but at a lower level than expected, and five horses were unable to be used. The depth of the lesion was significantly associated with short-term complications of effusion and lameness. The depth and length of the lesion were not associated with the long-term outcome, but involvement of structures other than the lateral trochlear ridge (patella, medial trochlear ridge) was associated with a worse prognosis. CLINICAL SIGNIFICANCE: The prognosis for sport horses following stifle arthroscopy for lateral trochlear ridge is similar to that reported in other studies and lesions at other sites within the stifle joint.

Mathematical Modeling of Viral Zoonoses in Wildlife.

Zoonoses are a worldwide public health concern, accounting for approximately 75% of human infectious diseases. In addition, zoonoses adversely affect agricultural production and wildlife. We review some mathematical models developed for the study of viral zoonoses in wildlife and identify areas where further modeling efforts are needed.

Relationship between cartilage and subchondral bone lesions in repetitive impact trauma-induced equine osteoarthritis.

OBJECTIVE: To correlate degenerative changes in cartilage and subchondral bone in the third carpal bone (C3) of Standardbred racehorses with naturally occurring repetitive trauma-induced osteoarthritis. DESIGN: Fifteen C3, collected from Standardbred horses postmortem, were assessed for cartilage lesions by visual inspection and divided into Control (CO), Early Osteoarthritis (EOA) and Advanced Osteoarthritis (AOA) groups. Two osteochondral cores were harvested from corresponding dorsal sites on each bone and scanned with a micro-computed tomography (CT) instrument. 2D images were assembled into 3D reconstructions that were used to quantify architectural parameters from selected regions of interest, including bone mineral density and bone volume fraction. 2D images, illustrating the most severe lesion per core, were scored for architectural appearance by blinded observers. Thin sections of paraffin-embedded decalcified cores stained with Safranin O-Fast Green, matched to the micro-CT images, were scored using a modified Mankin scoring system. RESULTS: Subchondral bone pits with deep focal areas of porosity were seen more frequently in AOA than EOA but never in CO. Articular cartilage damage was seen in association with a reduction in bone mineral and loss of bone tissue. Histological analyses revealed significant numbers of microcracks in the calcified cartilage of EOA and AOA groups and a progressive increase in the score compared with CO bones. CONCLUSION: The data reveal corresponding, progressive degenerative changes in articular cartilage and subchondral bone, including striking focal resorptive lesions, in the third carpal bone of racehorses subjected to repetitive, high impact trauma.

Gait analysis and pain response of two rodent models of osteoarthritis.

The purpose of this study was to compare the gait parameters recorded on the CatWalk and the mechanical sensitivity with von Frey filaments of two putative models of osteoarthritis over a one month period, and to evaluate the effect of celecoxib on these parameters. Animals underwent either a surgical sectioning of the anterior cruciate ligament with partial medial menisectomy (ACLT+pMMx) to create a joint instability model or received an intra-articular injection of monoiodoacetate (MIA) as a putative inflammatory joint pain model. Animals were assessed for four consecutive weeks and knee joints were then evaluated histologically. Spinal cord lumbar enlargements were harvested for selected neuropeptide analysis (substance P (SP) and calcitonin gene related peptide (CGRP)). With the MIA model, significant changes persisted in selected dynamic gait parameters throughout the study in the injured limb as well as with the von Frey filaments. The ACLT+pMMx model in contrast showed no clear differential response between both hind limb for both gait parameters and pain-related behavior with von Frey filaments occurred only on the last day of the study. Neuropeptide analysis of spinal cord lumbar enlargements revealed a significant increase in CGRP concentration in both models and an increase in SP concentration only in the MIA model. Histological evaluation confirmed the presence of articular cartilage lesions in both models, but they were much more severe in the MIA model. Celecoxib had an effect on all selected gait parameters at the very beginning of the study and had an important alleviating effect on mechanical allodynia. These results suggest that the MIA model may be more appropriate for the evaluation of short term pain studies and that celecoxib may modulate mechanical allodynia through central sensitization mechanisms.

Basic methods in histopathology of joint tissues.

Histological and histochemical methods are important tools in the evaluation of joint tissue samples for degenerative joint diseases, both in humans and in animal models. In this respect, standardized, simple, and reliable techniques are mandatory. This chapter describes five basic staining procedures appropriate for macroscopic (Indian ink) and histologic (HE/hematoxylin - eosin) visualization and scoring of cartilage proteoglycan and collagen content (toluidine blue/safranin O and picrosirius red/Goldner's trichrome).

Histopathology atlas of animal model systems - overview of guiding principles.

Animal model systems represent an important adjunct and surrogate for studies of osteoarthritis (OA) in humans. They provide a means to study OA pathophysiology as well as aid in the development of therapeutic agents and biological markers for diagnosing and prognosing the disease. Thus, it is of great importance for the OA scientific community, both in academic as well as industrial research, to standardize scoring systems for evaluating the OA disease process and to make results between different studies comparable. The task of the histopathology initiative of OARSI was to achieve a consensus of scoring systems for the most important species used in OA animal model research (dog, guinea pig, horse, mouse, rabbit, rat, and sheep/goat), which are presented in the various chapters in this special volume of Osteoarthritis & Cartilage together with extra chapters on basic methodology (histochemistry, statistics, morphometry), the specific terminology and a general discussion of animal models in OA research. Standardized definitions are suggested for basic but essential terms such as "grading" and "staging" in order to promote their consistent use and thereby promote improved understanding and data interpretation across all model systems. Thus, this introductory chapter presents an overview of the guiding principles for assessment of important OA animal model systems. Use of such systems, independently or in conjunction with other systems in parallel, should facilitate comparability of results across animal model studies.

The OARSI histopathology initiative - recommendations for histological assessments of osteoarthritis in the rabbit.

AIM: The primary goal of this body of work is to suggest a standardized system for histopathological assessment of experimental surgical instability models of osteoarthritis (OA) in rabbits, building on past experience, to achieve comparability of studies from different centres. An additional objective is to review methodologies that have been employed in the past for assessing OA in rabbits with particular reference to the surgical anterior cruciate ligament transection (ACLT) model. METHODS: A panel of scientists and clinician-scientists with recognized expertise in assessing rabbit models of OA reviewed the literature to provide a critical appraisal of the methods that have been employed to assess both macroscopic and microscopic changes occurring in rabbit joint tissues in experimental OA. In addition, a validation of the proposed histologic histochemical grading system was performed. RESULTS: The ACLT variant of the surgical instability model in skeletally mature rabbits is the variation most capable of reproducing the entire range of cartilage, synovial and bone lesions recognized to be associated with OA. These lesions can be semiquantitatively graded using macroscopic and microscopic techniques. Further, as well as cartilage lesions, this ACLT model can produce synovial and bone lesions similar to that of human OA. CONCLUSIONS: The ACLT variant of the surgical instability model in rabbits is a reproducible and effective model of OA. The cartilage lesions in this model and their response to therapy can be graded according to an adapted histological and histochemical grading system, though also this system is to some extent subjective and, thus, neither objective nor entirely reproducible.