RESUMEN
BACKGROUND: Strategies for articular cartilage repair need to take into account topographical differences in tissue composition and architecture to achieve durable functional outcome. These have not yet been investigated in the equine stifle. OBJECTIVES: To analyse the biochemical composition and architecture of three differently loaded areas of the equine stifle. We hypothesise that site differences correlate with the biomechanical characteristics of the cartilage. STUDY DESIGN: Ex vivo study. METHODS: Thirty osteochondral plugs per location were harvested from the lateral trochlear ridge (LTR), the distal intertrochlear groove (DITG) and the medial femoral condyle (MFC). These underwent biochemical, biomechanical and structural analysis. A linear mixed model with location as a fixed factor and horse as a random factor was applied, followed by pair-wise comparisons of estimated means with false discovery rate correction, to test for differences between locations. Correlations between biochemical and biomechanical parameters were tested using Spearman's correlation coefficient. RESULTS: Glycosaminoglycan content was different between all sites (estimated mean [95% confidence interval (CI)] for LTR 75.4 [64.5, 88.2], for intercondylar notch (ICN) 37.3 [31.9, 43.6], for MFC 93.7 [80.1109.6] µg/mg dry weight), as were equilibrium modulus (LTR2.20 [1.96, 2.46], ICN0.48 [0.37, 0.6], MFC1.36 [1.17, 1.56] MPa), dynamic modulus (LTR7.33 [6.54, 8.17], ICN4.38 [3.77, 5.03], MFC5.62 [4.93, 6.36] MPa) and viscosity (LTR7.49 [6.76, 8.26], ICN16.99 [15.88, 18.14], MFC8.7 [7.91,9.5]°). The two weightbearing areas (LTR and MCF) and the non-weightbearing area (ICN) differed in collagen content (LTR 139 [127, 152], ICN176[162, 191], MFC 127[115, 139] µg/mg dry weight), parallelism index and angle of collagen fibres. The strongest correlations were between proteoglycan content and equilibrium modulus (r: 0.642; p: 0.001), dynamic modulus (r: 0.554; p < 0.001) and phase shift (r: -0.675; p < 0.001), and between collagen orientation angle and equilibrium modulus (r: -0.612; p < 0.001), dynamic modulus (r: -0.424; p < 0.001) and phase shift (r: 0.609; p < 0.001). MAIN LIMITATIONS: Only a single sample per location was analysed. CONCLUSIONS: There were significant differences in cartilage biochemical composition, biomechanics and architecture between the three differently loaded sites. The biochemical and structural composition correlated with the mechanical characteristics. These differences need to be acknowledged by designing cartilage repair strategies.
INTRODUCTION/CONTEXTE: Les stratégies de réparation du cartilage articulaire doivent tenir compte des différences topographiques en ce qui a trait à la composition et l'architecture des tissues, afin d'obtenir un résultat durable et fonctionnel. Cellesci n'ont pas encore été étudiées chez le grasset équin. OBJECTIFS: Analyser la composition biochimique et l'architecture de trois régions du grasset portant une quantité de poids différente. Nous émettons l'hypothèse que les différences entre régions seront corrélées aux caractéristiques biomécaniques du cartilage. TYPE D'ÉTUDE: Étude ex vivo. MÉTHODES: Trente échantillons ostéochondraux par site ont été récoltés à partir de la lèvre latérale de la trochlée fémorale (LTR), le sillon intertrochléaire distal (DITG) et le condyle fémoral médial (MFC). Ceuxci ont été soumis à des tests biochimiques, biomécaniques et une analyse structurelle. Un modèle linéaire mixte avec localisation comme facteur fixe et cheval comme facteur randomisé a été appliqué. Puis, ont suivi des comparaisons par paires de moyennes estimées avec contrôle du taux de fausses découvertes, pour tester les différences entre les divers sites. Les corrélations entre les paramètres biochimiques et biomécaniques ont été testé par le coefficient de corrélation Spearman. RÉSULTATS: Le contenu en glycosaminoglycans était différent à chacun des sites (moyenne estimée [95% CI] pour LTR 75.4 [64.5, 88.2], pour ICN 37.3 [31.9, 43.6], pour MFC 93.7[80.1109.6]µg/mg matière sèche), tout comme le module d'équilibre (LTR2.20 [1.96, 2.46], ICN0.48 [0.37, 0.6], MFC1.36 [1.17, 1.56] MPa), le module dynamique (LTR7.33 [6.54, 8.17], ICN4.38[3.77, 5.03], MFC5.62[4.93, 6.36] MPa) et la viscosité (LTR7.49[6.76, 8.26], ICN16.99 [15.88, 18.14], MFC8.7 [7.91, 9.5]°). Les deux régions portant du poids (LTR et MFC) et la région ne supportant pas de poids (ICN) diffèrent par rapport à leur contenu en collagène (LTR 139 [127152], ICN176 [162191], MFC 127 [115139] µg/mg matière sèche), à l'index de parallélisle et à l'angle des fibres de collagène. Les corrélations les plus fortes étaient entre le contenu en protéoglycans et le module d'équilibre (r: 0.642; p: 0.001), le module dynamique (r: 0.554; p < 0.001) et le changement de phase (r:−0.675; p < 0.001), et entre l'angle d'orientation du collagène et le module d'équilibre (r:−0.612; p < 0.001), le module dynamique (r:−0.424; p < 0.001) et le changement de phase (r: 0.609;p:<0.001). LIMITES PRINCIPALES: Seulement un échantillon par site a été soumis aux analyses. CONCLUSIONS: Il existe des différences significatives dans la composition biochimique, biomécanique et l'architecture du cartilage entre les trois sites échantillonnés. La composition biochimique et structurelle corrèle avec les caractéristiques mécaniques. Ces différences doivent être prises en compte lors de la création de stratégies de réparation du cartilage.
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Cartílago Articular , Animales , Caballos , Cartílago Articular/química , Rodilla de Cuadrúpedos/química , Proteoglicanos/análisis , Glicosaminoglicanos/análisis , Colágeno/análisis , Fenómenos BiomecánicosRESUMEN
Xian-Xiong-Gu-Kang is composed of Epimedium brevicornu, Ligusticum chuanxiong, Radix clematidis, Cinnamomum cassia, and Fructus xanthii. It is used to treat numbness and pain of limbs. In this study, we developed a method to simultaneously quantify 11 components of Xian-Xiong-Gu-Kang (icarrin, epimedin A, epimedin B, epimedin C, icariside II, chlorogenic acid, ligustilide, senkyunolide A, senkyunolide I, ferulic acid, and cinnamic acid) in rat plasma using ultra-performance liquid chromatography coupled with quadrupole linear ion trap mass spectrometry. Chromatographic separation was performed on an ACQUITY UPLC BEH C18 column using gradient elution with a mobile phase comprising acetonitrile and 0.05% (v/v) formic acid aqueous solution. Mass spectrometry detection was performed using positive and negative electrospray ionization in the multiple reaction monitoring mode. The calibration curves of the 11 constituents were linear, with correlation coefficients > 0.99. The intra- and interday accuracy and precision values were within ±15.0%. The extraction recoveries of the 11 constituents and two internal standards were between 66.05 and 105.40%, and the matrix effects were between 86.74 and 112.86%. Using this method, the pharmacokinetic features of the 11 constituents were elucidated in the plasma of osteoarthritic rats after oral administration of the Xian-Xiong-Gu-Kang extract.
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Cromatografía Liquida/métodos , Medicamentos Herbarios Chinos , Osteoartritis , Espectrometría de Masas en Tándem/métodos , Animales , Cinamatos/sangre , Cinamatos/química , Cinamatos/farmacocinética , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/análisis , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacocinética , Flavonoides/sangre , Flavonoides/química , Flavonoides/farmacocinética , Límite de Detección , Modelos Lineales , Masculino , Osteoartritis/metabolismo , Osteoartritis/patología , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Rodilla de Cuadrúpedos/química , Rodilla de Cuadrúpedos/patologíaRESUMEN
OBJECTIVE: Autologous articular cartilage (AC) harvested for repair procedures of high weight bearing (HWB) regions of the femoral condyles is typically obtained from low weight bearing (LWB) regions, in part due to the lack of non-destructive techniques for cartilage composition assessment. Here, we demonstrate that infrared fiber optic spectroscopy can be used to non-destructively evaluate variations in compositional and mechanical properties of AC across LWB and HWB regions. DESIGN: AC plugs (N = 72) were harvested from the patellofemoral groove of juvenile bovine stifle joints, a LWB region, and femoral condyles, a HWB region. Near-infrared (NIR) and mid-infrared (MIR) fiber optic spectra were collected from plugs, and indentation tests were performed to determine the short-term and equilibrium moduli, followed by gravimetric water and biochemical analysis. RESULTS: LWB tissues had a significantly greater amount of water determined by NIR and gravimetric assay. The moduli generally increased in tissues from the patellofemoral groove to the condyles, with HWB condyle cartilage having significantly higher moduli. A greater amount of proteoglycan content was also found in HWB tissues, but no differences in collagen content. In addition, NIR-determined water correlated with short-term modulus and proteoglycan content (R = -0.40 and -0.31, respectively), and a multivariate model with NIR data was able to predict short-term modulus within 15% error. CONCLUSIONS: The properties of tissues from LWB regions differ from HWB tissues and can be determined non-destructively by infrared fiber optic spectroscopy. Clinicians may be able to use this modality to assess AC prior to harvesting osteochondral grafts for focal defect repair.
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Cartílago Articular/química , Cartílago Articular/fisiología , Soporte de Peso/fisiología , Animales , Fenómenos Biomecánicos/fisiología , Agua Corporal , Bovinos , Proteoglicanos/análisis , Espectroscopía Infrarroja Corta/métodos , Rodilla de Cuadrúpedos/química , Rodilla de Cuadrúpedos/fisiologíaRESUMEN
OBJECTIVE: To investigate the impact of a daily exercise dose on cartilage composition and thickness, by conducting a systematic review of randomized controlled trials (RCTs) involving healthy animals. METHODS: A narrative synthesis of the effect of a daily exercise dose on knee cartilage aggrecan, collagen and thickness was performed. A subset of studies reporting sufficient data was combined in meta-analysis using a random-effects model. Meta-regression analyses were performed to investigate the impact of covariates. RESULTS: Twenty-nine RCTs, involving 64 comparisons, were included. In the low dose exercise group, 21/25 comparisons reported decreased or no effect on cartilage aggrecan, collagen and thickness. In the moderate dose exercise group, all 12 comparisons reported either no or increased effect. In the high dose exercise group, 19/27 comparisons reported decreased effect. A meta-analysis of 14 studies investigating cartilage thickness showed no effect in the low dose exercise group (SMD -0.02; 95% CI -0.42 to 0.38; I2 = 0.0%), large but non-significant cartilage thickening in the moderate dose exercise group (SMD 0.95; 95% CI -0.33 to 2.23; I2 = 72.1%) and non-significant cartilage thinning in the high dose exercise group (SMD -0.19; 95% CI -0.49 to 0.12; I2 = 0.0%). Results were independent of analyzed covariates. The overall quality of the studies was poor because of inadequate reporting of data and high risk of bias. CONCLUSIONS: Our results suggest that the relationship between daily exercise dose and cartilage composition, but not necessarily cartilage thickness, may be non-linear. While we found inconclusive evidence for a low daily dose of exercise, a high daily dose of exercise may have negative effects and a moderate daily dose of exercise may have positive effects on cartilage matrix composition in healthy animals.
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Animales de Laboratorio/fisiología , Cartílago Articular/fisiología , Condicionamiento Físico Animal/fisiología , Rodilla de Cuadrúpedos/fisiología , Agrecanos/análisis , Animales , Perros , Matriz Extracelular/química , Femenino , Cobayas , Masculino , Conejos , Ensayos Clínicos Controlados Aleatorios como Asunto , Ratas , Rodilla de Cuadrúpedos/químicaRESUMEN
Proteoglycan 4 (PRG4) is a mucin-like glycoprotein present in synovial fluid and at the surface of articular cartilage. The objectives of this study were to (1) assess the articular cartilage surface adsorption and in vitro cartilage boundary lubricating ability of full-length recombinant human PRG4 (rhPRG4), and (2) cartilage boundary lubricating ability of purified rhPRG4, both alone and in combination with hyaluronan (HA). rhPRG4 adsorption onto articular cartilage explants was assessed by immunohistochemistry and dot blot. An in vitro cartilage-cartilage friction test was used to assess rhPRG4's cartilage boundary lubricating ability compared to bovine PRG4, and that of purified rhPRG4 both alone and in combination with HA. rhPRG4 was able to adsorb to the articular surface, as well as the cut surface, of cartilage explants. The kinetic coefficient of friction of rhPRG4 was similar to that of PRG4 (p = 0.16) and lower than phosphate-buffered saline (p < 0.05), while that of purified rhPRG4 + HA was significantly lower than rhPRG4 alone (p < 0.05). This study demonstrates that rhPRG4 can adsorb to an intact articular cartilage surface and functions as an effective boundary lubricant, both alone and with HA, and provides the foundation for in vivo evaluation of this clinically relevant full-length rhPRG4 for treatment of osteoarthritis.
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Cartílago Articular/química , Ácido Hialurónico/química , Proteoglicanos/química , Adsorción , Animales , Células CHO , Cartílago Articular/metabolismo , Bovinos , Cricetulus , Humanos , Ácido Hialurónico/metabolismo , Proteoglicanos/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Rodilla de Cuadrúpedos/química , Rodilla de Cuadrúpedos/metabolismoRESUMEN
Rheumatoid arthritis (RA) and osteoarthritis (OA) are serologically and clinically distinctive, but at the local level, both diseases have many molecular pathways in common. In vivo molecular imaging can unravel the local pathologic processes involved in both diseases. In this study, we investigated matrix metalloproteinase (MMP) and cathepsin activity during cartilage destruction, in an RA and an OA mouse model, using biophotonic imaging of substrate-based probes. Mice with collagen-induced arthritis (CIA) or destabilization of the medial meniscus (DMM) were imaged using near-infrared fluorescent probes, activated by several cathepsins or MMPs. Fluorescence signal intensity was compared to synovial gene expression, histology, and cartilage staining of a neoepitope of aggrecan cleaved by MMPs with the amino acids DIPEN. Increased cathepsin and MMP activity was seen during CIA, whereas the DMM model only showed increased MMP activity. DIPEN expression was seen only during CIA. A possible explanation can be differences in gene expressions; MMP3 and -13, known to produce DIPEN neoepitopes, were upregulated in the CIA model, whereas MMP12, known to be involved in elastin degradation and chemokine inhibition, was upregulated in the DMM model. Thus, molecular imaging showed no cathepsin activity at the time of cartilage damage in the DMM model, whereas both cathepsins and MMPs are active in the CIA model during disease progression.
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Artritis Reumatoide/metabolismo , Catepsinas/análisis , Metaloproteinasas de la Matriz/análisis , Imagen Molecular/métodos , Osteoartritis/metabolismo , Animales , Artritis Experimental/metabolismo , Catepsinas/metabolismo , Muerte Celular , Condrocitos/citología , Condrocitos/metabolismo , Colágeno Tipo II/efectos adversos , Colágeno Tipo II/inmunología , Masculino , Metaloproteinasas de la Matriz/metabolismo , Ratones , Ratones Endogámicos C57BL , Rodilla de Cuadrúpedos/química , Rodilla de Cuadrúpedos/metabolismo , Rodilla de Cuadrúpedos/patologíaRESUMEN
OBJECTIVES: To measure the activity of matrix metalloproteinases (MMP)-2 and -9 in synovial fluid from the stifle joints of dogs with cranial cruciate ligament (CrCL) rupture and to compare that to values from contralateral stifle joints and dogs with clinically normal stifle joints. Additionally, the C-reactive protein (CRP) levels were also measured. METHODS: Fourteen large breed dogs with unilateral CrCL rupture and 11 large breed normal dogs were included in this prospective clinical study. Synovial fluid was collected from CrCL-ruptured stifle joints, contralateral clinically normal stifle joints of the same dogs, and stifle joints of normal dogs. Serum was also collected. Synovial fluid activities of MMP-2 and MMP-9 and serum CRP level were measured. RESULTS: The MMP-2 activity in synovial fluid was significantly higher in CrCL-ruptured joints compared to contralateral joints and to stifles from normal dogs. There was no significant difference in activity of MMP-2 in contralateral joints of CrCL-ruptured dogs compared to normal dogs. Both serum CRP level and MMP-9 activity did not differ significantly between the studied conditions. CLINICAL SIGNIFICANCE: It was confirmed that MMP-2 activity is significantly related to CrCL rupture, but there was a failure to demonstrate any significant increase in the contralateral joints compared to the stifle joints of normal dogs. The MMP-2 involvement in progressing CrCL disease still has to be defined.
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Lesiones del Ligamento Cruzado Anterior , Enfermedades de los Perros/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Animales , Ligamento Cruzado Anterior/patología , Biomarcadores/sangre , Proteína C-Reactiva/análisis , Enfermedades de los Perros/sangre , Enfermedades de los Perros/diagnóstico , Perros , Regulación Enzimológica de la Expresión Génica , Metaloproteinasa 2 de la Matriz/sangre , Metaloproteinasa 9 de la Matriz/sangre , Metaloproteinasa 9 de la Matriz/metabolismo , Rotura/veterinaria , Rodilla de Cuadrúpedos/química , Rodilla de Cuadrúpedos/patología , Líquido Sinovial/químicaRESUMEN
This study was made to elucidate the changes in the periarticular connective tissue that can underlie the contracture after spasticity development. Sixteen Wistar rats underwent a spinal cord injury and 16 rats were either sham- or nonoperated. The periarticular connective tissue of the knee joint was assessed with histological, histomorphometric, immunohistochemical, and biochemical analyses. Histological results showed a smaller synovial intima, a dense subintimal and posterior joint capsule without fibrosis, and a disarranged posterior capsule in the spinal cord-injured knees with the flexion contracture. The synovial intima length was shortened only at the posterior capsule. Neither the distribution nor expression of type I and III collagen was affected. Contractures after spinal cord injuries are characterized by synovial intima adhesions. A dense and disarranged capsule may lead to joint stiffness. The alteration of periarticular connective tissues exhibits properties characteristic of the contracture after spasticity development.
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Tejido Conectivo/patología , Contractura/patología , Articulación de la Rodilla/patología , Traumatismos de la Médula Espinal/patología , Rodilla de Cuadrúpedos/patología , Membrana Sinovial/patología , Animales , Colágeno Tipo I/análisis , Colágeno Tipo III/análisis , Tejido Conectivo/química , Contractura/etiología , Modelos Animales de Enfermedad , Femenino , Ratas , Ratas Wistar , Traumatismos de la Médula Espinal/complicaciones , Rodilla de Cuadrúpedos/química , Membrana Sinovial/química , Factores de TiempoRESUMEN
Perlecan is a modular heparan sulphate and/or chondroitin sulphate substituted proteoglycan of basement membrane, vascular tissues and cartilage. Perlecan acts as a low affinity co-receptor for fibroblast growth factors 1, 2, 7, 9, binds connective tissue growth factor and co-ordinates chondrogenesis, endochondral ossification and vascular remodelling during skeletal development; however, relatively little is known of its distribution in these tissues during ageing and development. The aim of the present study was to immunolocalise perlecan in the articular and epiphyseal growth plate cartilages of stifle joints in 2-day to 8-year-old pedigree merino sheep. Perlecan was prominent pericellularly in the stifle joint cartilages at all age points and also present in the inter-territorial matrix of the newborn to 19-month-old cartilage specimens. Aggrecan was part pericellular, but predominantly an extracellular proteoglycan. Perlecan was a prominent component of the long bone growth plates and displayed a pericellular as well as a strong ECM distribution pattern; this may indicate a so far unrecognised role for perlecan in the mineralisation of hypertrophic cartilage. A significant age dependant decline in cell number and perlecan levels was evident in the hyaline and growth plate cartilages. The prominent pericellular distribution of perlecan observed indicates potential roles in cell-matrix communication in cartilage, consistent with growth factor signalling, cellular proliferation and tissue development.
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Cartílago Articular/química , Placa de Crecimiento/química , Proteoglicanos de Heparán Sulfato/análisis , Rodilla de Cuadrúpedos/química , Agrecanos , Animales , Animales Recién Nacidos , Proteínas de la Matriz Extracelular/análisis , Inmunohistoquímica , Lectinas Tipo C , Proteoglicanos/análisis , Ovinos , Factores de TiempoRESUMEN
Type X collagen is a short-chain collagen that is strongly expressed in hypertrophic chondrocytes. In this study, we used an immunohistochemical technique exploiting a prolonged hyaluronidase unmasking of type X collagen epitopes to show that type X collagen is not restricted to calcified cartilage, but is also present in normal canine noncalcified articular cartilage. A 30 degrees valgus angulation procedure of the right tibia was performed in 15 dogs at the age of 3 months, whereas their nonoperated sister dogs served as controls. Samples were collected 7 and 18 months after the surgery and immunostained for type X collagen. The deposition of type X collagen increased during maturation from age 43 weeks to 91 weeks. In the patella, most of the noncalcified cartilage stained for type X collagen, whereas, in the patellar surface of the femur, it was present mainly in the femoral groove close to cartilage surface. In femoral condyles, the staining localized mostly in the superficial cartilage on the lateral and medial sides, but not in the central weight-bearing area. In tibial condyles, type X collagen was often observed close to the cartilage surface in medial parts of the condyles, although staining could also be seen in the deep zone of the cartilage. Staining for type X collagen appeared strongest at sites where the birefringence of polarized light was lowest, suggesting a colocalization of type X collagen with the collagen fibril arcades in the intermediate zone. No significant difference in type X collagen immunostaining was observed in lesion-free articular cartilage between controls and dogs that underwent a 30 degrees valgus osteotomy. In osteoarthritic lesions, however, there was strong immunostaining for both type X collagen and collagenase-induced collagen cleavage products. The presence of type X collagen in the transitional zone of cartilage in the patella, femoropatellar groove, and in tibial cartilage uncovered by menisci suggests that it may involve a modification of collagen fibril arrangement at the site of collagen fibril arcades, perhaps providing additional support to the collagen network.
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Cartílago Articular/química , Colágeno Tipo X/análisis , Rodilla de Cuadrúpedos/química , Animales , Cartílago Articular/metabolismo , Colágeno Tipo X/biosíntesis , Perros , Femenino , Inmunohistoquímica , Coloración y Etiquetado , Rodilla de Cuadrúpedos/metabolismoRESUMEN
OBJECTIVE: To map aggrecan cleavage by matrix metalloproteinases (MMPs) and aggrecanases in normal murine tibial articular cartilage (CBA strain) and in the development of spontaneous osteoarthritis (OA) in the STR/ort mouse and to assess the influence of sex hormone status on these conditions in gonadectomized STR/ort mice. METHODS: The distributions of neoepitopes of aggrecan generated by MMP (VDIPEN) and aggrecanase (NITEGE) cleavage were investigated by immunohistochemistry. RESULTS: VDIPEN neoepitope was detected mainly in the pericellular matrix of deep-zone chondrocytes in normal tibial cartilage from STR/ort and CBA mice. In early OA, VDIPEN immunostaining also localized to the pericellular matrix of chondrocytes at the site of the lesion. With increasing severity of OA lesions, VDIPEN immunostaining was also detected in the interterritorial matrix, close to the site of the lesion. In contrast, NITEGE mapped most strongly to the pericellular matrix of upper-zone chondrocytes in normal tibial cartilage. As with VDIPEN, NITEGE was strongly expressed in the pericellular matrix at the site of early OA lesions. With advancing OA, NITEGE colocalized with VDIPEN in both the pericellular and interterritorial matrices of chondrocytes adjacent to OA lesions and in those of the deep zones. Hormone status did not appear to influence the development of OA or the distribution of aggrecan neoepitopes in STR/ort mice. CONCLUSION: MMP- and aggrecanase-generated neoepitopes map predominantly to different regions in normal murine tibial cartilage. However, both groups of enzymes generate increased amounts of neoepitopes in pericellular and interterritorial matrix adjacent to histopathologic lesions of OA. Aggrecan degradation and the development of OA appear to be independent of sex hormone status in this model.
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Cartílago Articular/enzimología , Endopeptidasas/metabolismo , Proteínas de la Matriz Extracelular , Metaloproteinasas de la Matriz/metabolismo , Osteoartritis de la Rodilla/enzimología , Proteoglicanos/metabolismo , Agrecanos , Animales , Cartílago Articular/citología , Cartílago Articular/patología , Condrocitos/química , Condrocitos/citología , Condrocitos/patología , Epítopos/análisis , Matriz Extracelular/química , Técnica del Anticuerpo Fluorescente Indirecta , Lectinas Tipo C , Masculino , Ratones , Ratones Endogámicos CBA , Oligopéptidos/análisis , Osteoartritis de la Rodilla/patología , Ovariectomía , Fragmentos de Péptidos/análisis , Rodilla de Cuadrúpedos/química , Rodilla de Cuadrúpedos/enzimología , Rodilla de Cuadrúpedos/patologíaRESUMEN
OBJECTIVE: To determine the regional composition of water and glycosaminoglycan (GAG) disaccharides of the canine meniscus. SAMPLE POPULATION: 52 menisci from the stifle of dogs. PROCEDURE: Regional sections of each meniscus were weighed, dried, and reweighed to determine water content. Dried tissue specimens were subjected to enzymatic digestion. Analysis and quantification of disaccharide degradation products were performed, using high-performance liquid chromatography. RESULTS: Water content was approximately 65% in polar and central regions of the canine meniscus. Water content of the central region of the lateral meniscus was significantly higher than that of the medial meniscus (P = 0.0090). Chondroitinase digestion of canine meniscal tissue yielded detectable delta Di-HA, delta Di-4S, and delta Di-6S GAG disaccharides. Disaccharides specific to dermatan sulfate and chondroitin D or E sulfate were not detected. Concentrations of delta Di-4S and delta Di-6S were significantly greater in the lateral central region, compared with the medial central region (P = 0.0005 and 0.0002, respectively). CONCLUSION: Water content and delta Di-4S and delta Di-6S concentrations were significantly lower in the central region of the medial meniscus, compared with the central region of the lateral meniscus. Reduced tissue hydration of the medial central region may have been a direct result of its overall decrease in total GAG content. CLINICAL RELEVANCE: The ability to evaluate subtle differences in tissue GAG composition by analytical measurement of their constituent disaccharides may aid in the understanding of the complex material properties of the normal and diseased meniscus, which may be applied to the study of meniscal healing and biomechanics.
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Agua Corporal/química , Cartílago Articular/química , Disacáridos/análisis , Glicosaminoglicanos/química , Animales , Condroitinasas y Condroitín Liasas , Perros , Glicosaminoglicanos/análisis , Rodilla de Cuadrúpedos/químicaRESUMEN
Morphology and distribution of water and lipid were examined in menisci of growing swine. The broadness of menisci and the size of protruded tissue in the inner anterior portion of menisci varied among animals. Small tears of tissues were frequently observed. Both the thickness and tissue weight were greater in the lateral than in the medial meniscus. Water content and intensity of glycosaminoglycan staining with safranin-O were found to be greatest in the inner one-third of the meniscus, and greater in the middle than in the outer one-third of the meniscus. Lipid content was greater in the outer one-third than in the inner two-thirds of the meniscus.