RESUMEN
OBJECTIVE: Gangliosides, ubiquitously existing membrane components that modulate transmembrane signaling and mediate cell-to-cell and cell-to-matrix interactions, are key molecules of inflammatory and neurological disorders. However, the functions of gangliosides in the cartilage degradation process remain unclear. We investigated the functional role of gangliosides in cartilage metabolism related to osteoarthritis (OA) pathogenesis. DESIGN: We generated knockout (KO) mice by targeting the ß1, 4-N-acetylgalactosaminyltransferase (GalNAcT) gene, which encodes an enzyme of major gangliosides synthesis, and the GD3 synthase (GD3S) gene, which encodes an enzyme of partial gangliosides synthesis. In vivo OA and in vitro cartilage degradation models were used to evaluate the effect of gangliosides on the cartilage degradation process. RESULTS: The GalNAcT and GD3S KO mice developed and grew normally; nevertheless, OA changes in these mice were enhanced with aging. The GalNAcT KO mice showed significantly enhanced OA progression compared to GD3S mice in vivo. Both GalNAcT and GD3S KO mice showed severe IL-1α-induced cartilage degradation ex vivo. Phosphorylation of MAPKs was enhanced in both GalNAcT and GD3S KOs after IL-1α stimulation. Gangliosides modulated by GalNAcT or GD3S rescued an increase of MMP-13 induced by IL-1α in mice lacking GalNAcT or GD3S after exogenous replenishment in vitro. CONCLUSION: These data show that the deletion of gangliosides in mice enhanced OA development. Moreover, the gangliosides modulated by GalNAcT are important for cartilage metabolism, suggesting that GalNAcT is a potential target molecule for the development of novel OA treatments.
Asunto(s)
Artritis Experimental/metabolismo , Cartílago Articular/metabolismo , Gangliósidos/fisiología , Osteoartritis/metabolismo , Envejecimiento/fisiología , Animales , Artritis Experimental/patología , Cartílago Articular/efectos de los fármacos , Cartílago Articular/patología , Células Cultivadas , Condrocitos/efectos de los fármacos , Condrocitos/metabolismo , Progresión de la Enfermedad , Gangliósidos/deficiencia , Gangliósidos/farmacología , Eliminación de Gen , Crecimiento/genética , Interleucina-1alfa/antagonistas & inhibidores , Interleucina-1alfa/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/fisiología , Masculino , Metaloproteinasa 13 de la Matriz/biosíntesis , Ratones Noqueados , N-Acetilgalactosaminiltransferasas/deficiencia , N-Acetilgalactosaminiltransferasas/genética , N-Acetilgalactosaminiltransferasas/fisiología , Óxido Nítrico/metabolismo , Osteoartritis/patología , Sialiltransferasas/deficiencia , Sialiltransferasas/genética , Sialiltransferasas/fisiología , Técnicas de Cultivo de Tejidos , Regulación hacia Arriba/fisiología , Polipéptido N-AcetilgalactosaminiltransferasaRESUMEN
OBJECTIVES: We hypothesized that high-molecular-weight (MW) cross-linked (CL) hyaluronic acid (HA) improves joint lubrication and has an enhanced chondroprotective effect. We examined the histopathological changes and friction coefficients in osteoarthritic knee joints after injecting high-MW CL HA. DESIGN: A bilateral anterior cruciate ligament transection (ACLT) model in 20 Japanese white rabbits was used. From week 5 after transection, low-MW HA (0.8 × 10(6) Da; HA80) or high-MW CL HA (6 × 10(6) Da; HA600) was injected weekly into 10 right knee for 3 weeks; normal saline (NS) was injected into the 10 left knee. A sham operation was undertaken to exclude spontaneous osteoarthritis (OA) in five knees. Results were evaluated with macroscopy, histopathology (Kikuchi's score), biomechanical testing, and rheological assessment of the joint fluid viscoelasticity. Statistical analysis was performed using one-way analysis of variance with a 95% confidence interval (CI) (P < 0.05). RESULTS: The macroscopic findings showed severely damaged cartilage in 30% of the NS group and 20% of the HA80 and HA600 groups and intact cartilage in 100% of the sham group. The histological scores and friction coefficients of the HA600 group were significantly lower than those of the NS group (P = 0.007 and P = 0.002, respectively). Viscoelasticity measurements of the joint fluid showed no significant differences between the three treatment groups. CONCLUSION: High-MW CL HA exerts potential chondroprotective effects and produces superior friction coefficients. Our results suggest that HA600 delays the progression of OA effectively and improves joint lubrication significantly.
Asunto(s)
Artritis Experimental/prevención & control , Cartílago Articular/patología , Ácido Hialurónico/uso terapéutico , Osteoartritis de la Rodilla/prevención & control , Viscosuplementos/uso terapéutico , Animales , Lesiones del Ligamento Cruzado Anterior , Artritis Experimental/patología , Artritis Experimental/fisiopatología , Progresión de la Enfermedad , Evaluación Preclínica de Medicamentos/métodos , Elasticidad , Femenino , Fémur/patología , Fricción , Ácido Hialurónico/administración & dosificación , Ácido Hialurónico/química , Inyecciones Intraarticulares , Lubrificación/métodos , Peso Molecular , Osteoartritis de la Rodilla/patología , Osteoartritis de la Rodilla/fisiopatología , Conejos , Líquido Sinovial/fisiología , Viscosidad , Viscosuplementación/métodos , Viscosuplementos/administración & dosificación , Viscosuplementos/químicaRESUMEN
OBJECTIVE: Glycosphingolipids (GSLs) are ubiquitous membrane components that play a functional role in maintaining chondrocyte homeostasis. We investigated the potential role of gangliosides, one of the major components of GSLs, in osteoarthritis (OA) pathogenesis. DESIGN: Both age-associated and instability-induced OA models were generated using GM3 synthase knockout (GM3S(-/-)) mice. A cartilage degradation model and transiently GM3S-transfected chondrocytes were analyzed to evaluate the function of gangliosides in OA development. The amount of each series of GSLs in chondrocytes after IL-1α stimulation was profiled using mass spectrometry (MS). RESULTS: OA changes in GM3S(-/-) mice were dramatically enhanced with aging compared to those in wild-type (WT) mice. GM3S(-/-) mice showed more severe instability-induced pathologic OA in vivo. Ganglioside deficiency also led to the induction of matrix metalloproteinase (MMP)-13 and ADAMTS-5 secretion and chondrocyte apoptosis in vitro. In contrast, transient GM3S transfection of chondrocytes suppressed MMP-13 and ADAMTS-5 expression after interleukin (IL)-1α stimulation. GSL profiling revealed the presence of abundant gangliosides in chondrocytes after IL-1α stimulation. CONCLUSION: Gangliosides play a critical role in OA pathogenesis by regulating the expression of MMP-13 and ADAMTS-5 and chondrocyte apoptosis. Based on the obtained results, we propose that gangliosides are potential target molecules for the development of novel OA treatments.
Asunto(s)
Artritis Experimental/metabolismo , Cartílago Articular/patología , Gangliósidos/deficiencia , Osteoartritis/metabolismo , Proteínas ADAM/genética , Proteínas ADAM/metabolismo , Proteína ADAMTS5 , Envejecimiento/patología , Animales , Apoptosis/fisiología , Artritis Experimental/patología , Cartílago Articular/efectos de los fármacos , Células Cultivadas , Condrocitos/efectos de los fármacos , Condrocitos/metabolismo , Condrocitos/patología , Progresión de la Enfermedad , Gangliósidos/fisiología , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Interleucina-1alfa/farmacología , Masculino , Metaloproteinasa 3 de la Matriz/genética , Metaloproteinasa 3 de la Matriz/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Osteoartritis/patología , Técnicas de Cultivo de TejidosRESUMEN
Knee arthroscopy is a minimally invasive surgery used in the treatment of intra-articular knee pathology which may cause unintended damage to femoral cartilage. An ultrasound (US)-guided autonomous robotic platform for knee arthroscopy can be envisioned to minimise these risks and possibly to improve surgical outcomes. The first necessary tool for reliable guidance during robotic surgeries was an automatic segmentation algorithm to outline the regions at risk. In this work, we studied the feasibility of using a state-of-the-art deep neural network (UNet) to automatically segment femoral cartilage imaged with dynamic volumetric US (at the refresh rate of 1 Hz), under simulated surgical conditions. Six volunteers were scanned which resulted in the extraction of 18278 2-D US images from 35 dynamic 3-D US scans, and these were manually labelled. The UNet was evaluated using a five-fold cross-validation with an average of 15531 training and 3124 testing labelled images per fold. An intra-observer study was performed to assess intra-observer variability due to inherent US physical properties. To account for this variability, a novel metric concept named Dice coefficient with boundary uncertainty (DSCUB) was proposed and used to test the algorithm. The algorithm performed comparably to an experienced orthopaedic surgeon, with DSCUB of 0.87. The proposed UNet has the potential to localise femoral cartilage in robotic knee arthroscopy with clinical accuracy.
Asunto(s)
Artroscopía/métodos , Cartílago Articular/diagnóstico por imagen , Aprendizaje Profundo , Procedimientos Quirúrgicos Robotizados , Cirugía Asistida por Computador , Adulto , Femenino , Humanos , Masculino , Ultrasonografía/métodos , Adulto JovenRESUMEN
The aim of this study was to examine the effects of chronic running exercise on degenerative-regenerative processes in the hindlimb muscles of dystrophin-deficient mdx mice. The number of large-sized degenerative-regenerative groups (DRGs) was markedly decreased, whereas that of small-sized DRGs was unchanged by exercise. Expression of insulin-like growth factor-1 (Igf1), as well as a myogenic factor MyoD (Myod1), was downregulated in mdx muscles by exercise. The downregulation of Igf1 may well correlate with the decrease in the population of early regenerating fibers, which existed predominantly in DRGs, because IGF-1 was mainly localized in these fibers. Our data indicate that chronic exercise may accelerate the active cycle of degeneration-regeneration in mdx skeletal muscles. This means that mdx skeletal muscles can temporarily cope with work-induced injury by enhancing muscle regeneration and repair, but we speculate that an early decline of IGF-1 will accelerate age-dependent muscle wasting and weakness in the later stage of life in mdx mice.