RESUMO
Osteoarthritis is recognised to be an interactive pathological process involving the cartilage, subchondral bone and synovium. The signals from the synovium play an important role in cartilage metabolism, but little is known regarding the influence of the signalling from bone. Additionally, the collagenases and stromelysin-1 are involved in cartilage catabolism through mitogen-activated protein kinase (MAPK) signalling, but the role of the gelatinases has not been elucidated. Here, we studied the influence of osteoclastic signals on chondrocytes by characterising the expression of interleukin-1β (IL-1β)-induced gelatinases through MAPK signalling. We found that osteoclast-conditioned media attenuated the gelatinase activity in chondrocytes. However, IL-1β induced increased levels of gelatinase activity in the conditioned media group relative to the mono-cultured chondrocyte group. More specifically, IL-1β restored high levels of gelatinase activity in c-Jun N-terminal kinase inhibitor-pretreated chondrocytes in the conditioned media group and led to lower levels of gelatinase activity in extracellular signal-regulated kinase or p38 inhibitor-pretreated chondrocytes. Gene expression generally correlated with protein expression. Taken together, these results show for the first time that signals from osteoclasts can influence gelatinase activity in chondrocytes. Furthermore, these data show that IL-1β restores gelatinase activity through MAPK inhibitors; this information can help to increase the understanding of the gelatinase modulation in articular cartilage.
Assuntos
Animais , Camundongos , Células 3T3 , Cartilagem Articular , Biologia Celular , Sobrevivência Celular , Fisiologia , Células Cultivadas , Condrócitos , Técnicas de Cocultura , Meios de Cultivo Condicionados , Gelatinases , Interleucina-1beta , Farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno , Sistema de Sinalização das MAP Quinases , Fisiologia , Metaloproteinase 2 da Matriz , Metaloproteinase 9 da Matriz , Proteínas Quinases Ativadas por Mitógeno , Monócitos , Biologia Celular , NF-kappa B , Osteoclastos , Fisiologia , Inibidores de Proteases , Inibidor Tecidual de Metaloproteinase-1 , Inibidor Tecidual de Metaloproteinase-2 , Proteínas Quinases p38 Ativadas por MitógenoRESUMO
<p><b>AIM</b>Given the well-known properties of Schwann cells in promoting nerve regeneration, transplanting Schwann cells into implant sockets might be an effective method to promote sensory responses of osseointegrated implants. The aim of this study was to evaluate the interaction between Schwann cells and osteoblasts.</p><p><b>METHODOLOGY</b>Schwann cells derived from the sciatic nerves of neonatal rat were co-culured with osteoblasts using Transwell inserts. The proliferation of Schwann cells in the co-culture system was evaluated using methylthiazol tetrazolium (MTT) colorimetric method. Moreover, the secretions and mRNA levels of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative real-time PCR, respectively. In order to test the effect of Schwann cells on osteoblasts, alkaline phosphatase (ALP) staining and Alizerin red staining were performed as well.</p><p><b>RESULTS</b>Schwann cells, which were co-cultured with the osteoblasts, showed an intact proliferation during the observation period. Moreover, the gene expression and synthesis of BDNF and NGF were not impaired by the osteoblasts. Meanwhile, co-cultured osteoblasts exhibited a significant increase in the proliferation on day 3 and 6 (P< 0.05). Co-culture of these two types of cells also led to a more intense staining of ALP and an elevated number of calcified nodules.</p><p><b>CONCLUSION</b>These findings demonstrate that, in the in vitro indirect co-culture environment, Schwann cells can maintain their normal ability to synthesize neurotrophins, which then enhance the proliferation and differentiation of osteoblasts.</p>
Assuntos
Animais , Ratos , Fator Neurotrófico Derivado do Encéfalo , Genética , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Expressão Gênica , Fator de Crescimento Neural , Genética , Regeneração Nervosa , Osteoblastos , Biologia Celular , Ratos Sprague-Dawley , Células de Schwann , Biologia Celular , MetabolismoRESUMO
<p><b>AIM</b>To investigate the effect of DAPT (gamma-secretase inhibitor) on the growth of human tongue carcinoma cells and to determine the molecular mechanism to enable the potential application of DAPT to the treatment of tongue carcinoma.</p><p><b>METHODOLOGY</b>Human tongue carcinoma Tca8113 cells were cultured with DAPT. Cell growth was determined using Indigotic Reduction method. The cell cycle and apoptosis were analyzed by flow cytometry. Real-time PCR and Immuno-Fluorescence (IF) were employed to determine the intracellular expression levels.</p><p><b>RESULTS</b>DAPT inhibited the growth of human tongue carcinoma Tca8113 cells by inducing G0-G1 cell cycle arrest and apoptosis. The mRNA levels of Hairy/Enhancer of Split-1 (Hes-1), a target of Notch activation, were reduced by DAPT in a dose-dependent manner. Coincident with this observation, DAPT induced a dose-dependent promotion of constitutive Caspase-3 in Tca8113 cells.</p><p><b>CONCLUSION</b>DAPT may have a therapeutic value for human tongue carcinoma. Moreover, the effects of DAPT in tumor inhibition may arise partly via the modulation of Notch-1 and Caspase-3.</p>