RESUMO
As latexin has been linked with chondrocyte hypertrophic differentiation it is possible that this protein may also be involved in the mineralization of cartilage in OA. Therefore, we correlated latexin expression with the mineralization marker, alkaline phosphatase and determined the mineral deposition in the articular cartilage by analyzing the Ca/P ratio and the collagen fibrils pattern, during the progression of post-traumatic OA in a rat model. OA was induced by medial meniscectomy and post-surgery exercise for 5, 10, 20 and 45 days. Protein expression in articular cartilage was evaluated by immunofluorescence, histochemistry and Western blot. Minerals and structure of collagen fibrils in the superficial zone of cartilage were analyzed by energy dispersive X-ray spectroscopy (EDX) and atomic force microscopy (AFM) respectively. Protein expression analysis showed time-dependent up-regulation of latexin during OA progression. In the cartilage, latexin expression correlated with the expression and activity of alkaline phosphatase. EDX of the superficial zone of cartilage showed a Ca/P ratio closer to theoretical values for basic calcium phosphate minerals. The presence of minerals was also analyzed indirectly with AFM, as the collagen fibril pattern was less evident in the mineralized tissue. Latexin is expressed in articular cartilage from the early stages of post-traumatic OA; however, minerals were detected after latexin expression was up-regulated, indicating that its activity precedes and remains during the pathological mineralization of cartilage. Thus, our results contribute to the identification of molecules involved in the mineralization of articular chondrocytes.
Assuntos
Antígenos/metabolismo , Cartilagem Articular/metabolismo , Regulação da Expressão Gênica , Osteoartrite/etiologia , Osteoartrite/metabolismo , Animais , Calcinose/patologia , Cálcio/metabolismo , Diferenciação Celular , Condrócitos/metabolismo , Colágeno/química , Progressão da Doença , Matriz Extracelular/metabolismo , Hidrólise , Masculino , Microscopia de Força Atômica , Ratos , Ratos Wistar , Fatores de Tempo , Ferimentos e Lesões/fisiopatologiaRESUMO
The balance between synthesis and degradation of the cartilage extracellular matrix is severely altered in osteoarthritis, where degradation predominates. One reason for this imbalance is believed to be due to the ligation of the α5ß1 integrin, the classic fibronectin (FN) receptor, with soluble FN fragments instead of insoluble FN fibrils, which induces matrix metalloproteinase (MMP) expression. Our objective was to determine whether the lack of α5ß1-FN binding influences cartilage morphogenesis in vivo and whether non-ligated α5ß1 protects or aggravates the course of osteoarthritis in mice. We engineered mice (Col2a-Cre;Fn1RGE/fl), whose chondrocytes express an α5ß1 binding-deficient FN, by substituting the aspartic acid of the RGD cell-binding motif with a glutamic acid (FN-RGE). At an age of 5 months the knee joints were stressed either by forced exercise (moderate mechanical load) or by partially resecting the meniscus followed by forced exercise (high mechanical load). Sections of femoral articular knees were analysed by Safranin-O staining and by immunofluorescence to determine tissue morphology, extracellular matrix proteins and matrix metalloproteinase expression. The articular cartilage from untrained control and Col2a-Cre;Fn1RGE/fl mice was normal, while the exposure to high mechanical load induced osteoarthritis characterized by proteoglycan and collagen type II loss. In the Col2a-Cre;Fn1RGE/fl articular cartilage osteoarthritis progressed significantly faster than in wild type mice. Mechanistically, we observed increased expression of MMP-13 and MMP-3 metalloproteinases in FN-RGE expressing articular cartilage, which severely affected matrix remodelling. Our results underscore the critical role of FN-α5ß1 adhesion as ECM sensor in circumstances of articular cartilage regeneration.
Assuntos
Cartilagem Articular/patologia , Fibronectinas/metabolismo , Integrina alfa5beta1/metabolismo , Osteoartrite/patologia , Regeneração/fisiologia , Animais , Cartilagem Articular/citologia , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Condrócitos/patologia , Modelos Animais de Doenças , Matriz Extracelular/metabolismo , Matriz Extracelular/patologia , Fibronectinas/genética , Humanos , Articulação do Joelho/metabolismo , Articulação do Joelho/patologia , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , Camundongos , Camundongos Transgênicos , Osteoartrite/etiologia , Condicionamento Físico Animal/efeitos adversos , Transdução de SinaisRESUMO
Chronic infection with HCV is a leading cause of cirrhosis, hepatocellular carcinoma and liver failure. One of the least understood steps in the HCV life cycle is the morphogenesis of new viral particles. HCV infection alters the lipid metabolism and generates a variety of microenvironments in the cell cytoplasm that protect viral proteins and RNA promoting viral replication and assembly. Lipid droplets (LDs) have been proposed to link viral RNA synthesis and virion assembly by physically associating these viral processes. HCV assembly, envelopment, and maturation have been shown to take place at specialized detergent-resistant membranes in the ER, rich in cholesterol and sphingolipids, supporting the synthesis of luminal LDs-containing ApoE. HCV assembly involves a regulated allocation of viral and host factors to viral assembly sites. Then, virus budding takes place through encapsidation of the HCV genome and viral envelopment in the ER. Interaction of ApoE with envelope proteins supports the viral particle acquisition of lipids and maturation. HCV secretion has been suggested to entail the ion channel activity of viral p7, several components of the classical trafficking and autophagy pathways, ESCRT, and exosome-mediated export of viral RNA. Here, we review the most recent advances in virus morphogenesis and the interplay between viral and host factors required for the formation of HCV virions.
Assuntos
Hepacivirus/ultraestrutura , Hepatite C/virologia , Vírion/ultraestrutura , Montagem de Vírus/genética , Genoma Viral , Hepacivirus/genética , Hepatite C/genética , Humanos , Gotículas Lipídicas/metabolismo , RNA Viral/genética , Vírion/genética , Replicação Viral/genéticaRESUMO
Animal models have been used to understand the basic biology of osteoarthritis (OA) and have helped to identify new candidate biomarkers for the early diagnosis and treatment of this condition. Small animals cannot sufficiently mimic human diseases; therefore, large animal models are needed. Pigs have been used as models for human diseases because they are similar to humans in terms of their anatomy, physiology and genome. Hence, we analyzed articular cartilage and synovial membrane pathology in miniature Vietnamese pigs after a unilateral partial menisectomy and 20-day exercise regimen to determine if the pigs developed pathological characteristics similar to human OA. Histological and protein expression analysis of articular cartilage from menisectomized pigs revealed the following pathologic changes resembling OA: fibrillation, fissures, chondrocyte cluster formation, decrease in proteoglycan content and upregulation of the OA-associated proteins MMP-3, MMP-13, procaspase-3 and IL-1ß. Moreover, histological analysis of synovial membrane revealed mild synovitis, characterized by hyperplasia, cell infiltration and neoangiogenesis. Pathological changes were not observed in the contralateral joints or the joints of sham-operated pigs. Further studies are required to validate such an OA model; however, our results can encourage the use of pigs to study early stages of OA physiopathology. Based on their similarities to humans, pigs may be useful for preclinical studies to identify new candidate biomarkers and novel treatments for OA.
Assuntos
Cartilagem Articular/patologia , Modelos Animais de Doenças , Meniscos Tibiais/cirurgia , Osteoartrite/patologia , Animais , Western Blotting , Feminino , Imuno-Histoquímica , Masculino , Condicionamento Físico Animal , Suínos , Porco Miniatura , Membrana Sinovial/patologiaRESUMO
The Integrin ß1 family is the major receptors of the Extracellular matrix (ECM), and the synthesis and degradation balance of ECM is seriously disrupted during Osteoarthritis (OA). In this scenario, integrins modify their pattern expression and regulate chondrocyte differentiation in the articular cartilage. Members of the Transforming growth factor beta (Tgf-ß) Superfamily, such as Growth differentiation factor 5 (Gdf-5) and Bone morphogenetic protein 7 (Bmp-7), play a key role in joint formation and could regulate the integrin expression during chondrocyte differentiation and osteoarthritis progression in an experimental OA rat model. Decrease of α5 integrin expression in articular cartilage was related with chondrocyte dedifferentiation during OA progression, while increase of α1, α2, and α3 integrin expression was related with fibrous areas in articular cartilage during OA. Hypertrophic chondrocytes expressed αV integrin and was increased in the articular cartilage of rats with OA. Integrin expression during chondrocyte differentiation was also analyzed in a micromass culture system of mouse embryo mesenchymal cells, micromass cultures was treated with Gdf-5 or Bmp-7 for 4 and 6 days, respectively. Gdf-5 induced the expression of the α5 sub-unit, while Bmp-7 induced the expression of the αV sub-unit. This suggests a switch in signaling for prehypertrophic chondrocyte differentiation towards hypertrophy, where Gdf-5 could maintain the articular chondrocyte phenotype and Bmp-7 would induce hypertrophy. Decrease of Ihh expression during late stages of OA in rat model suggest that the ossification in OA rat knees and endochondral ossification could be activated by Bmp-7 and αV integrin in absence of Ihh. Thus, chondrocyte phenotype in articular cartilage is similar to prehypetrophic chondrocyte in growth plate, and is preserved due to the presence of Indian hedgehog (Ihh), Gdf-5 and α5 integrin to maintain articular cartilage and prevent hypertrophy.
Assuntos
Proteína Morfogenética Óssea 7/metabolismo , Diferenciação Celular , Condrócitos/patologia , Fator 5 de Diferenciação de Crescimento/metabolismo , Integrina alfa5/metabolismo , Integrina alfaV/metabolismo , Osteoartrite/metabolismo , Animais , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Condrócitos/metabolismo , Matriz Extracelular/metabolismo , Fibrose , Proteínas Hedgehog/metabolismo , Integrina beta1/metabolismo , Masculino , Camundongos , Osteoartrite/patologia , Ratos Wistar , Transdução de SinaisRESUMO
After a joint lesion, high-impact exercise is a risk factor for the development of osteoarthritis (OA). The degradation of articular cartilage in OA has been associated with the activation of inflammatory cytokine signaling pathways. However, differences in cytokine expression in healthy and injured cartilage after exercise have not yet been analyzed. We used immunofluorescence and Western blot to study the expression of IL-1ß and IL-10 in the articular cartilage of normal (N), sham-operated (S), and menisectomized (OA) rats subjected or not to high-impact exercise (E) for 3, 6, and 10 days (N, NE, S, SE, and OA groups). Cartilage integrity and proteoglycan content were only affected in the OA groups. Exercise increased the amount of IL-1ß and IL-10 positive chondrocytes in NE and SE groups compared with non-exercised groups (N and S). The expression of IL-1ß was up-regulated over time in the NE and OA groups, although in the late stages the increase was higher in the OA groups. In contrast, the expression of anti-inflammatory IL-10 was low in the OA group, whereas in the NE groups expression levels were higher at each time point analyzed. These results suggest that anti- and pro-inflammatory molecules in the cartilage might be tightly regulated to maintain the integrity of the tissue and that when this equilibrium is broken (when the meniscus is removed), the pro-inflammatory cytokines take over and OA develops.
Assuntos
Cartilagem Articular/citologia , Cartilagem Articular/metabolismo , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Osteoartrite/metabolismo , Condicionamento Físico Animal/fisiologia , Animais , Células Cultivadas , Condrócitos/citologia , Masculino , Osteoartrite/patologia , RNA Mensageiro/metabolismo , Ratos Wistar , Regulação para CimaRESUMO
BACKGROUND: Osteoarthritis (OA) is a chronic degenerative disease of the articular cartilage, and its diagnosis is based on symptoms and radiological signs that are only present in the late stages of the disease. Due to the limitations in diagnosing OA before the onset of symptoms, such as pain, little is known about the molecular mechanisms involved in the pathogenesis of OA. Experimental OA models are often used to study the kinetics of the progression of this disease. In this report, we conducted a proteomic study of osteoarthritic cartilage during the early stages of OA using an experimental rat model. RESULTS: Ten proteins that are differentially expressed under early OA conditions were identified by 2-DE and MALDI-TOF/MS. These proteins mediated many processes, such as glycolysis and energy production (Nme2 and Pnp), cartilage matrix (Col2a1), transcription and protein synthesis (Eef1a1 and DJ-1), signal transduction (CaM and Pebp1), transport (Alb and Hba1), and latexin (Lxn). In addition, changes in Lxn expression in early OA were observed and validated by western blot and immunofluorescence analysis. CONCLUSIONS: The proteins that we identified indicate that energy metabolism, cartilage matrix remodelling, and protective cellular mechanisms are associated with early OA. In addition, latexin expression during the early stages of OA could be implicated in cartilage repair.
RESUMO
We identify changes in the expression and localization of α5, α4, and α2 integrins during osteoarthritis (OA) pathogenesis in a rat experimental model. The changes were concomitant with variations in the extracellular matrix (ECM) content and the increase of metalloproteinases (MMPs) activity during OA pathogenesis, which were analyzed by immunofluorescence and Western blot assays. Our results showed an increased expression of α5 and α2 integrins at OA late stages, which was co-related with changes in the ECM content, as a consequence of the MMPs activity. In addition, this is the first report that has shown the presence of α4 integrin since OA early stages, which was co-related with the loss of proteoglycans and clusters formation. However, at late OA stages, the increased expression of α4 integrin in the middle and deep zones of the cartilage was also co-related with the abnormal endochondral ossification of the cartilage through its interaction with osteopontin. Finally, we conclude that ECM-chondrocytes interaction through specific cell receptors is essential to maintain the cartilage homeostasis. However, due to integrins cell signaling is ligand-dependent; changes in the ECM contents could induce activation of either anabolic or catabolic processes, which limits the reparative capacity of chondrocytes, favoring OA severity.
Assuntos
Integrina alfa2/metabolismo , Integrina alfa4/metabolismo , Integrina alfa5/metabolismo , Osteoartrite do Joelho/metabolismo , Índice de Gravidade de Doença , Animais , Modelos Animais de Doenças , Matriz Extracelular/metabolismo , Fibronectinas/metabolismo , Receptores de Hialuronatos/metabolismo , Masculino , Metaloproteases/metabolismo , Osteoartrite do Joelho/patologia , Proteoglicanas/metabolismo , Ratos , Ratos WistarRESUMO
Dengue is currently one of the most important arthropod-borne diseases, causing up to 25,000 deaths annually. There is currently no vaccine to prevent dengue virus infection, which needs a tetravalent vaccine approach. In this work, we describe the cloning and expression in Escherichia coli of envelope domain III-capsid chimeric proteins (DIIIC) of the four dengue serotypes as a tetravalent dengue vaccine candidate that is potentially able to generate humoral and cellular immunity. The recombinant proteins were purified to more than 85 % purity and were recognized by anti-dengue mouse and human sera. Mass spectrometry analysis verified the identity of the proteins and the correct formation of the intracatenary disulfide bond in the domain III region. The chimeric DIIIC proteins were also serotype-specific, and in the presence of oligonucleotides, they formed aggregates that were visible by electron microscopy. These results support the future use of DIIIC recombinant chimeric proteins in preclinical studies in mice for assessing their immunogenicity and efficacy.
Assuntos
Proteínas do Capsídeo/metabolismo , Vacinas contra Dengue , Vírus da Dengue/classificação , Vírus da Dengue/imunologia , Regulação Viral da Expressão Gênica/fisiologia , Proteínas do Envelope Viral/metabolismo , Antígenos Virais/imunologia , Proteínas do Capsídeo/genética , Clonagem Molecular , Vírus da Dengue/genética , Vírus da Dengue/metabolismo , Escherichia coli , Estrutura Terciária de Proteína , Proteínas Recombinantes/imunologia , Sorotipagem , Proteínas do Envelope Viral/genéticaRESUMO
BACKGROUND: Osteoarthritis (OA) is characterized by degeneration of articular cartilage. Animal models of OA induced are a widely used tool in the study of the pathogenesis of disease. Several proteomic techniques for selective extraction of proteins have provided protein profiles of chondrocytes and secretory patterns in normal and osteoarthritic cartilage, including the discovery of new and promising biomarkers. In this proteomic analysis to study several proteins from rat normal articular cartilage, two-dimensional electrophoresis and mass spectrometry (MS) were used. Interestingly, latexin (LXN) was found. Using an immunohistochemical technique, it was possible to determine its localization within the chondrocytes from normal and osteoarthritic articular cartilage. RESULTS: In this study, 147 proteins were visualized, and 47 proteins were identified by MS. A significant proportion of proteins are involved in metabolic processes and energy (32%), as well as participating in different biological functions including structural organization (19%), signal transduction and molecular signaling (11%), redox homeostasis (9%), transcription and protein synthesis (6%), and transport (6%). The identified proteins were assigned to one or more subcellular compartments.Among the identified proteins, we found some proteins already recognized in other studies such as OA-associated proteins. Interestingly, we identified LXN, an inhibitor of mammalian carboxypeptidases, which had not been described in articular cartilage. Immunolabeling assays for LXN showed a granular distribution pattern in the cytoplasm of most chondrocytes of the middle, deep and calcified zones of normal articular cartilage as well as in subchondral bone. In osteoarthritic cartilage, LXN was observed in superficial and deep zones. CONCLUSIONS: This study provides the first proteomic analysis of normal articular cartilage of rat. We identified LXN, whose location was demonstrated by immunolabeling in the chondrocytes from the middle, deep and calcified zones of normal articular cartilage, and superficial and deep zones of osteoarthritic cartilage.
RESUMO
Although the molecular mechanisms for initiation of cartilage destruction in osteoarthritis (OA) are unknown, it has been demonstrated that disruption of mitogen-inducible gene 6 (Mig-6) in mice leads to the onset of a degenerative joint disease like OA. On this basis, we correlated gene expression of Mig-6 with Wnt-9a and Wnt-7b genes; we showed downregulation of Mig-6, Wnt-7b, and Wnt-9a during OA, while Wnt-7b was expressed also in osteoblast-like cells. Here we suggest that Aggrecan degradation occurs before the downregulation of Mig-6. It remains to be proven whether there is any relation between Wnt signaling and Aggrecan degradation.
Assuntos
Proteínas de Transporte/biossíntese , Regulação da Expressão Gênica , Osteoartrite/metabolismo , Proteínas Wnt/biossíntese , Agrecanas/metabolismo , Animais , Cartilagem/metabolismo , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Hibridização In Situ , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Modelos Biológicos , Ratos , Ratos Wistar , Transdução de SinaisRESUMO
The morphology of the normal human and rat articular cartilage was assessed using transmission electron microscopy (TEM), atomic force microscopy (AFM), and two-photon excitation (2PE) microscopy. Spurr-embedded sections from fixed human cartilage were simultaneously evaluated using TEM and AFM. The presences of tracks among the chondrocytes from the superficial zone of the cartilage were observed. In order to ratify the presence of interconnecting tracks among superficial zone chondrocytes, whole fixed human and rat cartilage, as well as fresh whole rat cartilage, were examined under the 2PE. In all cases, these tracks were observed. In addition, porous matrix, well-defined lacunae, and cytoplasmic projections anchored to the extracellular matrix (ECM) were also detected. We conclude that normal human and rat flattened superficial chondrocytes might be interconnected by tracks running through the ECM. In addition, cytoplasmic projections were observed anchored to the ECM. All these structures may possibly be related to cell/cell and ECM/chondrocytes signaling. Our findings provide new information that possibly will be of relevant importance for a more profound study of normal cartilage physiology and eventually, the pathogenesis of osteoarthritis.
Assuntos
Cartilagem Articular/ultraestrutura , Condrócitos/citologia , Animais , Proteínas da Matriz Extracelular/metabolismo , Humanos , Microscopia de Força Atômica , Microscopia Eletrônica de Transmissão , RatosRESUMO
Entamoeba histolytica is a protozoan parasite causing dysentery and in some cases liver abscesses. These effects have been attributed to cytolytic substances released by exocytosis. In this study, the presence of the proteins syntaxin-1 and SNAP-25, which are assumed to be involved in exocytosis, were examined by immunohistochemistry, immunoelectron microscopy and western blot analysis. Syntaxin-1 and SNAP-25 were expressed in the vesicular, vacuolar and plasma membranes of E. histolytica trophozoites. It can be concluded that these proteins might be involved in exocytosis processes.
RESUMO
Previously we improved the neurotensin (NT)-polyplex by the coupling of HA2 fusogenic peptide (FP) and Vp1 SV40 karyophilic peptide (KP). We now report the proportion of [(125)I]-NT, [(3)H]-FP, and poly-l-lysine (PLL) in the NT-polyplex, and some of its biophysical properties. We concluded that the most efficient NT-polyplex comprised 1 NT, 4 FP, and 2 PLL molecules. Electrophoresis revealed that high acidity is detrimental for NT-polyplex stability. Electron microscopy and electrophoresis studies showed that 6 muM KP and 1% serum condensed the plasmid DNA (pDNA) before the appearance of toroid structures. Four plasmids were used to evaluate the transfection efficiency. In vitro, maximum expression was produced at molar ratios (pDNA : [(125)I]-NT-[(3)H]-FP-PLL conjugate) of 1:34 for pEGFP-N1 and 1:27 for pECFP-Nuc. Cotransfection of those plasmids was attained at their optimum molar ratios. In vivo, maximum expression of the pDAT-BDNF-flag in dopamine neurons was produced at a 1:45 molar ratio, whereas that of pDAT-EGFP was at 1:20. The NT-polyplex in the presence of 1 muM SR-48692, an NT-receptor specific antagonist, and untargeted polyplex did not cause transfection in vivo demonstrating the specificity of gene transfer via NT-receptor endocytosis. This information is essential for synthesizing an efficient NT-polyplex that can provide a useful tool for specific gene transfection.
Assuntos
Técnicas Genéticas , Neurotensina/química , Animais , Biofísica/métodos , Linhagem Celular Tumoral , Dopamina/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Masculino , Neurônios/metabolismo , Sinais de Localização Nuclear , Pirazóis/química , Quinolinas/química , Ratos , Ratos Wistar , TransfecçãoRESUMO
Understanding the mechanism of hepatitis C virus (HCV) pathogenesis is an important part of HCV research. Recent experimental evidence suggests that the HCV core protein (HCcAg) has numerous functional activities. These properties suggest that HCcAg, in concert with cellular factors, may contribute to pathogenesis during persistent HCV infection. HCV is capable of infecting cells other than hepatocytes. Although the extrahepatic cellular tropism of HCV may play a role in the pathophysiology of this infection, the precise biological significance of the presence of HCV components in different liver cell types presently remains to be established. In this study, HCcAg was detected in nonparenchymal liver cells of six patients out of eight positive for serum HCV RNA. Immunostaining with anti-HCcAg mAbs revealed the presence of this protein in different liver cell types such as lymphocytes, Kupffer, polymorphonuclear, pit, endothelial, stellate, and fibroblast-like cells. Interestingly, HCcAg was immunolabeled not only in the cytoplasm but also in the nucleus of these cells. Remarkably, HCcAg co-localized with large lipid droplets present in stellate cells and with collagen fibers in the extracellular matrix. Moreover, HCcAg was immunolabeled in bile canaliculus suggesting the involvement of the biliary system in the pathobiology of HCV. Data suggest that nonparenchymal liver cells may constitute a reservoir for HCV replication. Besides, HCcAg may contribute to modulate immune function and fibrosis in the liver as well as steatosis.
Assuntos
Núcleo Celular/metabolismo , Hepatite C Crônica/metabolismo , Hepatite C Crônica/patologia , Hepatócitos/metabolismo , Hepatócitos/patologia , Proteínas do Core Viral/metabolismo , Adulto , Biópsia , Núcleo Celular/ultraestrutura , Feminino , Hepacivirus/metabolismo , Hepatócitos/ultraestrutura , Humanos , Masculino , Microscopia Eletrônica de Transmissão , Pessoa de Meia-IdadeRESUMO
The aim of this work was to study the ontogeny of chondrocyte cell division using embryo, adult and osteoarthritic (OA) cartilage. We searched for mitosis phases and performed a comparative evaluation of mitotic index, basic fibroblast growth factor b (FGFb), transforming growth factor beta1 (TGF-beta1) receptors, cyclin dependent kinase (CDK1) and Cyclin-B expression in fetal, neonate, 3, 5, 8 weeks old rats and experimental OA. Our results showed that mitosis phases were observed in all normal cartilage studied, although, we found a decrease in mitotic index in relation to tissue development. No mitosis was detected in OA cartilage. We also found a statistical significant reduction in cell number in OA cartilage, compared with the normal tissue. Furthermore, FGFb and TGF-beta1 receptors diminished in relation to tissue development, and were very scarce in experimental OA. Western blot assays showed CDK-1 expression in all cases, including human-OA cartilage. Similar results were observed for Cyclin-B, except for 8 weeks, when it was not expressed. Our results suggest that cell division seems to be scarce, if not absent within the OA cartilage studied. Nevertheless, the existence of factors essential for cell division leaves open the question concerning chondrocyte proliferation in OA cartilage, which is likely to be present in the early stages of the disease.
Assuntos
Cartilagem/embriologia , Cartilagem/crescimento & desenvolvimento , Condrócitos/citologia , Osteoartrite/metabolismo , Receptores de Ativinas Tipo I/metabolismo , Animais , Proteína Quinase CDC2/metabolismo , Cartilagem/citologia , Proliferação de Células , Condrócitos/metabolismo , Ciclina B/metabolismo , Mitose , Proteínas Serina-Treonina Quinases , Ratos , Ratos Wistar , Receptor do Fator de Crescimento Transformador beta Tipo I , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Receptores de Fatores de Crescimento Transformadores beta/metabolismoRESUMO
Nanotechnology is expected to open new avenues to fight and prevent disease using atomic scale tailoring of materials. Among the most promising nanomaterials with antibacterial properties are metallic nanoparticles, which exhibit increased chemical activity due to their large surface to volume ratios and crystallographic surface structure. The study of bactericidal nanomaterials is particularly timely considering the recent increase of new resistant strains of bacteria to the most potent antibiotics. This has promoted research in the well known activity of silver ions and silver-based compounds, including silver nanoparticles. The present work studies the effect of silver nanoparticles in the range of 1-100 nm on Gram-negative bacteria using high angle annular dark field (HAADF) scanning transmission electron microscopy (STEM). Our results indicate that the bactericidal properties of the nanoparticles are size dependent, since the only nanoparticles that present a direct interaction with the bacteria preferentially have a diameter of approximately 1-10 nm.
RESUMO
Chondrocytes are capable of engulfing latex particles, cell detritus, and necrotic and apoptotic remains in vitro. It is conceivable that chondrocytes might be involved in the clearance by phagocytosis of different materials within the cartilage. In fact, so far there is no evidence for the presence of "professional phagocytes" (macrophages and neutrophils) in this tissue. Chondrocyte suspensions obtained from rat knees and hips were cultured to assess phagocytosis of latex particles (1 microm), articular cartilage detritus, and necrotic and apoptotic chondrocyte remains (induced by VP-16 1 mM). We observed that chondrocytes phagocytosed latex particles as evaluated by confocal microscopy and flow cytometry. In addition, we observed that chondrocytes phagocytosed articular cartilage detritus and necrotic and apoptotic VP-16 induced-chondrocytes, as observed by bright field microscopy and transmission electron microscopy.
Assuntos
Cartilagem Articular/fisiopatologia , Condrócitos/fisiologia , Fagocitose/fisiologia , Animais , Apoptose/fisiologia , Condrócitos/ultraestrutura , Citometria de Fluxo , Masculino , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Microesferas , Ratos , Ratos WistarRESUMO
Se sabe que el lavado articular y el debridamiento son opciones en el tratamiento de la osteoartritis de rodilla. Se realizó un estudio prospectivo, abierto y aleatorizado que incluyó 100 pacientes con OA de rodilla (criterio ACR) en estadios II y III de Kellgren y Lawrence para evaluar la utilidad y la eficacia del lavado articular por punción en comparación con el lavado y debridamiento artroscópico. Se distribuyeron en: grupo A (lavado por punción) y grupo B (lavado y debridamiento artroscópico). Se consideraron variables relacionadas con el dolor y la función articular que fueron analizadas al inicio del tratamiento y 90 d después; se realizó un análisis estadístico con el empleo del test de Chi cuadrado y el de t-Student, significación estadística p<0,05. Se observó que ambos grupos de pacientes experimentaron mejoría en todas las variables analizadas con las 2 variantes de tratamiento, sin diferencias estadísticamente significativas entre uno y otro grupo, los pacientes en un 90 por ciento aproximadamente de los casos experimentaron satisfacción con el proceder, sin ninguna complicación. Se concluyó que el lavado articular por punción y el lavado y debridamiento artroscópico mostraron ser útiles en el alivio de los síntomas en pacientes con OA ligera y moderada a los 3 meses de observación(AU)