Home About us Contact
|
Home About us Contact | ||
|
|
||
Vitro Cell Culture Systems (vitro + cell_culture_system)
Selected AbstractsHypoxia-like effect of Cobalt Chromium alloy micro particles on fibroblasts in vitroJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 10 2010Bernadette K. Madathil Abstract Periprosthetic osteolysis leading to asceptic loosening remains the primary cause of failure of joint replacement. Although many inflammatory cell types have been implicated, the exact pathomechanisms of asceptic loosening have not been delineated. In the present study we have adopted a proteomic approach to elucidate the initial signals that are expressed to particulate material, using an in vitro cell culture system. Human lung fibroblasts MRC-5 were cultured with Cobalt Chromium (CoCr ASTM F-75, 1,7,µm) particles. Cells were harvested after 72,h incubation and total cellular proteins extracted for downstream analysis via 2D Gel Electrophoresis and tandem mass spectrometry using MALDI-TOF-TOF-MS. Thirteen protein spots showed greater than twofold increase, following 72,h incubation of fibroblast with CoCr particles. Four of these proteins were identified by tandem mass spectrometry. These were Annexin II, Pyruvate kinase, Triose phosphate isomerase, and N-myc downstream regulated gene 1 protein. Cobalt is a hypoxia mimicking agent and N-myc downstream regulated gene 1 protein, Triose phosphate isomerase, Pyruvate kinase, and Annexin II are important hypoxia regulated gene products that are found to be over expressed in cellular oxidative stress response. Our data indicates that exposure of fibroblast to CoCr alloy induces the transition of these cells into a hypoxia like state and oxidative stress even in normoxic culture conditions. The study reflects the possibility of the presence of a hypoxic environment in the periprosthetic tissue surrounding metallic implants. Published by Wiley Periodicals, Inc. J Orthop Res 28:1360,1367, 2010 [source] Nanoporous aluminum oxide affects neutrophil behaviourMICROSCOPY RESEARCH AND TECHNIQUE, Issue 5 2004M. Karlsson Abstract This study evaluates neutrophil responses on aluminum oxide membranes. Using an in vitro cell culture system, we have found that the pore size (20 and 200 nm in diameter) of alumina membranes have a significant effect on leukocyte morphology and activation. Specifically, our results show that 20-nm pore-size membranes were more potent in triggering PMN spreading and extending of pseudopodia than 200-nm pore-size membranes. The morphological changes are also associated with cell activation. In fact, adherent neutrophils on 20-nm pore-size membranes elicit much stronger initial oxygen free radical production. Overall, our results point out that membrane pore size significantly affects the extent of cellular responses of adherent neutrophils. Microsc. Res. Tech. 63:259,265, 2004. © 2004 Wiley-Liss, Inc. [source] Anti-allergic effects of PG102, a water-soluble extract prepared from Actinidia arguta, in a murine ovalbumin-induced asthma modelCLINICAL & EXPERIMENTAL ALLERGY, Issue 2 2009D. Kim Summary Background Asthma is a chronic inflammatory disease of the lung and its incidence has been increasing around the world. We previously reported that oral administration of a water-soluble extract prepared from Actinidia arguta, code-named PG102, could modulate the level of Th1 and Th2 cytokines and suppress the production of immunoglobulin E (IgE) in the ovalbumin (OVA)-immunized murine model as well as in the in vitro cell culture system, and furthermore could significantly improve dermatitis conditions in the NC/Nga murine model. These data suggested that PG102 might have therapeutic effects in a broad range of allergic diseases. Objective To assess the possible anti-allergic effects of PG102 in the OVA-induced murine asthma model. Methods The quality of PG102 was standardized, using its effects on the production of IgE, IL-5, and IL-13, in in vitro cell culture systems. To test effects on asthma, BALB/c mice were orally administrated with PG102, followed by OVA sensitization and challenge to induce asthmatic symptoms. Airway hyperresponsiveness (AHR), bronchoalveolar lavage fluid, serum, and lung tissue were analysed by using various methods. Results PG102 could decrease the level of IgE, IL-5, and IL-13 in in vitro cell culture systems with IC50 being 1.12,1.43 mg/mL. PG102 could ameliorate asthmatic symptoms, including AHR and eosinophilia in the lungs. Such improvement of asthmatic symptoms by PG102 was accompanied by the down-regulation of IL-5 and IgE. In PG102-treated mice, high level expression of heme oxygenase-1, a potent anti-inflammatory enzyme, was observed in alveolar inflammatory cells, while the mRNA levels of foxp3, TGF-,1, and IL-10, important markers for regulatory T cells, were also up-regulated in the lung tissue. Conclusions PG102 may have potential as a safe and effective reagent for the prevention or treatment of asthma. [source] Experimental models for hepatitis C viral infection,HEPATOLOGY, Issue 5 2009Andre Boonstra Hepatitis C virus (HCV) infection is a leading cause of chronic liver disease. The majority of infected individuals develop a persistent infection, which is associated with a high risk of liver cirrhosis and hepatocellular carcinoma. Since its discovery 20 years ago, progress in our understanding of this virus has been suboptimal due to the lack of good model systems. However, in the past decade this has greatly accelerated with the development of various in vitro cell culture systems and in vivo small-animal models. These systems have made a major impact on the field of HCV research, and have provided important breakthroughs in our understanding of HCV infection and replication. Importantly, the in vitro cell culture systems and the small-animal models have allowed preclinical testing of numerous novel antiviral compounds for the treatment of chronic HCV infection. In this article, we give an overview of current models, discuss their limitations, and provide future perspectives for research directed at the prevention and cure of hepatitis C. (HEPATOLOGY 2009.) [source] Presenilin function and ,-secretase activityJOURNAL OF NEUROCHEMISTRY, Issue 4 2005A. L. Brunkan Abstract Alzheimer's disease (AD) is the most common form of dementia and is characterized pathologically by the accumulation of ,-amyloid (A,) plaques and neurofibrillary tangles in the brain. Genetic studies of AD first highlighted the importance of the presenilins (PS). Subsequent functional studies have demonstrated that PS form the catalytic subunit of the ,-secretase complex that produces the A, peptide, confirming the central role of PS in AD biology. Here, we review the studies that have characterized PS function in the ,-secretase complex in Caenorhabditis elegans, mice and in in vitro cell culture systems, including studies of PS structure, PS interactions with substrates and other ,-secretase complex members, and the evidence supporting the hypothesis that PS are aspartyl proteases that are active in intramembranous proteolysis. A thorough knowledge of the mechanism of PS cleavage in the context of the ,-secretase complex will further our understanding of the molecular mechanisms that cause AD, and may allow the development of therapeutics that can alter A, production and modify the risk for AD. [source] Percutaneous permeation of enantiomers and racemates of chiral drugs and prediction of their flux ratios using thermal data: A pharmaceutical perspectiveCHIRALITY, Issue 5 2003Mohsen I. Afouna Abstract Albeit pharmacological, pharmacokinetic, and toxicological differences between enantiomeric pairs or between the pure enantiomers and racemate of chiral drugs are known to exist for decades, we are just beginning to realize that there are apparent differences between these species with respect to their percutaneous permeation as well. Such differences in permeation are likely to be enhanced when chiral drugs are formulated with chiral excipients, necessitating a careful assessment of the effect of formulation excipients on the permeation as well as the overall therapeutic outcomes. The in vitro transport data from the preclinical investigations, using laboratory animal models and/or in vitro cell culture systems, must be carefully validated in vivo as there are differences between these models and the human skin. Mathematical models such as MTMT that utilize the interdependence of certain physicochemical characteristics and percutaneous permeability have a predictive value in assessing the flux behavior of enantiomers and racemates. Chirality 15:456,465, 2003. © 2003 Wiley-Liss, Inc. [source] Anti-allergic effects of PG102, a water-soluble extract prepared from Actinidia arguta, in a murine ovalbumin-induced asthma modelCLINICAL & EXPERIMENTAL ALLERGY, Issue 2 2009D. Kim Summary Background Asthma is a chronic inflammatory disease of the lung and its incidence has been increasing around the world. We previously reported that oral administration of a water-soluble extract prepared from Actinidia arguta, code-named PG102, could modulate the level of Th1 and Th2 cytokines and suppress the production of immunoglobulin E (IgE) in the ovalbumin (OVA)-immunized murine model as well as in the in vitro cell culture system, and furthermore could significantly improve dermatitis conditions in the NC/Nga murine model. These data suggested that PG102 might have therapeutic effects in a broad range of allergic diseases. Objective To assess the possible anti-allergic effects of PG102 in the OVA-induced murine asthma model. Methods The quality of PG102 was standardized, using its effects on the production of IgE, IL-5, and IL-13, in in vitro cell culture systems. To test effects on asthma, BALB/c mice were orally administrated with PG102, followed by OVA sensitization and challenge to induce asthmatic symptoms. Airway hyperresponsiveness (AHR), bronchoalveolar lavage fluid, serum, and lung tissue were analysed by using various methods. Results PG102 could decrease the level of IgE, IL-5, and IL-13 in in vitro cell culture systems with IC50 being 1.12,1.43 mg/mL. PG102 could ameliorate asthmatic symptoms, including AHR and eosinophilia in the lungs. Such improvement of asthmatic symptoms by PG102 was accompanied by the down-regulation of IL-5 and IgE. In PG102-treated mice, high level expression of heme oxygenase-1, a potent anti-inflammatory enzyme, was observed in alveolar inflammatory cells, while the mRNA levels of foxp3, TGF-,1, and IL-10, important markers for regulatory T cells, were also up-regulated in the lung tissue. Conclusions PG102 may have potential as a safe and effective reagent for the prevention or treatment of asthma. [source] | |||||||||||||