scholarly journals Effect of nitric oxide release from NOR-3 on urea synthesis, viability and oxygen consumption of rat hepatocyte cultures

2007 ◽  
pp. 427-432
Author(s):  
R Chimenti ◽  
G Martino ◽  
S Mazzulla ◽  
S Sesti
Keyword(s):  
Nitric Oxide ◽  
Oxygen Consumption ◽  
Cell Viability ◽  
Urea Synthesis ◽  
Rat Hepatocyte ◽  
Trypan Blue Exclusion ◽  
No Donor ◽  
Nitric Oxide Release ◽  
Hepatocyte Cultures ◽  
Trypan Blue Exclusion Test

As nitric oxide is considered a mediator of liver oxidative metabolism during sepsis, we studied the effects of exogenous nitric oxide, produced by NO-donor, (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (NOR-3), on cell viability, urea biosynthesis and oxygen consumption in rat hepatocyte cultures. Nitric oxide release from NOR-3 was studied using 4,5-diaminofluorescein diacetate. Urea levels were measured by the spectrophotometric method. Cell viability was determined by the MTT test and trypan blue exclusion test, whereas oxygen consumption was measured by a polarographic technique. After 2 h treatment, NOR-3 induced an increase in the levels of nitric oxide. After 2 h of treatment and 24 h after the end of the treatment with NOR-3, both cell viability and urea synthesis were significantly reduced in comparison to the controls for NOR-3 concentrations equal to or greater than 50 microM. A reduction in oxygen consumption was observed in hepatocytes after 40 min treatment with 100 microM NOR-3, even if the cell viability was unchanged. Reduction of oxygen consumption is an early indicator of the metabolic alterations in hepatocytes exposed to nitric oxide. These findings suggest that nitric oxide accumulation acts on hepatocyte cultures inducing cell death and reduction of urea synthesis after 2 hours.

scholarly journals Suitability of a Progenitor Cell-Enriching Device for In Vitro Applications

Coatings ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 146
Author(s):  
Antonio Celentano ◽  
Tami Yap ◽  
Giuseppe Pantaleo ◽  
Rita Paolini ◽  
Michael McCullough ◽  
...  
Keyword(s):  
Cell Viability ◽  
Time Course ◽  
Ex Vivo ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Test ◽  
Initial Reduction ◽  
Metal Wear ◽  
Class 1 ◽  
Electron Energy Loss

Rigenera® is a novel class-1 medical device that produces micro-grafts enriched of progenitors cells without ex vivo manipulation of donor tissues. The manufacturer’s protocol has been supported for a wide variety of clinical uses in the field of regenerative medicine. This study aimed to evaluate its potential use for in vitro cell models. Human primary oral fibroblasts were cultured under standard conditions and processed through Rigenera® over a time course of up to 5 min. Cell viability was assessed using a Trypan Blue exclusion test. It is possible to process fibroblasts through Rigenera® although an initial reduction of cell viability was observed. Additionally, debris was evident in the cell suspension of the processed samples. Scanning electron microscopy (SEM) microanalysis of the debris and electron energy-loss spectroscopy confirmed the presence of metal wear possibly due to the processing conditions used in this study. Interestingly, pore sizes within Rigeneracons® grids were found to range between 250–400 μm. This is the first report assessing the suitability of Rigenera® and Rigeneracons® for in vitro applications. Whilst Rigenera® workflow was found to be amenable to laboratory uses, our results strongly suggest that further research and development is necessary to support the utilization of this technology for enrichment of micro-graft derived cells and cell sorting in vitro.

Nitric oxide release from a cucurbituril encapsulated NO-donor

2018 ◽  
Vol 16 (23) ◽  
pp. 4272-4278 ◽  
Author(s):  
A. Acuña ◽  
N. Basílio ◽  
M. Parajó ◽  
J. C. Mejuto ◽  
J. Pérez-Juste ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Chemical Stimulus ◽  
No Donor ◽  
Nitric Oxide Release

The denitrosation of a S-nitrosothiol derivative, nitrosomercaptopyridine (SNO+), can be inhibited by incorporation into the cucurbit[7]uril cavity. Owing to the reversible character of host : guest complexation, SNO+ can be expelled from the host cavity through the application of a chemical stimulus allowing controlled nitric oxide release.

scholarly journals Dental Pulp Fibroblasts Response after Stimulation with HEMA and Adhesive System

2018 ◽  
Vol 29 (5) ◽  
pp. 419-426 ◽  
Author(s):  
Karin Cristina da Silva Modena ◽  
Adriana Maria Calvo ◽  
Carla Renata Sipert ◽  
Thiago José Dionísio ◽  
Maria Fidela de Lima Navarro ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nitric Oxide ◽  
Dental Pulp ◽  
Close Contact ◽  
Adhesive System ◽  
Single Bond ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method ◽  
Nitric Oxide Release

Abstract This study evaluated in vitro cell viability and metabolism, nitric oxide release and production of chemokines by cultured human dental pulp fibroblasts (DPF) under contact with HEMA and Single Bond. Cultures of DPF were established by means of an explant technique. Once plated, cells were kept under contact with increasing concentrations of HEMA (10, 100 and 1000 nM) or Single Bond (SB) [10-fold serially diluted in culture medium (10-4, 10-3 and 10-2 v/v)] and also with polymerized SB components. Cytotoxicity was assessed by Trypan Blue exclusion method and MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Nitric oxide release on cell supernatant was detected by Griess Method whereas chemokines (CXCL12 and CXCL8) were detected by ELISA. RT-qPCR was employed for chemokines gene expression analysis. Cytotoxic tests showed significant differences for SB 10-2. None of the tested materials significantly altered NO levels. Protein levels of CXCL12 were significantly decreased only by HEMA. On the other hand, while CXCL12 mRNA remained unaltered, gene expression of CXCL8 had significant decrease with all materials, except for polymerized SB. In conclusion, Single Bond and HEMA at various concentrations, decreased expression and production of molecules involved in inflammatory processes and, therefore, the use of adhesive systems such as pulp capping materials must be viewed with caution due to its large cytotoxic effect when in close contact with the pulp.

scholarly journals Glucose release as a response to glucagon in rat hepatocyte culture: involvement of NO signaling

2008 ◽  
pp. 569-575
Author(s):  
H Farghali ◽  
J Hodis ◽  
N Kutinová-Canová ◽  
P Potměšil ◽  
E Kmoníčková ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Glycogen Synthase ◽  
Inos Mrna ◽  
No Production ◽  
Rat Hepatocyte ◽  
No Donor ◽  
Glucagon Receptor ◽  
Nos Inhibitors ◽  
No Signaling ◽  
Rat Hepatocyte Culture

Glucagon and α-adrenergic-induced glycogenolysis is realized via the agonist/adenylyl cyclase/cAMP/protein kinase signaling pathway or via the activation of phosphorylase kinase by the mobilized calcium that supports the inhibition of glycogen synthase, respectively. The role of nitric oxide (NO) in this process has not been extensively studied. The present work was directed to the question whether NO is produced during glucagon-induced glycogenolysis in rat hepatocyte in a similar way like α-adrenoceptor stimulation. Glycogen-rich hepatocyte cultures were used. NO production (NO2-) was assessed under the influence of glucagon, dibutyryl cyclic AMP (db-cAMP), forskolin, the nitric oxide synthase (NOS) inhibitors Nω-nitro-Larginine methyl ester (L-NAME) and aminoguanidine, and the NO donor S-nitroso-N-acetyl penicillamine (SNAP). Inducible NOS (iNOS) mRNA was examined by reverse transcription-polymerase chain reaction. Glycogenolysis was followed up by estimation of medium glucose levels. The amount of glucose and NO2 - released by glycogen-rich hepatocytes was increased as a result of glucagon, db-cAMP, forskolin and SNAP treatments. iNOS gene expression was upregulated by glucagon. Glycogenolysis that occurs through glucagon receptor stimulation involves NO production downstream of transduction pathways through an isoform of NO synthase. The present and previous studies document possible involvement of NO signaling in glycogenolytic response to glucagon and adrenergic agonists in hepatocytes.

Macrophage-induced inhibition of nitric oxide production in primary rat hepatocyte cultures via prostaglandin E2 release

Hepatology ◽  
1998 ◽  
Vol 28 (5) ◽  
pp. 1300-1308 ◽  
Author(s):  
B�n�dicte Griffon ◽  
Josiane Cillard ◽  
Martine Chevanne ◽  
Isabelle Morel ◽  
Pierre Cillard ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Prostaglandin E2 ◽  
Nitric Oxide Production ◽  
Rat Hepatocyte ◽  
Hepatocyte Cultures ◽  
Oxide Production

scholarly journals S-Nitroso-N-Acetyl-D-Penicillamine Modified Hyperbranched Polyamidoamine for High-Capacity Nitric Oxide Storage and Release

2020 ◽  
Vol 7 (1) ◽  
pp. 9
Author(s):  
Sean P. Hopkins ◽  
Megan C. Frost
Keyword(s):  
Nitric Oxide ◽  
High Capacity ◽  
Implantable Devices ◽  
Resonance Spectroscopy ◽  
No Donor ◽  
Ionization Time ◽  
Nitric Oxide Release ◽  
Beneficial Effects ◽  
1H Nuclear Magnetic Resonance

Synthetic nitric oxide (NO)-donating materials have been shown to have many beneficial effects when incorporated into biomedical materials. When released in the correct dosage, NO has been shown to increase the biocompatibility of blood and tissue contacting materials, but materials are often limited in the amount of NO that can be administered over a period of time. To address this, hyperbranched polyamidoamine (HPAMAM) was modified with the S-nitrosothiol, S-nitroso-N-acetyl-D-penicillamine, and nitrosated to form a controlled, high-capacity NO-donating compound (SNAP-HPAMAM). This compound has the potential of modifying polymers to release NO over long periods of time by being blended into a variety of base polymers. Nitric oxide release was triggered by photoinitiation and through passive ion-mediated release seen under physiological conditions. A material that delivers the beneficial dose of NO over a long period of time would be able to greatly increase the biocompatibility of long-term implantable devices. Structural analysis of a generation 2 HPAMAM molecule was done through Fourier transform infrared spectroscopy (FTIR), 1H nuclear magnetic resonance spectroscopy (NMR), and matrix assisted laser desorption ionization, time of flight (MALDI-TOF) mass spectrometry. The NO capacity of the finalized generation 2 SNAP-HPAMAM compound was approximately 1.90 ± 0.116 µmol NO/mg. Quantification of the functional groups in the compound proved that an average of 6.40 ± 0.309 reactive primary amine sites were present compared to the 8 reactive sites on a perfectly synthesized generation 2 dendrimer. There is a substantial advantage of using the hyper-branched HPAMAM over purified dendrimers in terms of reduced labor and expense while still providing a high-capacity NO donor that can be blended into different polymer matrices.

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