Development of an in vitro skin sensitization test based on ROS production in THP-1 cells

Herein, spinal fixation implants were constructed using degradable polymeric materials such as PGA–PLA block copolymers (poly(glycolic acid-b-lactic acid)). These materials were reinforced by blending with HA-g-PLA (hydroxyapatite-graft-poly lactic acid) and PGA fiber before being tested to confirm its biocompatibility via in vitro (MTT assay) and in vivo animal experiments (i.e., skin sensitization, intradermal intracutaneous reaction, and in vivo degradation tests). Every specimen exhibited suitable biocompatibility and biodegradability for use as resorbable spinal fixation materials.

Download Full-text

Vitamin E-Coated Dialyzer Inhibits Oxidative Stress

Blood Purification ◽

10.1159/000478971 ◽

2017 ◽

Vol 44 (4) ◽

pp. 288-293 ◽

Cited By ~ 4

Author(s):

Shiho Yamadera ◽

Yuya Nakamura ◽

Masahiro Inagaki ◽

Isao Ohsawa ◽

Hiromichi Gotoh ◽

...

Keyword(s):

Oxidative Stress ◽

Vitamin E ◽

Synthetic Polymer ◽

Intracellular Reactive Oxygen Species ◽

Ros Production ◽

Oxygen Species ◽

In Vitro Methods ◽

Stress Effects ◽

Intracellular Ros

Aim: To examine the effects of vitamin E-coated dialyzer on oxidative stress in vitro. Methods: A dialyzer with a synthetic polymer membrane (APS-11SA) and vitamin E-coated dialyzer (VPS-11SA) were connected to a blood tubing line, and U937 cells were circulated in the device. The circulating fluid was collected at 1, 2, 5, 10, 25, and 50 cycles, which are estimated numbers of passes through the dialyzer. Intracellular reactive oxygen species (ROS) production, malondialdehyde (MDA), and Cu/Zn-superoxide dismutase (SOD) were quantified. Results: Intracellular ROS production was increased in the first cycle by APS-11SA and was decreased throughout the experiment by VPS-11SA. Intracellular ROS production in the VPS-11SA device was lower, and MDA levels were decreased. MDA levels were lower during VPS-11SA processing than during APS-11SA processing. Cu/Zn-SOD levels remained unchanged. Conclusion: Our results highlight anti-oxidative-stress effects of a vitamin E-coated dialyzer.

Download Full-text

In Vitro Anti-Inflammatory and Immunomodulatory Activities of an Extract from the Roots of Bupleurum rotundifolium

Medicines ◽

10.3390/medicines6040101 ◽

2019 ◽

Vol 6 (4) ◽

pp. 101 ◽

Cited By ~ 2

Author(s):

Cholet ◽

Decombat ◽

Vareille-Delarbre ◽

Gainche ◽

Berry ◽

...

Keyword(s):

Mononuclear Cells ◽

Polymorphonuclear Neutrophils ◽

Ros Production ◽

Peripheral Blood Mononuclear ◽

Aerial Parts ◽

Bupleurum Scorzonerifolium Willd ◽

Anti Inflammatory ◽

Inflammatory Profile ◽

Bupleurum Scorzonerifolium

Background: Some Bupleurum species, such as the Bupleurum chinense DC. or the Bupleurum scorzonerifolium Willd have been extensively studied (especially their roots) for the treatment of inflammation. In contrast, only compounds extracted from the aerial parts of Bupleurum rotundifolium have been studied and showed anti-inflammatory or antiproliferative activities. This study was conducted to investigate the antioxidant, anti-inflammatory, and immunomodulatory effects of Bupleurum rotundifolium roots. Methods: To tackle the various aspects of inflammation, we studied in vitro a methanolic extract from the roots of Bupleurum rotundifolium on peripheral blood mononuclear cells (PBMCs), polymorphonuclear neutrophils (PMNs), and the monocytic cells THP-1. Its antioxidant capacities and iron-chelating activity were assessed. The extract was tested on THP-1 differentiation, reactive oxygen species (ROS) production by leukocytes, neutrophils chemotaxis, cytokines, PGE2 production, and NF-κB activation in PBMCs. Results: The extract showed a decreased ROS production in stimulated cells. It increased PBMC chemokine secretion and up-regulated the differentiation of THP-1 monocytes into macrophage-like cells, indicating a potential interest of the extract in the resolution of acute inflammation. In addition, the analysis of cytokine production suggests that Bupleurum rotundifolium has immunomodulatory properties. Conclusions: Cytokines secretion, especially IL-1β and IL-12p70, provided us with a set of indicators suggesting that the extract might be able to drive the polarization of macrophages and lymphocytes toward a Th2 anti-inflammatory profile in excessive inflammation.

Download Full-text

Hyperglycemia Induces Generation of Reactive Oxygen Species and Accelerates Apoptotic Cell Death in Salivary Gland Cells

Pathobiology ◽

10.1159/000512639 ◽

2021 ◽

pp. 1-8

Author(s):

Naoyuki Matsumoto ◽

Daisuke Omagari ◽

Ryoko Ushikoshi-Nakayama ◽

Tomoe Yamazaki ◽

Hiroko Inoue ◽

...

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Salivary Gland ◽

Tissue Injury ◽

Ros Production ◽

Oxygen Species ◽

Saliva Secretion ◽

Salivary Gland Tissue ◽

Gland Tissue

Introduction: Type-2 diabetes mellitus (T2DM) is associated with several systemic vascular symptoms and xerostomia. It is considered that hyperglycemia-induced polyuria and dehydration cause decreased body-water volume, leading to decreased saliva secretion and, ultimately, xerostomia. In T2DM, increased production of reactive oxygen species (ROS) causes tissue damage to vascular endothelial cells as well as epithelial tissue, including pancreas and cornea. Hence, a similar phenomenon may occur in other tissues and glands in a hyperglycemic environment. Methods: Salivary gland tissue injury was examined, using T2DM model mouse (db/db). Transferase‐mediated dUTP nick‐end labeling (TUNEL) was conducted to evaluate tissue injury. The levels of malondialdehyde (MDA) and 8-hydroxy-2′-deoxyguanosine, Bax/Bcl-2 ratio were measured as indicator of oxidative stress. Moreover, in vitro ROS production and cell injury was evaluated by mouse salivary gland-derived normal cells under high-glucose condition culture. Results: In vivo and in vitro analysis showed a higher percentage of TUNEL-positive cells and higher levels of MDA and 8-hydroxy-2′-deoxyguanosine in salivary gland tissue of db/db mice. This suggests damage of saliva secretion-associated lipids and DNA by hyperglycemic-induced oxidative stress. To analyze the mechanism by which hyperglycemia promotes ROS production, mouse salivary gland-derived cells were isolated. The cell culture with high-glucose medium enhanced ROS production and promotes apoptotic and necrotic cell death. Conclusion: These findings suggest a novel mechanism whereby hyperglycemic-induced ROS production promotes salivary gland injury, resulting in hyposalivation.

Download Full-text

8-Formylophiopogonanone B induces ROS-mediated apoptosis in nasopharyngeal carcinoma CNE-1 cells

Toxicology Research ◽

10.1093/toxres/tfab087 ◽

2021 ◽

Author(s):

Ya-jing Zhang ◽

Zhen-lin Mu ◽

Ping Deng ◽

Yi-dan Liang ◽

Li-chuan Wu ◽

...

Keyword(s):

Nasopharyngeal Carcinoma ◽

Cell Viability ◽

Tumor Cells ◽

High Efficiency ◽

Pharmacological Action ◽

Migration And Invasion ◽

Ros Production ◽

Biological Functions ◽

Intracellular Ros

Abstract Cancer is one of the leading causes of death in the world. It is very important to find drugs with high efficiency, low toxicity, and low side effects for the treatment of cancer. Flavonoids and their derivatives with broad biological functions have been recognized as anti-tumor chemicals. 8-Formylophiopogonanone B (8-FOB), a naturally existed homoisoflavonoids with rarely known biological functions, needs pharmacological evaluation. In order to explore the possible anti-tumor action of 8-FOB, we used six types of tumor cells to evaluate in vitro effects of this agent on cell viability and tested the effects on clone formation ability, scratching wound-healing, and apoptosis. In an attempt to elucidate the mechanism of pharmacological action, we examined 8-FOB-induced intracellular oxidative stress and -disrupted mitochondrial function. Results suggested that 8-FOB could suppress tumor cell viability, inhibit cell migration and invasion, induce apoptosis, and elicit intracellular ROS production. Among these six types of tumor cells, the nasopharyngeal carcinoma CNE-1 cells were the most sensitive cancer cells to 8-FOB treatment. Intracellular ROS production played a pivotal role in the anti-tumor action of 8-FOB. Our present study is the first to document that 8-FOB has anti-tumor activity in vitro and increases intracellular ROS production, which might be responsible for its anti-tumor action. The anti-tumor pharmacological effect of 8-FOB is worthy of further investigation.

Download Full-text

Evaluation of the Skin Sensitization Potential of Silica nanoparticles using in vitro and in vivo assay

Toxicology Letters ◽

10.1016/s0378-4274(21)00415-x ◽

2021 ◽

Vol 350 ◽

pp. S72

Author(s):

D.H. Lee ◽

S.-H. Kim ◽

J.H. Lee ◽

J.-Y Yang ◽

H.-S. Shin ◽

...

Keyword(s):

Silica Nanoparticles ◽

Skin Sensitization ◽

In Vivo Assay

Download Full-text

The relationship between CD86/CD54 expression and THP-1 cell viability in an in vitro skin sensitization test – human cell line activation test (h-CLAT)

Cell Biology and Toxicology ◽

10.1007/s10565-008-9059-9 ◽

2008 ◽

Vol 25 (2) ◽

pp. 109-126 ◽

Cited By ~ 62

Author(s):

Hitoshi Sakaguchi ◽

Takao Ashikaga ◽

Masaaki Miyazawa ◽

Nanae Kosaka ◽

Yuichi Ito ◽

...

Keyword(s):

Cell Viability ◽

Cell Line ◽

Human Cell ◽

Human Cell Line ◽

Skin Sensitization ◽

Activation Test ◽

The Relationship

Download Full-text

Callicarpa Nudiflora Extract Exerts Anti-Inflammatory Effect on H. Pylori-Associated Gastritis Through Repression of ROS/NLRP3/Caspase-1/IL-1β Signaling Axis

10.21203/rs.3.rs-588224/v1 ◽

2021 ◽

Author(s):

Lili Li ◽

Xiaohui Zhu ◽

Xingxing Chai ◽

Xiaoyu Chen ◽

Xiaohua Su ◽

...

Keyword(s):

Cell Damage ◽

Underlying Mechanism ◽

Inflammatory Factors ◽

Inflammasome Activation ◽

Ros Production ◽

Gastric Diseases ◽

Caspase 1 ◽

Signaling Axis ◽

H Pylori

Abstract Helicobacter pylori ( H. pylori ) is a major pathogenic factor for the development of gastric diseases including chronic gastritis and gastric cancer. Callicarpa nudiflora (CN), an air-dried leaves extract of Callicarpa nudiflora Hook. & Arn., has been found to exhibit a broad-spectrum antibacterial effect. In our study, we extracted the active ingredient from air-dried leaves of Callicarpa nudiflora, detected the effect of CN against H. pylori -infected GES-1 cells in vitro , and elucidated the underlying mechanism. GES-1 cells were cocultured with HPSS1 at MOI = 100:1 and treated with different concentrations of CN. Results indicated that CN not only significantly decreased cellular lactate dehydrogenase leakage, but also markedly attenuated H. pylori -induced cell apoptosis and ROS production in GSE-1 cells, therefore protecting gastric epithelial cells against injuries caused by H. pylori . CN also inhibited the secretions of inflammatory factors, such as tumor necrosis factor-α (TNF-α), IL-1β, IL-6 and IL-8. Furthermore, CN remarkably decreased the expression levels of NLRP3, PYCARD, active Caspase-1. In conclusion, CN exhibited highly efficient protective effect against H. pylori -induced gastritis and cell damage; Mechanismly, CN suppressed H. pylori -triggered inflammatory response and pyroptosis through depressing ROS production and NLRP3 inflammasome activation via ROS/NLRP3/IL-1β signaling axis.

Download Full-text

Protection of the vascular endothelium in experimental situations

Interdisciplinary Toxicology ◽

10.2478/v10102-011-0005-y ◽

2011 ◽

Vol 4 (1) ◽

pp. 20-26 ◽

Cited By ~ 16

Author(s):

Ružena Sotníková ◽

Jana Nedelčevová ◽

Jana Navarová ◽

Viera Nosáľová ◽

Katarína Drábiková ◽

...

Keyword(s):

Oxidative Stress ◽

Vascular Endothelium ◽

Vascular Dysfunction ◽

Antioxidant Properties ◽

Pharmacological Intervention ◽

Ros Production ◽

Nitric Oxide Radical ◽

Endothelium Dependent Relaxation

Protection of the vascular endothelium in experimental situationsOne of the factors proposed as mediators of vascular dysfunction observed in diabetes is the increased generation of reactive oxygen species (ROS). This provides support for the use of antioxidants as early and appropriate pharmacological intervention in the development of late diabetic complications. In streptozotocin (STZ)-induced diabetes in rats we observed endothelial dysfuction manifested by reduced endothelium-dependent response to acetylcholine of the superior mesenteric artery (SMA) and aorta, as well as by increased endothelaemia. Changes in endothelium-dependent relaxation of SMA were induced by injury of the nitric oxide radical (·NO)-signalling pathway since the endothelium-derived hyperpolarising factor (EDHF)-component of relaxation was not impaired by diabetes. The endothelial dysfunction was accompanied by decreased ·NO bioavailabity as a consequence of reduced activity of eNOS rather than its reduced expression. The results obtained using the chemiluminiscence method (CL) argue for increased oxidative stress and increased ROS production. The enzyme NAD(P)H-oxidase problably participates in ROS production in the later phases of diabetes. Oxidative stress was also connected with decreased levels of reduced glutathione (GSH) in the early phase of diabetes. After 10 weeks of diabetes, adaptational mechanisms probably took place because GSH levels were not changed compared to controls. Antioxidant properties of SMe1EC2 foundin vitrowere partly confirmedin vivo.Administration of SMe1EC2 protected endothelial function. It significantly decreased endothelaemia of diabetic rats and improved endothelium-dependent relaxation of arteries, slightly decreased ROS-production and increased bioavailability of ·NO in the aorta. Further studies with higher doses of SMe1EC2 may clarify the mechanism of its endothelium-protective effectin vivo.

Download Full-text

Protective effect of nigella sativa extract and thymoquinone on serum/glucose deprivation-induced PC12 cells death

European Psychiatry ◽

10.1016/s0924-9338(11)72613-x ◽

2011 ◽

Vol 26 (S2) ◽

pp. 908-908

Author(s):

H.R. Sadeghnia ◽

S.H. Mousavi ◽

Z. Tayarani-Najaran ◽

M. Asghari

Keyword(s):

Cell Viability ◽

Pc12 Cells ◽

Neurodegenerative Disorders ◽

Nigella Sativa ◽

Glucose Deprivation ◽

Serum Glucose ◽

Ros Production ◽

Protective Effects ◽

Intracellular Ros

The serum/glucose deprivation (SGD)-induced cell death in cultured PC12 cells represents a useful in vitro model for the study of brain ischemia and neurodegenerative disorders.Nigella sativa L. and its active component, thymoquinone (TQ) have been known as a source of antioxidants. In the present study, the protective effects of N. sativa and TQ on cell viability and reactive oxygen species (ROS) production in cultured PC12 cells were investigated under SGD conditions. PC12 Cells were pretreated with different concentrations of N. sativa extract (15.62–250 μg/ml) and TQ (1.17–150 μM) for 2 h and then subjected to SGD for 6 or 18 h. Cell viability was quantitated by MTT assay. Intracellular ROS production was measured by flow cytometry using 2’,7’-dichlorofluorescin diacetate (DCF-DA) as a probe. SGD induced significant cells toxicity after 6, 18, or 24 h (p < 0.001). Pretreatment with N. sativa (15.62–250 μg/ml) and TQ (1.17–37.5 μM) reduced SGD-induced cytotoxicity in PC12 cells after 6 and 18 h. A significant increase in intracellular ROS production was seen following SGD (p < 0.001). N. sativa (250 μg/ml, p < 0.01) and TQ (2.34, 4.68, 9.37 μM, p < 0.01) pretreatment reversed the increased ROS production following ischemic insult. The experimental results suggest that N. sativa extract and TQ protects the PC12 cells against SGD-induced cytotoxicity via antioxidant mechanisms. Our findings might raise the possibility of potential therapeutic application of N. sativa extract and TQ for managing cerebral ischemic and neurodegenerative disorders.

Download Full-text