In-vitro test system for the evaluation of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) inhibitors based on a single HPLC run with UV detection using bovine aortic coronary endothelial cells (BAECs)

2001 ◽  
Vol 50 (5) ◽  
pp. 262-269 ◽  
Author(s):  
G. Dannhardt ◽  
H. Ulbrich
Keyword(s):  
Endothelial Cells ◽  
Test System ◽  
In Vitro Test ◽  
Cyclooxygenase 1 ◽  
Vitro Test ◽  
Cox 2 ◽  
Coronary Endothelial Cells ◽  
Cox 2 Inhibitors ◽  
Cox 1

Retraction Of Fibrin Clots By Cultured Endothelial Cells

1981 ◽  
Author(s):  
B Barbieri ◽  
G Balconi ◽  
E Dejana ◽  
M B Donati
Keyword(s):  
Endothelial Cells ◽  
Primary Cultures ◽  
Fibrin Clot ◽  
Test System ◽  
In Vitro Test ◽  
Clot Retraction ◽  
System L ◽  
Vitro Test ◽  
Fibrin Clots

The interactions between endothelial cells (EC) and fibrin are still poorly understood. We approached this problem by studying the ability of cultured EC to induce in vitro the retraction of fibrin clots. EC were obtained from bovine aorta and human umbilical veins by collagenase treatment and grown in Eagle MEM. At the time of the test the cells were harvested from the flask by a short trypsin-EDTA treatment and resuspended in tyrode solution. The test system involved incubation of the cell suspension in a water-bath at 37°C in the presence of cell-free plasma which was clotted by thrombin. The course of retraction was followed by measuring the diameter of the clot with a microcaliper. Retraction values were expressed after calculation of percent activity by an appropriate formula. EC were found to induce the retraction of the fibrin clot to an extent which increased with the time (1-24 h) and with the number of cells in the system (l-4×l06/ml f.c.). Fibrin clot retractile (FCR) activity of EC could not be detected at 22°C or in presence of Na2-EDTA or using mechanically disrupted cells. Moreover, using batroxobin instead of thrombin as a clotting agent, no retraction occurred; FCR of EC thus showed many characteristics in common with platelet- and fibroblast- induced clot retraction.FCR activity of bovine EC increased with the number of subcultures, being very low in cells harvested from primary cultures. In contrast, human EC had high activity in primary cultures. Similarly to fibroblasts, EC with higher density in culture showed lower FCR, suggesting that con-fluency inhibits the cell contractile capacity.FCR could thus represent a simple in vitro test to further characterize the biology of EC and to evaluate their role in the development of fibrin thrombi.

Development of an in-vitro test system for the evaluation of cyclooxygenase-2 inhibitors

1999 ◽  
Vol 48 (3) ◽  
pp. 133-138 ◽  
Author(s):  
S. Laufer ◽  
P. Zechmeister ◽  
T. Klein
Keyword(s):  
Test System ◽  
Cyclooxygenase 2 ◽  
In Vitro Test ◽  
Vitro Test ◽  
Cyclooxygenase 2 Inhibitors

Cyclooxygenase-2 plays a significant role in regulating the tone of the fetal lamb ductus arteriosus

1999 ◽  
Vol 276 (3) ◽  
pp. R913-R921 ◽  
Author(s):  
Ronald I. Clyman ◽  
Pierre Hardy ◽  
Nahid Waleh ◽  
Yao Qi Chen ◽  
Françoise Mauray ◽  
...  
Keyword(s):  
Significant Role ◽  
Ductus Arteriosus ◽  
Late Gestation ◽  
Relative Contribution ◽  
Cox Inhibitor ◽  
Fetal Lamb ◽  
Cox 2 ◽  
Cox 2 Inhibitors ◽  
Cox 1

Nonselective cyclooxygenase (COX) inhibitors are potent tocolytic agents but have adverse effects on the fetal ductus arteriosus. We hypothesized that COX-2 inhibitors may not affect the ductus if the predominant COX isoform is COX-1. To examine this hypothesis, we used ductus arteriosus obtained from late-gestation fetal lambs. In contrast to our hypothesis, fetal lamb ductus arteriosus expressed both COX-1- and COX-2-immunoreactive protein (by Western analysis). Although COX-1 was found in both endothelial and smooth muscle cells, COX-2 was found only in the endothelial cells lining the ductus lumen (by immunohistochemistry). The relative contribution of COX-1 and COX-2 to PGE2 synthesis was consistent with the immunohistochemical results: in the intact ductus, PGE2 formation was catalyzed by both COX-1 and COX-2 in equivalent proportions; in the endothelium-denuded ductus, COX-2 no longer played a significant role in PGE2 synthesis. NS-398, a selective inhibitor of COX-2, was 66% as effective as the selective COX-1 inhibitor valeryl salicylate and the nonselective COX inhibitor indomethacin in causing contraction of the ductus in vitro. At this time, caution should be used when recommending COX-2 inhibitors for use in pregnant women.

Development of an in vitro test system to identify compounds with cell transforming activity

2009 ◽  
Vol 189 ◽  
pp. S67
Author(s):  
René Thierbach ◽  
Urte Blume ◽  
Pablo Steinberg
Keyword(s):  
Test System ◽  
In Vitro Test ◽  
Transforming Activity ◽  
Vitro Test ◽  
Cell Transforming

Involvement of cyclooxygenase 2 (COX-2) in intrinsic tone of isolated guinea pig

1995 ◽  
Vol 73 (11) ◽  
pp. 1561-1567 ◽  
Author(s):  
L. Charette ◽  
C. Misquitta ◽  
J. Guay ◽  
D. Riendeau ◽  
T. R. Jones
Keyword(s):  
Guinea Pig ◽  
Time Course ◽  
Cyclooxygenase 2 ◽  
Maximal Response ◽  
Pharmacological Agents ◽  
Cyclooxygenase 1 ◽  
Inflammatory Drugs ◽  
Cox 2 ◽  
Cox 2 Inhibitors ◽  
Cox 1

Indomethacin and related nonsteroidal anti-inflammatory drugs relax prostanoid-dependent intrinsic tone of isolated guinea pig trachea by inhibiting cyclooxygenase (COX). Recently, a second isoform of COX (COX-2) was discovered, which differed from COX-1 with respect to protein structure, transcriptional regulation, and susceptibility to inhibition by pharmacological agents. It is now known that indomethacin nonselectively inhibits COX-1 and COX-2, whereas NS-398 is a selective inhibitor of COX-2. In the present study we compared the activity of a selective (NS-398) and nonselective (indomethacin) COX-2 inhibitor on intrinsic tone of isolated guinea pig trachea. NS-398 ≥ indomethacin produced a reversal of intrinsic tone with a similar concentration-dependent (10 nM to 1 μM) time course (Tmax approximately 20–45 min), potency (EC50 1.7 and 5.6 nM, respectively), and maximal response. Contractions to cholinergic nerve stimulation (45 V, 0.5 ms, 0.1–32 Hz) and histamine were similarly modulated in tissues relaxed with the selective or nonselective COX-2 inhibitors. Immunoblot analyses showed that COX-2 protein synthesis was induced in both the cartilage and smooth muscle portions of the trachea during changes in intrinsic tone. These findings are consistent with pharmacological results and provide the first demonstration that prostanoid tone in isolated guinea pig trachea is dependent on COX-2 activity. The results also suggest that the activity of indomethacin in this preparation is likely related to COX-2 inhibition.Key words: cyclooxygenase 2, relaxation, guinea pig trachea, cyclooxygenase 1.

An in vitro test system for compounds that modulate human inflammatory macrophage polarization

2018 ◽  
Vol 833 ◽  
pp. 328-338 ◽  
Author(s):  
Hiromi Shiratori ◽  
Carmen Feinweber ◽  
Sonja Luckhardt ◽  
Nadja Wallner ◽  
Gerd Geisslinger ◽  
...  
Keyword(s):  
Macrophage Polarization ◽  
Test System ◽  
In Vitro Test ◽  
Vitro Test ◽  
Inflammatory Macrophage

scholarly journals Evaluation of Anti-Inflammatory Components of Guizhi Fuling Capsule, an Ancient Chinese Herbal Formula, in Human Umbilical Vein Endothelial Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-8 ◽  
Author(s):  
Yuzhong Zheng ◽  
Guizhong Xin ◽  
Guowei Gong ◽  
Tina TX Dong ◽  
Ping Li ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Active Components ◽  
Human Umbilical Vein ◽  
Cellular Inflammation ◽  
Cyclooxygenase 1 ◽  
Cox 2 ◽  
Anti Inflammatory ◽  
Umbilical Vein Endothelial Cells ◽  
Cox 1

Background. Guizhi Fuling capsule (GFC), a well-known formula composed of five medicinal herbs, is commonly prescribed to treat primary dysmenorrhea, as well as to achieve good clinical efficacy in China. However, the active components of GFC have not been identified. Here, the anti-inflammatory functions of GFC, as well as its major ingredients, were evaluated in human umbilical vein endothelial cells (HUVECs). Methods. Lipopolysaccharide (LPS) was used in HUVECs to imitate the cellular inflammation. Then, GFC-triggered mRNA expressions of cyclooxygenase-1 (COX-1) and COX-2 were determined by real-time PCR, while the expression of COX-2 protein was revealed by western blotting. Besides, nine components of GFC were evaluated for their contribution value in the anti-dysmenorrhea effects Results. The application of GFC downregulated the mRNA expressions of COX-1 and COX-2 mRNAs. Nine major components of GFC were tested in the inflammatory system, and three compounds, including paeoniflorin, benzoylpaeoniflorin, and amygdalin, exhibited robust activation in HUVECs. The combination of paeoniflorin, benzoylpaeoniflorin, and amygdalin showed over 80% of the anti-inflammatory activation. Conclusion. Our study supports that GFC plays a promising role in anti-dysmenorrhea function by decreasing COXs’ expression. Besides, paeoniflorin, benzoylpaeoniflorin, and amygdalin could be considered as major regulators for the anti-dysmenorrhea effects of GFC.

Biomimetic in vitro test system for evaluation of dental implant materials

2020 ◽  
Vol 36 (8) ◽  
pp. 1059-1070
Author(s):  
Franziska Ehlicke ◽  
Jonathan Berndt ◽  
Nina Marichikj ◽  
Doris Steinmüller-Nethl ◽  
Heike Walles ◽  
...  
Keyword(s):  
Dental Implant ◽  
Test System ◽  
In Vitro Test ◽  
Implant Materials ◽  
Vitro Test

scholarly journals Prediction of in Vivo Cytotoxicity of Chemotherapeutic Agents by Their Effect on Malignant Leukocytes in Vitro

Blood ◽  
1967 ◽  
Vol 30 (2) ◽  
pp. 176-188 ◽  
Author(s):  
MARTIN J. CLINE
Keyword(s):  
Test System ◽  
Chemotherapeutic Agents ◽  
Response To Treatment ◽  
In Vitro Test ◽  
Malignant Cells ◽  
Vitro Test ◽  
In Vitro Effects ◽  
In Vivo Cytotoxicity

Abstract In order to develop a test system for predicting the response to chemotherapeutic agents, leukocytes from patients with leukemia and leukolymphosarcoma were cultured in vitro and the effect of several drugs on the incorporation of H3-uridine into ribonucleic acid was measured. Cortisol, vincristine and cytosine arabinoside at concentrations near the therapeutic range produced inhibition of H3-uridine incorporation in sensitive leukocytes. The in vitro effects of 6-mercaptopurine and methotrexate were variable. In 39 trials on 25 patients with leukemia or lymphosarcoma, the in vitro test was used successfully to predict the response to treatment with prednisone and vincristine. It was concluded that the in vitro test system can predict the in vivo cytotoxicity of certain drugs for malignant cells, although it cannot be used to predict the likelihood of the induction of remissions with these drugs.

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