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Ex vivo culture of human atherosclerotic plaque culture

Anna Lebedeva, Daria Vorobyeva, Murad Vagida, Oxana Ivanova, Eugeny Felker, Wendy Fitzgerald, Natalya Danilova, Vladimir Gontarenko, Alexander Shpektor, Elena Vasilieva, Leonid Margolis
BACKGROUND AND AIMS: The mechanisms that drive atherosclerotic plaque progression and destabilization in humans remain largely unknown. Laboratory models are needed to study these mechanisms under controlled conditions. The aim of this study was to establish a new ex vivo model of human atherosclerotic plaques that preserves the main cell types in plaques and the extracellular components in the context of native cytoarchitecture. METHODS: Atherosclerotic plaques from carotid arteries of 28 patients undergoing carotid endarterectomy were dissected and cultured...
December 2017: Atherosclerosis
Xiaofeng Yang, Jingyuan Wei, Yanhao He, Ting Jing, Yanxiang Li, Yunfang Xiao, Bo Wang, Weirong Wang, Jiye Zhang, Rong Lin
SIRT1, a highly conserved NAD(+)-dependent protein deacetylase, plays a pivotal role in the pathogenesis and therapy of atherosclerosis (AS). The aim of this study is to investigate the potential effects of SIRT1 on AS in ApoE(-/-) mice and the underlying mechanisms of autophagy in an ox-LDL-stimulated human monocyte cell line, THP-1. In vivo, the accelerated atherosclerotic progression of mice was established by carotid collar placement; then, mice were treated for 4 weeks with a SIRT1-specific inhibitor, EX-527...
August 1, 2017: Oncotarget
Maria Lombardi, Maria Elena Mantione, Domenico Baccellieri, David Ferrara, Renata Castellano, Roberto Chiesa, Ottavio Alfieri, Chiara Foglieni
In atherosclerosis, matrix metallopeptidases (MMPs) contribute to plaque rupture through weakening of the fibrous cap. Pleiotropic P2X purinoceptor 7 (P2X7), expressed in the carotid plaque (PL), is involved in interleukin 1 beta (IL-1β) release that may influence MMP9 generation, thus their possible modulation through acting on P2X7 was investigated. P2X7-related machinery was characterized and the effects of P2X7 antagonists (A740003, KN62) and MMPs inhibitors (Batimastat, Ro28-2653) were studied in ex-vivo tissue cultures of human PL's vs...
July 7, 2017: Scientific Reports
Xiaofeng Yang, Jingyuan Wei, Yanhao He, Ting Jing, Yanxiang Li, Yunfang Xiao, Bo Wang, Weirong Wang, Jiye Zhang, Rong Lin
SIRT1, a highly conserved NAD+-dependent protein deacetylase, plays a pivotal role in the pathogenesis and therapy of atherosclerosis (AS). The aim of this study is to investigate the potential effects of SIRT1 on AS in ApoE-/- mice and the underlying mechanisms of autophagy in an ox-LDL-stimulated human monocyte cell line, THP-1. In vivo, the accelerated atherosclerotic progression of mice was established by carotid collar placement; then, mice were treated for 4 weeks with a SIRT1-specific inhibitor, EX-527...
May 8, 2017: Oncotarget
D A Vorobyova, A M Lebedev, M S Vagida, O I Ivanova, E I Felker, V N Gontarenko, A V Shpektor, L B Margolis, E Yu Vasilieva
THE AIM OF THE STUDY: to analyze the dynamics of lymphocytic composition of human atherosclerotic plaques in ex vivo culture system. MATERIALS AND METHODS: The study included 15 atherosclerotic plaques obtained from patients who underwent carotid endarterectomy. Plaques were cultured as ring-shaped explants on collagen rafts in culture medium of special composition in CO2 incubator according to the previously developed technique. On day 0, and also on the 4th and 19th days of culture we extracted cells from plaque explants and analyzed B- and T-lymphocytic content of the tissue, as well as the percentage of CD16+ natural killer (NK) cells, using multichromatic flow cytometry...
December 2016: Kardiologiia
Glykeria Karadimou, Lasse Folkersen, Martin Berg, Ljubica Perisic, Andrea Discacciati, Joy Roy, Göran K Hansson, Jonas Persson, Gabrielle Paulsson-Berne
AIMS: Processes in the development of atherosclerotic lesions can lead to plaque rupture or erosion, which can in turn elicit myocardial infarction or ischaemic stroke. The aims of this study were to determine whether Toll-like receptor 7 (TLR7) gene expression levels influence patient outcome and to explore the mechanisms linked to TLR7 expression in atherosclerosis. METHODS AND RESULTS: Atherosclerotic plaques were removed by carotid endarterectomy (CEA) and subjected to gene array expression analysis (n = 123)...
January 2017: Cardiovascular Research
Caitlin O'Rourke, Georgia Shelton, Joshua D Hutcheson, Megan F Burke, Trejeeve Martyn, Timothy E Thayer, Hannah R Shakartzi, Mary D Buswell, Robert E Tainsh, Binglan Yu, Aranya Bagchi, David K Rhee, Connie Wu, Matthias Derwall, Emmanuel S Buys, Paul B Yu, Kenneth D Bloch, Elena Aikawa, Donald B Bloch, Rajeev Malhotra
Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis...
May 31, 2016: Journal of Visualized Experiments: JoVE
W Eilenberg, S Stojkovic, A Piechota-Polanczyk, C Kaun, S Rauscher, M Gröger, M Klinger, J Wojta, C Neumayer, I Huk, S Demyanets
OBJECTIVE: Neutrophil gelatinase-associated lipocalin (NGAL), a protein found in activated neutrophils, is expressed in kidney tubule cells in response to noxious stimuli, and is thus recognized as a marker of acute kidney injury. Recent studies have suggested that NGAL could also have pathophysiological importance in cardiovascular diseases. The aim of the present study was to examine NGAL expression in human carotid endarterectomy tissues ex vivo as well as the effects of NGAL in the main cell types involved in atherogenesis, namely in human macrophages, endothelial cells, and smooth muscle cells in vitro...
May 2016: European Journal of Vascular and Endovascular Surgery
Daniel R Lewis, Latrisha K Petersen, Adam W York, Sonali Ahuja, Hoonbyung Chae, Laurie B Joseph, Saum Rahimi, Kathryn E Uhrich, Paul B Haser, Prabhas V Moghe
AIMS: Atherosclerotic development is exacerbated by two coupled pathophysiological phenomena in plaque-resident cells: modified lipid trafficking and inflammation. To address this therapeutic challenge, we designed and investigated the efficacy in vitro and ex vivo of a novel 'composite' nanotherapeutic formulation with dual activity, wherein the nanoparticle core comprises the antioxidant α-tocopherol and the shell is based on sugar-derived amphiphilic polymers that exhibit scavenger receptor binding and counteract atherogenesis...
February 1, 2016: Cardiovascular Research
Shigeko Arita-Okubo, Joo-Ri Kim-Kaneyama, Xiao-Feng Lei, Wen-Guang Fu, Koji Ohnishi, Motohiro Takeya, Aya Miyauchi, Hirokazu Honda, Hiroyuki Itabe, Takuro Miyazaki, Akira Miyazaki
AIMS: The adhesion of circulating monocytes to endothelial cells (ECs) is an early and critical event in the formation of atherosclerotic plaques. Hydrogen peroxide-inducible clone 5 (Hic-5) serves as an adaptor molecule in cell adhesion complexes. However, the role of endothelial Hic-5 in monocyte-EC interaction and atherogenesis remains unclear. We examined the roles of endothelial Hic-5 in monocyte-EC interaction and atherogenesis using mouse models of atherosclerosis and cultured human umbilical vein endothelial cells (HUVECs)...
March 1, 2015: Cardiovascular Research
Christian Erbel, Deniz Okuyucu, Mohammadreza Akhavanpoor, Li Zhao, Susanne Wangler, Maani Hakimi, Andreas Doesch, Thomas J Dengler, Hugo A Katus, Christian A Gleissner
Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic lesions. Mouse models are an important tool to investigate inflammatory processes in atherogenesis, but these models suffer from the phenotypic and functional differences between the murine and human immune system. In vitro cell experiments are used to specifically evaluate cell type-dependent changes caused by a substance of interest, but culture-dependent variations and the inability to analyze the influence of specific molecules in the context of the inflammatory compound in atherosclerotic lesions limit the impact of the results...
2014: Journal of Visualized Experiments: JoVE
Sigrun Badrnya, Waltraud C Schrottmaier, Julia B Kral, Koon-Chu Yaiw, Ivo Volf, Gernot Schabbauer, Cecilia Söderberg-Nauclér, Alice Assinger
OBJECTIVE: A growing body of evidence indicates that platelets contribute to the onset and progression of atherosclerosis by modulating immune responses. We aimed to elucidate the effects of oxidized low-density lipoprotein (OxLDL) on platelet-monocyte interactions and the consequences of these interactions on platelet phagocytosis, chemokine release, monocyte extravasation, and foam cell formation. APPROACH AND RESULTS: Confocal microscopy and flow cytometric analysis revealed that in vitro and in vivo stimulation with OxLDL resulted in rapid formation of platelet-monocyte aggregates, with a preference for CD16+ monocyte subsets...
March 2014: Arteriosclerosis, Thrombosis, and Vascular Biology
Eugene O Apostolov, Ercan Ok, Samuel Burns, Safia Nawaz, Alena Savenka, Sudhir V Shah, Alexei G Basnakian
AIM: Both oxidized LDL and carbamylated LDL are considered important for initiating atherosclerosis in patients with end-stage kidney disease through vascular endothelial cell dysfunction or injury. However their effects on each other and their relationship related to pro-atherosclerotic effects on endothelial cells and macrophages have not been investigated. In this study, we analyzed the competition between LDL carbamylation and oxidation, tested biological effects of carbamylated-oxidized LDL (coxLDL) toward the endothelial cells, assessed its ability to cause foam cell development, and determined the roles of scavenger receptors in this process...
2013: Journal of Atherosclerosis and Thrombosis
Hui-Qing Liu, Xiao-Ying Zhang, Kristina Edfeldt, Manon Oude Nijhuis, Helena Idborg, Magnus Bäck, Joy Roy, Ulf Hedin, Per-Johan Jakobsson, Jon D Laman, Dominique P de Kleijn, Gerard Pasterkamp, Göran K Hansson, Zhong-Qun Yan
OBJECTIVE: The activity of eicosanoid pathways is critical to the inflammatory and immune responses that are associated with the progression of atherosclerosis. Yet, the signals that regulate these pathways are poorly understood. Here, we address whether the innate immune signals of nucleotide-binding oligomerization domain-containing protein (NOD) 2 affect eicosanoids metabolism in atherosclerosis. APPROACH AND RESULTS: Analysis of human carotid plaques revealed that NOD2 was abundantly expressed at both mRNA and protein levels by endothelial cells and macrophages...
September 2013: Arteriosclerosis, Thrombosis, and Vascular Biology
Jae Sam Lee, Joel D Morrisett, Ching-Hsuan Tung
OBJECTIVE: The objective of this study is to develop a method for selective detection of the calcific (hydroxyapatite) component in human aortic smooth muscle cells in vitro and in calcified cardiovascular tissues ex vivo. This method uses a novel optical molecular imaging contrast dye, Cy-HABP-19, to target calcified cells and tissues. METHODS: A peptide that mimics the binding affinity of osteocalcin was used to label hydroxyapatite in vitro and ex vivo. Morphological changes in vascular smooth muscle cells were evaluated at an early stage of the mineralization process induced by extrinsic stimuli, osteogenic factors and a magnetic suspension cell culture...
October 2012: Atherosclerosis
Kiyotaka Nakagawa, Akira Shibata, Tatsuya Saito, Phumon Sookwong, Shunji Kato, Tsuyoshi Tsuduki, Kiminori Matsubara, Teruo Miyazawa
BACKGROUND: Phosphatidylcholine hydroperoxide (PCOOH) is a primary oxidation product of PC, and is markedly accumulated in blood plasma and arterial walls in atherosclerotic animals and humans. The role of PCOOH in the induction of angiogenesis is unknown. METHODS: In this study, we investigated whether PCOOH stimulated angiogenic responses (e.g., vascular endothelial growth factor (VEGF)-induced cell proliferation, migration, and tube formation, and angiogenesis-related gene/protein expression) in human umbilical vein endothelial cells (HUVEC) and in an ex vivo rat aorta model...
December 2011: Biochimica et Biophysica Acta
Agnieszka Czyzewska-Buczyńska, Wojciech Witkiewicz
Atherosclerosis is chronic, inflammatory disease. In artery inflammation main role play cells of the immune system, including lymphocytes and macrophages. This circle of the cells modulating atherogenesis enclose also mast cells. Activated mast cell degranulate and release many types of mediators, including cytokines, chemokins, growth factors, vasoactive substances and proteolytic enzymes. This mediators can modulate inflammatory reaction in artery wall directly, by releasing proinflammatory cytokines or indirectly, influencing the activity of the other cells of the immune system, taking a part in the process of atherogenesis...
2011: Przegla̧d Lekarski
Svitlana Demyanets, Viktoria Konya, Stefan P Kastl, Christoph Kaun, Sabine Rauscher, Alexander Niessner, Richard Pentz, Stefan Pfaffenberger, Kathrin Rychli, Christof E Lemberger, Rainer de Martin, Akos Heinemann, Ihor Huk, Marion Gröger, Gerald Maurer, Kurt Huber, Johann Wojta
OBJECTIVE: Interleukin (IL)-33 is the most recently described member of the IL-1 family of cytokines and it is a ligand of the ST2 receptor. While the effects of IL-33 on the immune system have been extensively studied, the properties of this cytokine in the cardiovascular system are much less investigated. Methods/Results- We show here that IL-33 promoted the adhesion of human leukocytes to monolayers of human endothelial cells and robustly increased vascular cell adhesion molecule-1, intercellular adhesion molecule-1, endothelial selectin, and monocyte chemoattractant protein-1 protein production and mRNA expression in human coronary artery and human umbilical vein endothelial cells in vitro as well as in human explanted atherosclerotic plaques ex vivo...
September 2011: Arteriosclerosis, Thrombosis, and Vascular Biology
Juan Antonio Moreno, Almudena Ortega-Gómez, Sandrine Delbosc, Nathalie Beaufort, Emmanuel Sorbets, Liliane Louedec, Marina Esposito-Farèse, Florence Tubach, Antonino Nicoletti, Philippe Gabriel Steg, Jean-Baptiste Michel, Laurent Feldman, Olivier Meilhac
AIMS: CD163 is a macrophage receptor for haemoglobin-haptoglobin (Hb-Hp) complexes, responsible for the clearance of haemoglobin. We hypothesized that production of soluble CD163 (sCD163) may be due to proleolytic shedding of membrane CD163 by neutrophil elastase, reported to be increased in culprit atherosclerotic plaques. We analysed the relationship between CD163 solubilization and elastase in vitro, in macrophage culture, ex vivo in human atherosclerotic plaque samples, and in vivo, in plasma of patients with coronary artery disease...
January 2012: European Heart Journal
Johan Duchene, Cécile Cayla, Sandrine Vessillier, Ramona Scotland, Kazuo Yamashiro, Florence Lecomte, Irfan Syed, Phuong Vo, Alessandra Marrelli, Costantino Pitzalis, Francesco Cipollone, Joost Schanstra, Jean-Loup Bascands, Adrian J Hobbs, Mauro Perretti, Amrita Ahluwalia
OBJECTIVE: The proinflammatory phenotype induced by low laminar shear stress (LSS) is implicated in atherogenesis. The kinin B1 receptor (B1R), known to be induced by inflammatory stimuli, exerts many proinflammatory effects including vasodilatation and leukocyte recruitment. We investigated whether low LSS is a stimulus for endothelial B1R expression and function. METHODS AND RESULTS: Human and mouse atherosclerotic plaques expressed high level of B1R mRNA and protein...
November 2009: Arteriosclerosis, Thrombosis, and Vascular Biology
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