title: Decreased platelet aggregation by shear stress-stimulated endothelial cells in vitro: description of a method and first results in diabetes.
abstract: The interaction between platelets and endothelium in vivo is a complex phenomenon. Our aim was to develop an in vitro system that mimics the in vivo environment and investigate platelet function in a common pathological condition. Human umbilical vein endothelial cells were used and platelets from 28 type 2 diabetes patients were studied under shear stress conditions. Mean coefficient of variation of platelet aggregation was 10% in dynamic conditions in the presence of endothelium. Endothelial cells increased the concentration of inductor needed to achieve 50% platelet aggregation to adenosine diphosphate from 2.6 ± 1.3 in static conditions to 3.7 ± 1.3 µM in dynamic conditions. A similar pattern was observed when collagen was used for platelet activation. Incubation of endothelium with a nitric oxide inhibitor abolished this effect, indicating platelet inhibitory effect of endothelial cells is nitric oxide mediated. Platelet reactivity of healthy controls was less influenced by the presence of endothelial cells and displayed reduced basal platelet reactivity compared with platelets from diabetes patients. We show that platelet aggregation in diabetes as commonly reported in vitro may not fully reflect the in vivo pathophysiological process. Future studies are warranted to investigate other pathological conditions and analyse the effects of antiplatelet agents using this system.
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy.
authors.forename: Maria S
authors.initials: MS
authors.lastname: De Franceschi
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy Department of Nanomedicine, The Methodist Hospital Research Institute, Houston, TX, USA.
authors.forename: Anna L
authors.initials: AL
authors.lastname: Palange
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Mancuso
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Grande
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Muccari
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy.
authors.forename: Faustina B
authors.initials: FB
authors.lastname: Scavelli
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy.
authors.forename: Concetta
authors.initials: C
authors.lastname: Irace
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy patdis@unicz.it.
authors.forename: Agostino
authors.initials: A
authors.lastname: Gnasso
authors.affiliation: Metabolic Diseases Unit, Department of Clinical and Experimental Medicine, 'Mater Domini' Hospital, 'Magna Græcia' University of Catanzaro, Catanzaro, Italy Department of Chemical Engineering, Imperial College London, South Kensington Campus, London, UK.
authors.forename: Claudio
authors.initials: C
authors.lastname: Carallo
mesh_terms: D001791:Blood Platelet Disorders / Q000150:complications / Q000175:diagnosis* / Q000378:metabolism / Q000503:physiopathology; D002478:Cells, Cultured; D003924:Diabetes Mellitus, Type 2 / Q000150:complications*; D015536:Down-Regulation / Q000187:drug effects; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005240:Feasibility Studies; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D052250:Nitric Oxide Synthase Type III / Q000037:antagonists & inhibitors / Q000378:metabolism; D015539:Platelet Activation / Q000187:drug effects; D010973:Platelet Adhesiveness / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D010979:Platelet Function Tests / Q000295:instrumentation; D015203:Reproducibility of Results; D019323:omega-N-Methylarginine / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D019323:omega-N-Methylarginine; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III
keywords: Platelet aggregation; cell culture; diabetes mellitus; endothelium; flow chamber; shear stress