title: Red cell-derived microparticles (RMP) as haemostatic agent.
abstract: Among circulating cell-derived microparticles, those derived from red cells (RMP) have been least well investigated. To exploit potential haemostatic benefit of RMP, we developed a method of producing them in quantity, and here report on their haemostatic properties. High-pressure extrusion of washed RBC was employed to generate RMP. RMP were identified and enumerated by flow cytometry. Their size distribution was assessed by Doppler electrophoretic light scattering analysis (DELSA). Interaction with platelets was studied by platelet aggregometry, and shear-dependent adhesion by Diamed IMPACT-R. Thrombin generation and tissue factor (TF) expression was also measured. The effect of RMP on blood samples of patients with bleeding disorders was investigated ex vivo by thromboelastography (TEG). Haemostatic efficacy in vivo was assessed by measuring reduction of blood loss and bleeding time in rats and rabbits. RMP have mean diameter of 0.45 µm and 50% of them exhibit annexin V binding, a proxy for procoagulant phospholipids (PL). No TF could be detected by flow cytometry. At saturating concentrations of MPs, RMP generated thrombin robustly but after longer delay compared to PMP and EMP. RMP enhanced platelet adhesion and aggregation induced by low-dose ADP or AA. In TEG study, RMP corrected or improved haemostatic defects in blood of patients with platelet and coagulation disorders. RMP reduced bleeding time and blood loss in thrombocytopenic rabbits (busulfan-treated) and in Plavix-treated rats. In conclusion, RMP has broad haemostatic activity, enhancing both primary (platelet) and secondary (coagulation) haemostasis, suggesting potential use as haemostatic agent for treatment of bleeding.
authors.affiliation: Wenche Jy, PhD, University of Miami Miller School of Medicine, 1600 NW 10th Ave., RMSB 7109, R36-A, Miami, FL 33136, USA, Tel.: +1 305 243 6617, Fax: +1 305 243 5957, E-mail: wjy@med.miami.edu.
authors.forename: Wenche
authors.initials: W
authors.lastname: Jy
authors.forename: Max E
authors.initials: ME
authors.lastname: Johansen
authors.forename: Carlos
authors.initials: C
authors.lastname: Bidot
authors.forename: Lawrence L
authors.initials: LL
authors.lastname: Horstman
authors.forename: Yeon S
authors.initials: YS
authors.lastname: Ahn
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D001760:Bleeding Time; D001778:Blood Coagulation Disorders / Q000097:blood / Q000628:therapy*; D002066:Busulfan / Q000008:administration & dosage; D002469:Cell Separation; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D000077144:Clopidogrel; D004195:Disease Models, Animal; D004912:Erythrocytes / Q000378:metabolism*; D005434:Flow Cytometry; D006487:Hemostasis / Q000187:drug effects; D006801:Humans; D008297:Male; D010974:Platelet Aggregation / Q000187:drug effects; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D013916:Thrombelastography; D013917:Thrombin / Q000378:metabolism; D013921:Thrombocytopenia / Q000097:blood / Q000139:chemically induced / Q000628:therapy*; D013925:Thromboplastin / Q000378:metabolism; D013988:Ticlopidine / Q000008:administration & dosage / Q000031:analogs & derivatives
chemical_list: D000244:Adenosine Diphosphate; D013925:Thromboplastin; D000077144:Clopidogrel; D013917:Thrombin; D002066:Busulfan; D013988:Ticlopidine
keywords: Animal bleeding models; RMP–platelet interaction; haemostatic agent; red cell microparticles (RMP); thromboelastography