RESUMEN
BACKGROUND: Investigations were performed to determine whether poly-N-acetyl glucosamine (p-GlcNAc) induces hemostasis by the activation of platelets. METHODS: Platelets were isolated from human blood, fixed in the presence poly-N-acetyl glucosamine fibers, and visualized with scanning electron microscopy. Platelet activation surface markers were measured by fluorescence multiphoton microscopy. Platelet aggregation in the presence of p-GlcNAc fibers and integrin receptor blockers was measured. RESULTS: Scanning electron microscopy indicated that contact of platelets with poly-N-acetyl glucosamine fibers resulted in platelet activation. Fluorescent microscopy showed that contact of platelets with the marine polymer increased intracellular levels of free calcium and resulted in surface exposure of platelet phosphatidylserine, P selectin, and the alphaIIbbeta3 integrin. Antibody inhibitors of the platelet alphaIIbbeta3 integrin inhibited p-GlcNAc to stimulate fibrin polymerization. CONCLUSION: Poly-N-acetyl glucosamine fiber material promotes hemostasis by the activation of platelets.
Asunto(s)
Acetilglucosamina/farmacología , Plaquetas/efectos de los fármacos , Quitina/análogos & derivados , Hemostáticos/farmacología , Activación Plaquetaria/efectos de los fármacos , Acetilglucosamina/química , Plaquetas/ultraestructura , Calcio/fisiología , Quitina/química , Quitina/farmacología , Quitosano , Evaluación Preclínica de Medicamentos , Fibrina/efectos de los fármacos , Hemostasis Quirúrgica/métodos , Hemostáticos/química , Humanos , Integrina alfa2 , Integrina alfa5/efectos de los fármacos , Integrina alfa6/efectos de los fármacos , Integrina beta3/efectos de los fármacos , Líquido Intracelular/efectos de los fármacos , Glicoproteínas de Membrana/efectos de los fármacos , Microscopía Electrónica de Rastreo , Microscopía de Fluorescencia por Excitación Multifotónica , Selectina-P/efectos de los fármacos , Fosfatidilserinas/fisiología , Activación Plaquetaria/fisiología , Adhesividad Plaquetaria/efectos de los fármacos , Complejo GPIb-IX de Glicoproteína Plaquetaria/efectos de los fármacos , Glicoproteína IIb de Membrana Plaquetaria/efectos de los fármacos , Espectrofotometría , Factores de TiempoRESUMEN
Polysaccharides are becoming increasingly developed as therapeutics and medical products, as the new field of Glycomics expands. Glycosaminoglycans that contain N-acetyl glucosamine constituents have been the focus of research leading to medical devices. A new hemostatic bandage, the Syvek Patch, has been introduced in the recent past for the control of bleeding at vascular access sites in interventional cardiology and radiology procedures. This product consists of poly-N-acetyl glucosamine (pGlcNAc) isolated in a unique fiber crystalline structural form from the large-scale culture and processing of a marine diatom. The Syvek pGlcNAc fiber material has chemical, physical, and biological properties that result in its favorable performance as a hemostat. Two new products, the Clo-Sur PAD and ChitoSeal, have recently become available also as patch hemostats. These two products both use chitosan, another N-acetyl glucosamine containing glycosaminoglycan, as their active ingredient. Structural, chemical, and biological comparisons of Syvek pGlcNAc and chitosan reveal a number of important differences. Syvek pGlcNAc fibers contain approximately 50 fully acetylated, high molecular weight pGlcNAc molecules in a crystalline, three-dimensional beta structure array, and are insoluble. Chitosan is a low molecular weight mixed amorphous cationic polymer with no regular structure as a solid, and is water-soluble taking on a random coil configuration when in solution. These structural dissimilarities result in differences in the hemostatic properties of the two materials. Syvek pGlcNAc is able to significantly reduce the in vitro fibrin clot formation time of platelet-rich plasma samples and has the ability to cause aggregation of red blood cells in vitro. Chitosan is no better than gauze or other controls in these in vitro assays. The Syvek Patch is able to control the bleeding and cause hemostasis in a coagulopathic swine spleen-bleeding animal model 100% of the time, whereas Clo-Sur PAD was completely unsuccessful (0%) and ChitoSeal (25%) was worse than a gauze pad control (50%) in the same model. Syvek pGlcNAc fibers have structural and chemical properties that provide a unique basis for their ability to interact with blood components to cause hemostasis. Chitosan does not have the same properties and capabilities.
Asunto(s)
Acetilglucosamina/farmacología , Quitina/análogos & derivados , Quitina/química , Hemostasis/efectos de los fármacos , Hemostáticos/farmacología , Acetilglucosamina/química , Animales , Anticoagulantes/química , Anticoagulantes/farmacología , Quitosano , Evaluación Preclínica de Medicamentos , Hemostasis/fisiología , Hemostáticos/química , Humanos , PorcinosRESUMEN
Recombinant human interleukin-11 (rhIL-11), a glycoprotein 130 (gp130)-signaling cytokine approved for treatment of thrombocytopenia, also raises von Willebrand factor (VWF) and factor VIII (FVIII) by an unknown mechanism. Desmopressin (1-deamino-8-d-arginine vasopressin [DDAVP]) releases stored VWF and FVIII and is used for treatment of VWF and FVIII deficiencies. To compare the effect of these 2 agents, heterozygous von Willebrand disease (VWD) and normal dogs were treated with either rhIL-11 (50 microg/kg/d subcutaneously x 7 days) or DDAVP (5 microg/kg/d intravenously x 7 days). The rhIL-11 produced a gradual and sustained elevation of VWF and FVIII levels in both heterozygous VWD and normal dogs while DDAVP produced a rapid and unsustained increase. Importantly, rhIL-11 treatment produced a 2.5- to 11-fold increase in VWF mRNA in normal canine heart, aorta, and spleen but not in homozygous VWD dogs, thus identifying a mechanism for elevation of plasma VWF in vivo. Moreover, dogs pretreated with rhIL-11 retain a DDAVP-releasable pool of VWF and FVIII, suggesting that rhIL-11 does not significantly alter trafficking of these proteins to or from storage pools. The half-life of infused VWF is unchanged by rhIL-11 in homozygous VWD dogs. These results show that rhIL-11 and DDAVP raise plasma VWF by different mechanisms. Treatment with rhIL-11 with or without DDAVP may provide an alternative to plasma-derived products for some VWD and hemophilia A patients if it is shown safe in clinical trials.