Nanoparticle size and surface charge determine effects of PAMAM dendrimers on human platelets in vitro.

Blood platelets are essential in maintaining hemostasis. Various materials can activate platelets and cause them to aggregate. Platelet aggregation in vitro is often used as a marker for materials' thrombogenic properties, and studying nanomaterial interaction with platelets is an important step toward understanding their hematocompatibility. Here we report evaluation of 12 formulations of PAMAM dendrimers varying in size and surface charge. Using a cell counter based method, light transmission aggregometry and scanning electron microscopy, we show that only large cationic dendrimers, but not anionic, neutral or small cationic dendrimers, induce aggregation of human platelets in plasma in vitro. The aggregation caused by large cationic dendrimers was proportional to the number of surface amines. The observed aggregation was not associated with membrane microparticle release, and was insensitive to a variety of chemical and biological inhibitors known to interfere with various pathways of platelet activation. Taken in context with previously reported studies, our data suggest that large cationic PAMAM dendrimers induce platelet aggregation through disruption of membrane integrity.

[1]  M. Gelderman,et al.  Carbon nanotubes activate blood platelets by inducing extracellular Ca2+ influx sensitive to calcium entry inhibitors. , 2009, Nano letters.

[2]  Thommey P. Thomas,et al.  Synthesis and in vitro testing of J591 antibody-dendrimer conjugates for targeted prostate cancer therapy. , 2004, Bioconjugate chemistry.

[3]  E. W. Meijer,et al.  Dendrimers: relationship between structure and biocompatibility in vitro, and preliminary studies on the biodistribution of 125I-labelled polyamidoamine dendrimers in vivo. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[4]  Sara Linse,et al.  Understanding the nanoparticle–protein corona using methods to quantify exchange rates and affinities of proteins for nanoparticles , 2007, Proceedings of the National Academy of Sciences.

[5]  Gustavo A. Abraham,et al.  Antithrombogenic properties of bioconjugate streptokinase-polyglycerol dendrimers , 2006, Journal of materials science. Materials in medicine.

[6]  D. Benaki,et al.  Bare and protein-conjugated Fe3O4 ferromagnetic nanoparticles for utilization in magnetically assisted hemodialysis: biocompatibility with human blood cells , 2008, Nanotechnology.

[7]  Lang Tran,et al.  Surface derivatization state of polystyrene latex nanoparticles determines both their potency and their mechanism of causing human platelet aggregation in vitro. , 2011, Toxicological sciences : an official journal of the Society of Toxicology.

[8]  J. Lieske,et al.  Biologic nanoparticles and platelet reactivity. , 2009, Nanomedicine.

[9]  Seungpyo Hong,et al.  Interaction of polycationic polymers with supported lipid bilayers and cells: nanoscale hole formation and enhanced membrane permeability. , 2006, Bioconjugate chemistry.

[10]  Jie Xue,et al.  Biomimetic glycoliposomes as nanocarriers for targeting P-selectin on activated platelets. , 2007, Bioconjugate chemistry.

[11]  Anil K Patri,et al.  Dendrimer-induced leukocyte procoagulant activity depends on particle size and surface charge. , 2012, Nanomedicine.

[12]  A. Baird,et al.  Circulating endothelial microparticles in acute ischemic stroke: a link to severity, lesion volume and outcome , 2006, Journal of thrombosis and haemostasis : JTH.

[13]  A. Lázníčková,et al.  Dendrimers: Analytical characterization and applications. , 2009, Bioorganic chemistry.

[14]  Hidenori Suzuki,et al.  New strategy of platelet substitutes for enhancing platelet aggregation at high shear rates: cooperative effects of a mixed system of fibrinogen γ-chain dodecapeptide- or glycoprotein Ibα-conjugated latex beads under flow conditions , 2006, Journal of Artificial Organs.

[15]  I. Andricioaei,et al.  Poly(amidoamine) dendrimers on lipid bilayers II: Effects of bilayer phase and dendrimer termination. , 2008, The journal of physical chemistry. B.

[16]  Martin Holzer,et al.  Single-walled carbon nanotubes activate platelets and accelerate thrombus formation in the microcirculation. , 2010, Toxicology.

[17]  Donald A. Tomalia,et al.  In quest of a systematic framework for unifying and defining nanoscience , 2009, Journal of Nanoparticle Research.

[18]  M. Morandi,et al.  Nanoparticle‐induced platelet aggregation and vascular thrombosis , 2005, British journal of pharmacology.

[19]  M. Gelderman,et al.  Flow cytometric analysis of cell membrane microparticles. , 2008, Methods in molecular biology.

[20]  Y. Ohtsuka,et al.  Mechanisms of blood coagulation induced by latex particles and the roles of blood cells. , 1990, Biomaterials.

[21]  Jian Qin,et al.  The importance of an endotoxin-free environment during the production of nanoparticles used in medical applications. , 2006, Nano letters.

[22]  G. Zbinden,et al.  Assessment of thrombogenic potential of liposomes. , 1989, Toxicology.

[23]  Eleonore Fröhlich,et al.  The role of nanoparticle size in hemocompatibility. , 2009, Toxicology.

[24]  K. Jacobson,et al.  Application of the functionalized congener approach to dendrimer-based signaling agents acting through A2A adenosine receptors , 2008, Purinergic Signalling.

[25]  Hamidreza Ghandehari,et al.  Size and surface charge significantly influence the toxicity of silica and dendritic nanoparticles , 2012, Nanotoxicology.

[26]  Y. Ohtsuka,et al.  Platelet aggregation induced by latex particles. I. Effects of size, surface potential and hydrophobicity of particles. , 1989, Biomaterials.

[27]  Clinton F Jones,et al.  In vitro assessments of nanomaterial toxicity. , 2009, Advanced drug delivery reviews.

[28]  Hisataka Kobayashi,et al.  Macromolecular MRI contrast agents with small dendrimers: pharmacokinetic differences between sizes and cores. , 2003, Bioconjugate chemistry.

[29]  J. B. Hall,et al.  Characterization of nanoparticles for therapeutics. , 2007, Nanomedicine.

[30]  Seungpyo Hong,et al.  Interaction of poly(amidoamine) dendrimers with supported lipid bilayers and cells: hole formation and the relation to transport. , 2004, Bioconjugate chemistry.

[31]  J. Šimák.  Nanotoxicity in Blood: Effects of Engineered Nanomaterials on Platelets , 2009 .

[32]  Marina A Dobrovolskaia,et al.  Evaluation of nanoparticle immunotoxicity. , 2009, Nature nanotechnology.

[33]  S. Howorka,et al.  Nanoscale protein pores modified with PAMAM dendrimers. , 2007, Journal of the American Chemical Society.

[34]  Vivien Marx,et al.  Poised to branch out , 2008, Nature Biotechnology.

[35]  A. Mark,et al.  Coarse grained model for semiquantitative lipid simulations , 2004 .

[36]  Kenneth A. Dawson,et al.  Nanoparticle size and surface properties determine the protein corona with possible implications for biological impacts , 2008, Proceedings of the National Academy of Sciences.

[37]  J. Oh,et al.  Blood Compatibility of Cetyl Alcohol/Polysorbate-Based Nanoparticles , 2005, Pharmaceutical Research.

[38]  J. Chung,et al.  Silver nanoparticles enhance thrombus formation through increased platelet aggregation and procoagulant activity , 2011, Nanotoxicology.

[39]  Donald A. Tomalia,et al.  The dendritic state , 2005 .

[40]  Sara Linse,et al.  The nanoparticle-protein complex as a biological entity; a complex fluids and surface science challenge for the 21st century. , 2007, Advances in colloid and interface science.

[41]  R. Larson,et al.  Molecular dynamics simulations of PAMAM dendrimer-induced pore formation in DPPC bilayers with a coarse-grained model. , 2006, The journal of physical chemistry. B.