An environmental route of exposure affects the formation of nanoparticle coronas in blood plasma.

UNLABELLED Nanoparticles (NPs) in contact with biological fluids become covered by a tightly bound layer of proteins, the "protein corona", and it is largely accepted that this corona gives a new identity to NPs in biological milieu. We here consider the exposing scenario of NPs through an environmental route exemplified by the use of hydrophobins, highly adhesive proteins that are secreted into the environment in large quantities by fungi. HFBII of Trichoderma reesei has been used as a model protein and we have shown strong binding to polystyrene NPs of different sizes and surface groups. Hydrophobin coated NPs are shown to strongly increase the stability and the dispersion when exposed to human plasma compared to pristine ones particles. It is also shown that the presence of hydrophobin on the NPs results in an attenuated protein corona formation, in a different corona composition, and we also show that hydrophobin remained strongly associated to the NPs in competition with plasma proteins. As a conclusion we therefore suggest that the route of exposure of nanoparticles strongly affects their surface properties and their possible physiological behavior. SIGNIFICANCE This work shows how a self-assembling protein, class II hydrophobin HFBII, with interesting biocompatible coating properties, strongly adsorbs on polystyrene NPs. HFBII is also shown to reduce aggregation of the NPs in human plasma which can increase their bioavailability with potential use in biomedical applications. The results here are also of significance for understanding possible interactions of NPs with living organisms. Hydrophobins are secreted in large quantities into the environment by fungi and this work shows how the biological environment of NPs determines the surface and colloidal properties of the particles by forming a protein corona, and that the history of the particle environment, here simulated with hydrophobin exposure, affects both plasma protein corona formation and dispersion behavior. This work thus simulates how alternative exposure routes affect nanoparticle properties, important in understanding the biological fate of NPs.

[1]  H. Bayır,et al.  Phosphatidylserine Targets Single-Walled Carbon Nanotubes to Professional Phagocytes In Vitro and In Vivo , 2009, PloS one.

[2]  Kenneth A. Dawson,et al.  Effects of the presence or absence of a protein corona on silica nanoparticle uptake and impact on cells. , 2012, ACS nano.

[3]  Stefan Tenzer,et al.  Nanoparticle size is a critical physicochemical determinant of the human blood plasma corona: a comprehensive quantitative proteomic analysis. , 2011, ACS nano.

[4]  M. Rillig A connection between fungal hydrophobins and soil water repellency , 2005 .

[5]  Kenneth A. Dawson,et al.  Protein–Nanoparticle Interactions , 2008, Nano-Enabled Medical Applications.

[6]  Istvan Toth,et al.  Nanoparticle-induced unfolding of fibrinogen promotes Mac-1 receptor activation and inflammation. , 2011, Nature nanotechnology.

[7]  Minnamari Vippola,et al.  Proteomic characterization of engineered nanomaterial-protein interactions in relation to surface reactivity. , 2011, ACS nano.

[8]  M. Ferrari Cancer nanotechnology: opportunities and challenges , 2005, Nature Reviews Cancer.

[9]  Mark E. Davis,et al.  Nanoparticle therapeutics: an emerging treatment modality for cancer , 2008, Nature Reviews Drug Discovery.

[10]  N. Anderson,et al.  The Human Plasma Proteome , 2002, Molecular & Cellular Proteomics.

[11]  Kenneth A. Dawson,et al.  Transferrin Coated Nanoparticles: Study of the Bionano Interface in Human Plasma , 2012, PloS one.

[12]  Merja Penttilä,et al.  Atomic Resolution Structure of the HFBII Hydrophobin, a Self-assembling Amphiphile* , 2004, Journal of Biological Chemistry.

[13]  Andrzej S Pitek,et al.  Reversible versus irreversible binding of transferrin to polystyrene nanoparticles: soft and hard corona. , 2012, ACS nano.

[14]  Katia Perruccio,et al.  Surface hydrophobin prevents immune recognition of airborne fungal spores , 2009, Nature.

[15]  Hak Soo Choi,et al.  Rapid translocation of nanoparticles from the lung airspaces to the body , 2010, Nature Biotechnology.

[16]  G. Cannon,et al.  Adsorption of a Fungal Hydrophobin Onto Surfaces as Mediated by the Associated Polysaccharide Schizophyllan , 1999 .

[17]  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.

[18]  Merja Penttilä,et al.  Surface adhesion of fusion proteins containing the hydrophobins HFBI and HFBII from Trichoderma reesei , 2002, Protein science : a publication of the Protein Society.

[19]  M. Tolomeo,et al.  Synthesis and antiproliferative activity of 3-(2-chloroethyl)-5-methyl-6-phenyl-8-(trifluoromethyl)-5,6-dihydropyrazolo[3,4-f][1,2,3,5]tetrazepin-4-(3H)-one. , 2015, European journal of medicinal chemistry.

[20]  Stefan Tenzer,et al.  Rapid formation of plasma protein corona critically affects nanoparticle pathophysiology. , 2013, Nature nanotechnology.

[21]  Wolfgang J Parak,et al.  A quantitative fluorescence study of protein monolayer formation on colloidal nanoparticles. , 2009, Nature nanotechnology.

[22]  P. Laaksonen,et al.  Self-assembly of class II hydrophobins on polar surfaces. , 2012, Langmuir : the ACS journal of surfaces and colloids.

[23]  Giulio Caracciolo,et al.  Time evolution of nanoparticle-protein corona in human plasma: relevance for targeted drug delivery. , 2013, Langmuir : the ACS journal of surfaces and colloids.

[24]  Juha Rouvinen,et al.  Crystallization and preliminary X-ray characterization of Trichoderma reesei hydrophobin HFBII. , 2004, Acta crystallographica. Section D, Biological crystallography.

[25]  Marco P Monopoli,et al.  Biomolecular coronas provide the biological identity of nanosized materials. , 2012, Nature nanotechnology.

[26]  H. Wösten,et al.  Role of proteins in soil carbon and nitrogen storage: controls on persistence , 2007 .

[27]  Philip M. Kelly,et al.  Transferrin-functionalized nanoparticles lose their targeting capabilities when a biomolecule corona adsorbs on the surface. , 2013, Nature nanotechnology.

[28]  Claus-Michael Lehr,et al.  Atomic force microscopy and analytical ultracentrifugation for probing nanomaterial protein interactions. , 2012, ACS nano.

[29]  H. Santos,et al.  Intravenous delivery of hydrophobin-functionalized porous silicon nanoparticles: stability, plasma protein adsorption and biodistribution. , 2012, Molecular pharmaceutics.

[30]  Warren C W Chan,et al.  Understanding and controlling the interaction of nanomaterials with proteins in a physiological environment. , 2012, Chemical Society reviews.

[31]  Juha Rouvinen,et al.  Two crystal structures of Trichoderma reesei hydrophobin HFBI—The structure of a protein amphiphile with and without detergent interaction , 2006, Protein science : a publication of the Protein Society.

[32]  W. Kreyling,et al.  The influence of pulmonary surfactant on nanoparticulate drug delivery systems. , 2011, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[33]  Jau-Song Yu,et al.  Comprehensive proteomic analysis of mineral nanoparticles derived from human body fluids and analyzed by liquid chromatography-tandem mass spectrometry. , 2011, Analytical biochemistry.

[34]  M. Penttilä,et al.  The hydrophobins HFBI and HFBII from Trichoderma reesei showing efficient interactions with nonionic surfactants in aqueous two-phase systems. , 2001, Biomacromolecules.

[35]  Iseult Lynch,et al.  What the cell "sees" in bionanoscience. , 2010, Journal of the American Chemical Society.

[36]  M. Linder,et al.  Hydrophobins: Proteins that self assemble at interfaces , 2009 .

[37]  Giulio Caracciolo,et al.  Effect of polyethyleneglycol (PEG) chain length on the bio-nano-interactions between PEGylated lipid nanoparticles and biological fluids: from nanostructure to uptake in cancer cells. , 2014, Nanoscale.

[38]  K. Dawson,et al.  The protein corona of dendrimers: PAMAM binds and activates complement proteins in human plasma in a generation dependent manner , 2012 .

[39]  D. L. Cooper,et al.  High-resolution sizing of monolayer-protected gold clusters by differential centrifugal sedimentation. , 2013, ACS nano.

[40]  Albert Duschl,et al.  Time evolution of the nanoparticle protein corona. , 2010, ACS nano.

[41]  T. Xia,et al.  Understanding biophysicochemical interactions at the nano-bio interface. , 2009, Nature materials.

[42]  Molly M. Stevens,et al.  Emerging techniques for submicrometer particle sizing applied to Stöber silica. , 2012, Langmuir : the ACS journal of surfaces and colloids.

[43]  J. Karp,et al.  Nanocarriers as an Emerging Platform for Cancer Therapy , 2022 .

[44]  Giulio Caracciolo,et al.  Size and charge of nanoparticles following incubation with human plasma of healthy and pancreatic cancer patients. , 2014, Colloids and surfaces. B, Biointerfaces.

[45]  Tiina Nakari-Setälä,et al.  Hydrophobins: the protein-amphiphiles of filamentous fungi. , 2005, FEMS microbiology reviews.

[46]  Iseult Lynch,et al.  Physical-chemical aspects of protein corona: relevance to in vitro and in vivo biological impacts of nanoparticles. , 2011, Journal of the American Chemical Society.

[47]  James L. McGrath,et al.  The influence of protein adsorption on nanoparticle association with cultured endothelial cells. , 2009, Biomaterials.

[48]  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.

[49]  Pauline M Rudd,et al.  The "sweet" side of the protein corona: effects of glycosylation on nanoparticle-cell interactions. , 2015, ACS nano.

[50]  Parag Aggarwal,et al.  Interaction of colloidal gold nanoparticles with human blood: effects on particle size and analysis of plasma protein binding profiles. , 2009, Nanomedicine : nanotechnology, biology, and medicine.