Abstract
Nanoparticles experience increasing interest for a variety of medical and pharmaceutical applications. When exposing nanomaterials, e.g., magnetic iron oxide nanoparticles (MNP), to human blood, a protein corona consisting of various components is formed immediately. The composition of the corona as well as its amount bound to the particle surface is dependent on different factors, e.g., particle size and surface charge. The actual composition of the formed protein corona might be of major importance for cellular uptake of magnetic nanoparticles. The aim of the present study was to analyze the formation of the protein corona during in vitro serum incubation in dependency of incubation time and temperature. For this, MNP with different shells were incubated in fetal calf serum (FCS, serving as protein source) within a water bath for a defined time and at a defined temperature. Before and after incubation the particles were characterized by a variety of methods. It was found that immediately (seconds) after contact of MNP and FCS, a protein corona is formed on the surface of MNP. This formation led to an increase of particle size and a slight agglomeration of the particles, which was relatively constant during the first minutes of incubation. A longer incubation (from hours to days) resulted in a stronger agglomeration of the FCS incubated MNP. Quantitative analysis (gel electrophoresis) of serum-incubated particles revealed a relatively constant amount of bound proteins during the first minutes of serum incubation. After a longer incubation (>20 min), a considerably higher amount of surface proteins was determined for incubation temperatures below 40 °C. For incubation temperatures above 50 °C, the influence of time was less significant which might be attributed to denaturation of proteins during incubation. Overall, analysis of the molecular weight distribution of proteins found in the corona revealed a clear influence of incubation time and temperature on corona composition.
Highlights
Magnetic nanoparticles (MNP) represent perfectly suitable materials for a variety of biomedical and biotechnological applications
Our MRX investigations show that immediately after contact of MNP with a protein source (FCS), a protein corona is formed on the particle surface
Independent of the used polymer shells used for MNP (DEAE, CMD, dextran, poly(tert-butoxycarbonyl acrylic acid) (PtBAA)), zeta potentials from −30 to −40 mV were found after serum incubation
Summary
Magnetic nanoparticles (MNP) represent perfectly suitable materials for a variety of biomedical and biotechnological applications. Upon application of nanoparticles into biological media (e.g., whole-blood or plasma), the formation of a protein “corona” around the particles takes place immediately. Lundquist and coworkers [5] found that for a fixed type of material, the biologically active proteins in the corona are strongly determined by the size as well as the zeta potential of the particles. It is well-known that the adsorption of blood serum proteins to particles is time-dependent [6]. Comprehensive review articles about the influence of different nanoparticle parameters (e.g., composition, size, shape, crystallinity, surface area, surface defects, charge, roughness, transfer capability, and hydrophobicity/hydrophilicity) on the corona composition were published in the past years by several authors [4, 8,9,10,11,12,13]
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