A hyperspectral and toxicological analysis of protein corona impact on silver nanoparticle properties, intracellular modifications, and macrophage activation

Abstract

The inevitable adsorption of biomolecules on nanomaterials results in the formation of a protein corona (PC), which modifies the nanoparticle (NP)–cell interface resulting in modified uptake, activity, clearance, and toxicity. While the physicochemical properties of the NP govern the composition of PC, the formation of PC in turn alters the characteristics of the NP by imparting a new unique “biological” identity. To assess how the PC influences AgNP properties, intracellular modifications, and cellular responses, we utilized a combination of hyperspectral and toxicological analyses. AgNPs were coated with a complex PC (multiple proteins, eg, 10% fetal bovine serum) or a simple PC (single protein, eg, bovine serum albumin [BSA]) and evaluated by hyperspectral and dynamic light scattering for modifications in AgNP properties. Mouse macrophages were exposed to AgNPs with PCs and examined for differences in uptake, cytotoxicity, and cell activation. Hyperspectral imaging revealed intracellular modifications to AgNPs that were found to spectrally match alterations in AgNPs following incubation in lysosomal fluid. Addition of the PC influenced AgNP uptake and cytotoxicity; however, hydrodynamic size and surface charge did not contribute to these responses. Assessments of all endpoints demonstrated differences between complex and BSA PC, suggesting that these responses are not purely driven by the primary protein component of the complex PC (ie, BSA). Alterations in cellular–NP uptake/interactions may be driven through cell surface receptor recognition of protein constituents that make up the PC rather than the physicochemical differences in AgNPs.

Tissue distribution and acute toxicity of silver after single intravenous administration in mice: nano-specific and size-dependent effects

“…The low dissolution of AgNPs when incubated in mouse serum is in accordance with other studies, which put forward that following introduction into a physiological environment the ability of AgNPs to release Ag + is likely inhibited due to the formation of the protein corona [ 39 ]. Interaction of protein thiol groups with the charged surface of AgNPs in a medium with a high ionic strength is at the basis of the sulfidation process that leads to an extensive decrease of the dissolution rate in vitro , causing also the formation of nanobridges between particles, and has the potential to stabilize them in vivo [ 40 ].…”

Mass spectrometry-based proteomics for system-level characterization of biological responses to engineered nanomaterials

“…Consequently, the physicochemical properties of ENMs are significantly affected by the associated proteins [12]. The identities and nature of proteins forming the corona have been under intense investigation, especially through mass spectrometry (MS)-based approaches [13–18]. Moreover, the ability of various ENMs in initiating cellular oxidative stress or generating reactive oxygen species (ROS) has been suggested as a major paradigm of adaptive and toxic cellular responses to ENMs [19–22] (Figure 1a).…”

Variations in biocorona formation related to defects in the structure of single walled carbon nanotubes and the hyperlipidemic disease state

“…For example, serum albumin is the primary component of the BC that forms on silver nanoparticles due to its abundance 30 . However, when the inflammatory response of macrophages was evaluated following exposure to either silver nanoparticles with a BC that only consisted of albumin or a complex BC, differences were evident 34 . This suggests that even minor changes in the identity of the BC can elicit significant changes in biological responses.…”