Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 10 de 10
Filter
Add more filters











Publication year range
1.
Nano Lett ; 24(42): 13293-13299, 2024 Oct 23.
Article in English | MEDLINE | ID: mdl-39361530

ABSTRACT

In biological systems, nanoparticles interact with biomolecules, which may undergo protein corona formation that can result in noncontrolled aggregation. Therefore, comprehending the behavior and evolution of nanoparticles in the presence of biological fluids is paramount in nanomedicine. However, traditional lab-based colloid methods characterize diluted suspensions in low-complexity media, which hinders in-depth studies in complex biological environments. Here, we apply X-ray photon correlation spectroscopy (XPCS) to investigate silica nanoparticles (SiO2) in various environments, ranging from low to high complex biological media. Interestingly, SiO2 revealed Brownian motion behavior, irrespective of the complexity of the chosen media. Moreover, the SiO2 surface and media composition were tailored to underline the differences between a corona-free system from protein corona and aggregates formation. Our results highlighted XPCS potential for real-time nanoparticle analysis in biological media, surpassing the limitations of conventional techniques and offering deeper insights into colloidal behavior in complex environments.


Subject(s)
Nanoparticles , Protein Corona , Silicon Dioxide , Silicon Dioxide/chemistry , Nanoparticles/chemistry , Protein Corona/chemistry , Photons , Colloids/chemistry , Surface Properties
2.
Langmuir ; 39(19): 6823-6836, 2023 05 16.
Article in English | MEDLINE | ID: mdl-37129569

ABSTRACT

To date, much effort has been devoted toward the study of protein corona formation onto large gold nanoparticles (GNPs). However, the protein corona concept breaks down for GNPs in the ultrasmall size regime (<3 nm), and, as a result, our understanding of ultrasmall GNP (usGNP)-protein interactions remains incomplete. Herein, we used anionic usGNPs and six different proteins as model systems to systematically investigate usGNP-protein interactions, with particular focus on the time evolution and long-term behavior of complex formation. The different proteins comprised chymotrypsin (Cht), trypsin (Try), thrombin (Thr), serum albumin (HSA), cytochrome c (Cyt c), and factor XII (FXII). We used a range of biochemical and biophysical methods to estimate binding affinities, determine the effects of usGNPs on protein structure and function, assess the reversibility of any protein structural and functional changes, and evaluate usGNP-protein complex stability. Among the main findings, we observed that prolonged (24 h)─but not short-term (10 min)─interactions between proteins and usGNPs permanently altered protein function, including enzyme activities (Try, Thr, and FXIIa), peroxidase-like activity (Cyt c), and ligand-binding properties (HSA). Remarkably, this occurred without any large-scale loss of the native global conformation, implying time-dependent effects of usGNPs on local protein conformation or dynamics. We also found that both short-(10 min) and long-term (24 h) interactions between proteins and usGNPs yielded short-lived complexes, i.e., there was no time-dependent "hardening" of the interactions at the binding interface as usually seen with large GNPs. The present study increases our fundamental understanding of nano-bio interactions in the ultrasmall size regime, which may assist the safe and effective translation of usGNPs into the clinic.


Subject(s)
Metal Nanoparticles , Protein Corona , Gold/chemistry , Metal Nanoparticles/chemistry , Protein Corona/chemistry , Serum Albumin , Protein Conformation
3.
Colloids Surf B Biointerfaces ; 213: 112387, 2022 May.
Article in English | MEDLINE | ID: mdl-35151044

ABSTRACT

The protein adsorption onto poly(acrylic acid)-block-polystyrene (PAA22-b-PS144) polymersomes has been investigated with regard to structural features, thermodynamic aspects and biological consequences. The light scattering measurements revealed the formation of protein coronas enveloping the polymeric capsules regardless of the chemical nature of the biomacromolecules. The experiments were conducted by using lysozyme, immunoglobulin G - IgG and bovine serum albumin - BSA as model proteins due to their differences concerning size and residual surface charge at physiological pH. The protein adsorption was further confirmed by isothermal titration calorimetry, and the experimental data suggest that the phenomenon is mainly governed by hydrogen bonding and van der Waals interactions. The pre-existing protein layer via the pre-incubation in protein environments notably attenuates the cytotoxicity of the nanomaterial compared to the pristine counterparts. This approach can possibly be extended to different types of assemblies when intermolecular interactions are able to induce protein adsorption and the development of protein coronas around nanoparticles. Such fairly simple method may be convenient to engineer safer nanomaterials towards a variety of biomedical applications when the nanotoxicity is an issue. Additionally, the strategy can possibly be used to tailor the surface properties of nanoparticles by adsorbing specific proteins for targeting purposes.


Subject(s)
Nanoparticles , Nanostructures , Protein Corona , Adsorption , Nanoparticles/chemistry , Protein Corona/chemistry , Serum Albumin, Bovine/chemistry
4.
J Mater Chem B ; 9(8): 2073-2083, 2021 03 04.
Article in English | MEDLINE | ID: mdl-33594396

ABSTRACT

The formation of biomolecular coronas around nanoparticles as soon as they come in contact with biological media is nowadays well accepted. The self-developed biological outer surfaces can affect the targeting capability of the colloidal carriers as well as their cytotoxicity and cellular uptake behavior. In this framework, we explored the structural features and biological consequences of protein coronas around block copolymer assemblies consisting of a common pH-responsive core made by poly[2-(diisopropylamino) ethyl methacrylate] (PDPA) and hydrophilic shells of different chemical natures: zwitterionic poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) or highly hydrophilic poly(ethylene oxide) (PEO) and poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA). We demonstrated the presence of ∼50 nm protein coronas around the nanoparticles regardless of the chemical nature of the polymeric shells. The thickness is understood as the sum of the soft and hard layers and it is the actual interface seen by the cells. Although the soft corona composition is difficult to determine because the proteins are loosely bound to the outer surface of the assemblies, the tightly bound proteins (hard corona) could be identified and quantified. The compositional analysis of the hard corona demonstrated that human serum albumin (HSA), immunoglobulin G (IgG) and fibrinogen are the main components of the protein coronas, and serotransferrin is present particularly in the protein corona of the zwitterionic-stabilized assemblies. The protein coronas substantially reduce the cellular uptake of the colloidal particles due to their increased size and the presence of HSA which is known to reduce nanoparticle-cell adhesion. On the other hand, their existence also reduces the levels of cytotoxicity of the polymeric assemblies, highlighting that protein coronas should not be always understood as artifacts that need to be eliminated due to their positive outputs.


Subject(s)
Mechanical Phenomena , Nanoparticles/chemistry , Protein Corona/chemistry , Cell Adhesion , Humans , Hydrogen-Ion Concentration , Polymers/chemistry , Surface Properties
5.
J Mater Chem B ; 9(2): 428-439, 2021 01 14.
Article in English | MEDLINE | ID: mdl-33367419

ABSTRACT

The use of hybrid nanostructures based on magneto-luminescent properties is a promising strategy for nano-bio applications and theranostics platforms. In this work, we carried out the synthesis and functionalization of iron oxide nanocubes (IONCs) to obtain multifunctional hybrid nanostructures towards biomedical applications. The IONCs were functionalized with tetraethylorthosilicate, thenoyltrifluoroacetone-propyl-triethoxysilane and europium(iii)-dibenzoylmethane complexes to obtain the materials termed as IOCNCs@SiO2, IONCs@SiO2TTA, IONCs@SiO2TTA-Eu and IONCs@SiO2-TTA-Eu-DBM, respectively. Then, the biological interactions of these nanostructures with red blood cells - RBCs (hemolysis) and human blood plasma (protein corona formation) were evaluated. The XPS spectrocopy and EDS chemical mapping analysis showed that each domain is homogeneously occupied in the hybrid material, with the magnetic core at the center and the luminescent domain on the surface of the hybrid nanomaterial with a core@shell like structure. Futhermore, after each functionalization step, the nanomaterial surface charge drastically changed, with critical impact on RBC lysis and corona formation. While IONCs@SiO2 and IONCs@SiO2-TTA-Eu-DBM showed hemolytic properties in a dose-dependent manner, the IONCs@SiO2TTA-Eu did not present any hemolytic effect up to 300 µg mL-1. Protein corona results showed a pattern of selective adsorption of proteins with each surface of the synthesized hybrid materials. However, as a general result, a suppression of hemolysis after protein corona formation in all tests was verified. Finally, this study provides a solid background for further applications of these hybrid magneto-luminescent materials containing new surface functionalities in the emerging field of medical nanobiotechnology.


Subject(s)
Europium/chemistry , Ferric Compounds/chemistry , Nanotechnology/methods , Protein Corona/chemistry , Humans
6.
Bioconjug Chem ; 31(11): 2638-2647, 2020 11 18.
Article in English | MEDLINE | ID: mdl-33169610

ABSTRACT

The success of targeted drug delivery systems still requires a detailed understanding about the biological consequences of self-developed biomolecular coronas around them, since this is the surface that interacts with living cells. Herein, we report the behavior of carbohydrate-decorated amphiphilic nanoparticles in a plasma environment with regard to the formation and biological consequences of the protein corona. Naked amphiphilic nanoparticles were produced through the self-assembly of azido-PEO900-docosanoate molecules, and the coupling of N-acetylglucosamine via click chemistry enabled the fabrication of the corresponding bioactive glyco-nanostructures. Light scattering measurements, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, liquid chromatography-mass spectrometry, and the Pierce BCA protein assay all confirmed the presence of protein coronas around the self-assembled nanoparticles, regardless of the presence of the sugar residues, although it reduces the amount of adsorbed proteins. The protein coronas were formed mainly by human serum albumin, complement proteins, apolipoproteins, immunoglobulins, and proteins involved in the coagulation cascade (fibrinogen and prothrombin). While the presence of these protein coronas significantly reduced cellular uptake of the amphiphilic assemblies, they also notably reduced the cytotoxic and hemolytic effects that result from the contact of the nanoparticles with living cells. Accordingly, we highlight that protein coronas should not always be treated as artifacts that have to be avoided because they can also provide beneficial effects.


Subject(s)
Nanoparticles/chemistry , Protein Corona/chemistry , Adsorption , Chromatography, Liquid/methods , Electrophoresis, Polyacrylamide Gel , HeLa Cells , Humans , Mass Spectrometry/methods , Microscopy, Electron, Transmission
7.
Mater Sci Eng C Mater Biol Appl ; 111: 110850, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32279743

ABSTRACT

The use of noble metal nanoparticles in biomedical and biotechnological applications is nowadays well established. Particularly, silver nanoparticles (AgNPs) were proven to be effective for instance as a biocide agent. They also find applications in tumor therapies and sensing applications being encouraging tools for in-vivo imaging. In this framework, whenever they are in contact with living systems, they are rapidly coated by a protein corona thereby influencing a variety of biological events including cellular uptake, blood circulation lifetime, cytotoxicity and, ultimately, the therapeutic effect. Taking these considerations into account, we have explored the behavior of polymer-coated AgNPs in model protein environments focusing on the self-development of protein coronas. The polymers polyethyleneimine (PEI), polyvinylpyrrolidone (PVP) and poly(2-vinyl pyridine)-b-poly(ethylene oxide) (PEO-b-P2VP) were used as stabilizing agents. The chemical nature of the polymer capping remarkably influences the behavior of the hybrid nanomaterials in protein environments. The PEO-b-P2VP and PVP-stabilized AgNPs are essentially inert to the model proteins adsorption. On the other hand, the PEI-stabilized AgNPs interact strongly with bovine serum albumin (BSA). Nevertheless, the same silver colloids were evidenced to be stable in IgG and lysozyme environments. The BSA adsorption into the PEI-stabilized AgNPs is most probably driven by hydrogen bonding and van der Waals interactions as suggested by isothermal titration calorimetry data. The development of protein coronas around the AgNPs may have relevant implications in a variety of biological events. Therefore, further investigations are currently underway to evaluate the influence of its presence on the cytotoxicity, hemolytic effects and biocide properties of the produced hybrid nanomaterials.


Subject(s)
Colloids/chemistry , Polymers/chemistry , Protein Corona/chemistry , Serum Albumin, Bovine/chemistry , Silver/chemistry , Adsorption , Animals , Calorimetry , Cattle , Chickens , Dynamic Light Scattering , Nanoparticles/ultrastructure , Polyethyleneimine/chemistry , Povidone/chemistry , Spectrometry, Fluorescence , Spectrophotometry, Ultraviolet
8.
Mater Sci Eng C Mater Biol Appl ; 100: 363-377, 2019 Jul.
Article in English | MEDLINE | ID: mdl-30948072

ABSTRACT

The interaction of single-layer graphene oxide (SLGO) and multi-layered graphene oxide (MLGO) with a cell culture medium (i.e. DMEM) was studied by evaluating fetal bovine serum (FBS) protein corona formation towards in vitro nanotoxicity assessment and nanobiointeractions. SLGO and MLGO exhibited different colloidal behavior in the culture medium, which was visualized by cryogenic transmission electron microscopy in situ analysis. Exploring proteomics and bioinformatics tools, 394 and 290 proteins were identified on the SLGO and MLGO hard corona compositions, respectively. From this amount, 115 proteins were exclusively detected on the SLGO and merely 11 on MLGO. SLGO enriched FBS proteins involved in metabolic processes and signal transduction, while MLGO enriched proteins involved in cellular development/structure, and lipid transport/metabolic processes. Such a distinct corona profile is due to differences on surface chemistry, aggregation behavior and the surface area of GO materials. Hydrophilic interactions were found to play a greater role in protein adsorption by MLGO than SLGO. Our results point out implications for in vitro studies of graphene oxide materials concerning the effective dose delivered to cells and corona bioactivity. Finally, we demonstrated the importance of integrating conventional and modern techniques thoroughly to understand the GO-FBS complexes towards more precise, reliable and advanced in vitro nanotoxicity assessment.


Subject(s)
Blood Proteins/chemistry , Culture Media/chemistry , Graphite/chemistry , Nanoparticles/toxicity , Protein Corona/chemistry , Toxicity Tests , Animals , Cattle , Proteomics , Water
9.
ACS Appl Mater Interfaces ; 10(49): 41917-41923, 2018 Dec 12.
Article in English | MEDLINE | ID: mdl-30426737

ABSTRACT

Protein coronas form on the surfaces of nanomaterials in biological fluids. This layer of proteins affects the properties of nanomaterials, altering their behavior and masking engineered functionality. The use of nonfouling moieties reduces or prevents corona formation; however, these ligands typically complicate functionalization. We present here a surface modification strategy for silica nanoparticles using specific molar ratios of zwitterionic and amine moieties. Through proper balance of ligands, we were able to generate particles that featured reactive "handles", while retaining nonfouling properties, high hemocompatibility, and low cytotoxicity.


Subject(s)
Materials Testing , Nanoparticles/chemistry , Protein Corona/chemistry , Silicon Dioxide , Animals , Humans , Mice , NIH 3T3 Cells , Silicon Dioxide/metabolism , Silicon Dioxide/pharmacology
10.
Nanoscale ; 10(7): 3235-3244, 2018 Feb 15.
Article in English | MEDLINE | ID: mdl-29383361

ABSTRACT

Synthetic ultrasmall nanoparticles (NPs) can be designed to interact with biologically active proteins in a controlled manner. However, the rational design of NPs requires a clear understanding of their interactions with proteins and the precise molecular mechanisms that lead to association/dissociation in biological media. Although much effort has been devoted to the study of the kinetics mechanism of protein corona formation on large NPs, the nature of NP-protein interactions in the ultrasmall regime is radically different and poorly understood. Using a combination of experimental and computational approaches, we studied the interactions of a model protein, CrataBL, with ultrasmall gold NPs passivated with p-mercaptobenzoic acid (AuMBA) and glutathione (AuGSH). We have identified this system as an ideal in vitro platform to understand the dependence of binding affinity and kinetics on NP surface chemistry. We found that the structural and chemical complexity of the passivating NP layer leads to quite different association kinetics, from slow and reaction-limited (AuGSH) to fast and diffusion-limited (AuMBA). We also found that the otherwise weak and slow AuGSH-protein interactions measured in buffer solution are enhanced in macromolecular crowded solutions. These findings advance our mechanistic understanding of biomimetic NP-protein interactions in the ultrasmall regime and have implications for the design and use of NPs in the crowded conditions common to all biological media.


Subject(s)
Gold , Metal Nanoparticles/chemistry , Protein Corona/chemistry , Kinetics , Protein Binding
SELECTION OF CITATIONS
SEARCH DETAIL