Interaction of Polystyrene Nanoparticles with Supported Lipid Bilayers: Impact of Nanoparticle Size and Protein Corona.

Polystyrene is one of the most widely used plastics. This article reports on the interaction of 50 and 210 nm polystyrene nanoparticles (PSNPs) with human serum albumin (HSA) and transferrin (Tf), as well as their effect on supported lipid bilayers (SLBs), using experimental and theoretical approaches. Dynamic light scattering (DLS) and atomic force microscopy (AFM) measurements show that the increase in diameter for the PSNP-protein bioconjugates depends on nanoparticle size and type of proteins. The circular dichroism (CD) spectroscopy results demonstrate that the proteins preserve their structures when they interact with PSNPs at physiological temperatures. The quartz crystal microbalance (QCM) technique reveals that PSNPs and their bioconjugates show no strong interactions with SLBs. On the contrary, the molecular dynamics simulations (MDS) show that both proteins bind strongly to the lipid bilayer (SLBs) when compared to their binding to a polystyrene surface model. The interaction is strongly dependent on the protein and lipid bilayer composition. Both the PSNPs and their bioconjugates show no toxicity in human umbilical vein endothelial (HUVEC) cells; however, bare 210 nm PSNPs and 50 nm PSNP-Tf bioconjugates show an increase in reactive oxygen species production. This study may be relevant for assessing the impact of plastics on health.

The impact of cell culture media on the interaction of biopolymer-functionalized gold nanoparticles with cells: mechanical and toxicological properties.

Understanding the nanoparticle-cell interactions in physiological media is vital in determining the biological fate of the nanoparticles (NPs). These interactions depend on the physicochemical properties of the NPs and their colloidal behavior in cell culture media (CCM). Furthermore, the impact of the bioconjugates made by nanoparticle with proteins from CCM on the mechanical properties of cells upon interaction is unknown. Here, we analyzed the time dependent stability of gold nanoparticles (AuNPs) functionalized with citrate, dextran-10, dextrin and chitosan polymers in protein poor- and protein rich CCM. Further, we implemented the high-throughput technology real-time deformability cytometry (RT-DC) to investigate the impact of AuNP-bioconjugates on the cell mechanics of HL60 suspension cells. We found that dextrin-AuNPs form stable bioconjugates in both CCM and have a little impact on cell mechanics, ROS production and cell viability. In contrast, positively charged chitosan-AuNPs were observed to form spherical and non-spherical aggregated conjugates in both CCM and to induce increased cytotoxicity. Citrate- and dextran-10-AuNPs formed spherical and non-spherical aggregated conjugates in protein rich- and protein poor CCM and induced at short incubation times cell stiffening. We anticipate based on our results that dextrin-AuNPs can be used for therapeutic purposes as they show lower cytotoxicity and insignificant changes in cell physiology.

Inhibitory Effect of Epigallocatechin Gallate-Silver Nanoparticles and Their Lysozyme Bioconjugates on Biofilm Formation and Cytotoxicity.

Biofilms are multicellular communities of microbial cells that grow on natural and synthetic surfaces. They have become the major cause for hospital-acquired infections because once they form, they are very difficult to eradicate. Nanotechnology offers means to fight biofilm-associated infections. Here, we report on the synthesis of silver nanoparticles (AgNPs) with the antibacterial ligand epigallocatechin gallate (EGCG) and the formation of a lysozyme protein corona on AgNPs, as shown by UV-vis, dynamic light scattering, and circular dichroism analyses. We further tested the activity of EGCG-AgNPs and their lysozyme bioconjugates on the viability of Bacillus subtilis cells and biofilm formation. Our results showed that, although EGCG-AgNPs presented no antibacterial activity on planktonic B. subtilis cells, they inhibited B. subtilis biofilm formation at concentrations larger than 40 nM, and EGCG-AgNP-lysozyme bioconjugates inhibited biofilms at concentrations above 80 nM. Cytotoxicity assays performed with human cells showed a reverse trend, where EGCG-AgNPs barely affected human cell viability while EGCG-AgNP-lysozyme bioconjugates severely hampered viability. Our results therefore demonstrate that EGCG-AgNPs may be used as noncytotoxic antibiofilm agents.

Changing surface properties of artificial lipid membranes at the interface with biopolymer coated gold nanoparticles under normal and redox conditions.

Gold nanoparticles (NPs) functionalized with biopolymers are increasingly effective in drug-delivery applications. Here, we investigated how chitosan coated NPs and dextran-10 coated NPs regulate their action on DMPC bilayer under normal and stress conditions. We found that chitosan-coated NPs interact with lipid membrane in an intermittent manner, causing lipid loss and partial membrane rupture, while dextran-10 coated NPs mostly induced complete rupture as observed by quartz crystal microbalance. In-situ atomic force microscopy imaging showed that chitosan-treated membranes have a higher surface roughness than those treated with dextran-10. Treatment with 1 µM nitric oxide (NO) radical caused the release of chitosan ligand from the surface of gold NPs (reduced stability) and its aggregation, but the functionality seemed less influenced. Dextran-10 ligand showed no such behavior, while its action was only delayed. Our findings give insights into possible challenges faced by NPs in-situ and show environment dependent effects of NPs on membranes.

Biopolymer-coated gold nanoparticles inhibit human insulin amyloid fibrillation.

Deposits of protein misfolding and/or aggregates are a pathological hallmark of amyloid-related diseases. For instance, insulin amyloid fibril deposits have been observed in patients with insulin-dependent diabetes mellitus after insulin administration. Here, we report on the use of AuNPs functionalized with linear- (i.e. dextrin and chitosan) and branched- (i.e. dextran-40 and dextran-10) biopolymers as potential agents to inhibit insulin fibril formation. Our dynamic light scattering analyses showed a size decrease of the amyloid fibrils in the presence of functionalized AuNPs. Circular dichroism spectroscopy as well as enzyme-linked immunosorbent assay data demonstrated that the secondary structural transition from α-helix to ß-sheet (which is characteristic for insulin amyloid fibril formation) was significantly suppressed by all biopolymer-coated AuNPs, and in particular, by those functionalized with linear biopolymers. Both transmission electron microscopy and atomic force microscopy analyses showed that the long thick amyloid fibrils formed by insulin alone become shorter, thinner or cluster when incubated with biopolymer-coated AuNPs. Dextrin- and chitosan-coated AuNPs were found to be the best inhibitors of the fibril formation. Based on these results, we propose a mechanism for the inhibition of insulin amyloid fibrils: biopolymer-coated AuNPsstrongly interact with the insulin monomers and inhibit the oligomer formation as well as elongation of the protofibrils.Moreover, cytotoxicity experiments showed that AuNP-insulin amyloid fibrils are less toxic compared to insulin amyloid fibrils alone. Our results suggest that both dextrin- and chitosan-AuNPs could be used as therapeutic agents for the treatment of amyloid-related disorders.

H-Bonding-mediated binding and charge reorganization of proteins on gold nanoparticles.

Once introduced into the human body, nanoparticles often interact with blood proteins, which in turn undergo structural changes upon adsorption. Although protein corona formation is a widely studied phenomenon, the structure of proteins adsorbed on nanoparticles is far less understood. We propose a model to describe the interaction between human serum albumin (HSA) and nanoparticles (NPs) with arbitrary coatings. Our model takes into account the competition between protonated and unprotonated polymer ends and the curvature of the NPs. To this end, we explored the effects of surface ligands (citrate, PEG-OMe, PEG-NH2, PEG-COOH, and glycan) on gold nanoparticles (AuNPs) and the pH of the medium on structural changes in the most abundant protein in blood plasma (HSA), as well as the impact of such changes on cytotoxicity and cellular uptake. We observed a counterintuitive effect on the ζ-potential upon binding of negatively charged HSA, while circular dichroism spectroscopy at various pH values showed an unexpected pattern in the reduction of α-helix content, as a function of surface chemistry and curvature. Our model qualitatively reproduces the decrease in α-helix content, thereby offering a rationale based on particle curvature. The simulations quantitatively reproduce the charge inversion measured experimentally through the ζ-potential of the AuNPs in the presence of HSA. Finally, we found that AuNPs with adsorbed HSA display lower toxicity and slower cell uptake rates, compared to functionalized systems in the absence of protein. Our study allows examining and explaining the conformational dynamics of blood proteins triggered by NPs and corona formation, thereby opening new avenues toward designing safer NPs for drug delivery and nanomedical applications.

Methylene Blue-Loaded Upconverting Hydrogel Nanocomposite: Potential Material for Near-Infrared Light-Triggered Photodynamic Therapy Application.

The property of upconverting nanoparticles to convert the low-energy near-infrared (NIR) light into high-energy visible light has made them a potential candidate for various biomedical applications including photodynamic therapy (PDT). In this work, we show how a surface functionalization approach on the nanoparticle can be used to develop a nanocomposite hydrogel which can be of potential use for the PDT application. The upconverting hydrogel nanocomposite was synthesized by reacting 10-undecenoic acid-capped Yb3+/Er3+-doped NaYF4 nanoparticles with the thermosensitive N-isopropylacrylamide monomer. The formation of hydrogel was completed within 15 min and hydrogel nanocomposites showed strong enhancement in the visible light emission compared to the emission obtained from 10-undecenoic acid-capped Yb3+/Er3+-doped NaYF4 nanoparticles via the upconversion process (under 980 nm laser excitation). The upconverting hydrogel nanocomposites displayed high swelling behavior in water because of their porous nature. The porous structure ensured a higher loading of methylene blue dye (∼78% in 1 h) into the upconverting hydrogel, which was achieved via the swelling diffusion phenomenon. Upon excitation with the NIR light, the visible light emitted from the hydrogel activated the photosensitizer methylene blue which generated reactive oxygen species. Our results were able to show that the methylene blue-loaded composite hydrogel can be a potential platform for the future of NIR-triggered PDT in skin cancer treatment.

Synthesis of Upconverting Hydrogel Nanocomposites Using Thiol-Ene Click Chemistry: Template for the Formation of Dendrimer-Like Gold Nanoparticle Assemblies.

The synthesis of upconverting hydrogel nanocomposites by base-catalyzed thiol-ene click reaction between 10-undecenoic acid capped Yb(3+)/Er(3+)-doped NaYF4 nanoparticles and pentaerythritol tetrakis(3-mercaptopropionate) (PETMP) as tetrathiol monomer is reported. This synthetic strategy for nanocomposite gels is quite different from works where usually the preformed gels are mixed with the nanoparticles. Developing nanocomposites by surface modification of capping ligands would allow tuning and controlling of the separation of the nanoparticles inside the gel network. The hydrogel nanocomposites prepared by thiol-ene click reaction show strong enhancement in luminescence intensity compared to 10-undecenoic acid-capped Yb(3+)/Er(3+)-doped NaYF4 nanoparticles through the upconversion process (under 980 nm laser excitation). The hydrogel nanocomposites display strong swelling characteristics in water resulting in porous structures. Interestingly, the resulting nanocomposite gels act as templates for the synthesis of dendrimer-like Au nanostructures when HAuCl4 is reduced in the presence of the nanocomposite gels.

Methyl oleate-capped upconverting nanocrystals: a simple and general ligand exchange strategy to render nanocrystals dispersible in aqueous and organic medium.

We report a simple and general ligand exchange strategy to functionalize the nanocrystals with both hydrophobic and hydrophilic ligands. This is achieved by first capping the Er/Yb-doped NaYF4 nanocrystals with a weak ligand such as methyl oleate and subsequently ligand exchanged with various organic ligands which can strongly coordinate to the surface of the nanocrystals. The method involves only a simple stirring or sonication of the nanocrystals dispersion with the ligands of interest. Dicarboxylic acids such as sebacic acid, adipic acid, succinic acid, and malonic acid-functionalized nanocrystals which are difficult to achieve via thermal decomposition method were easily prepared by this ligand exchange strategy. In addition, low boiling point ligands like hexanoic acid can easily be coated over the surface of the Er/Yb-doped NaYF4 nanocrystals. Both size and shape of the nanocrystals were preserved after the ligand exchange process. The methyl oleate-capped Er/Yb-doped NaYF4 nanocrystals display strong upconversion emission after ligand exchanged with hydrophobic and hydrophilic molecules. The high stability of the nanocrystals after ligand exchange process is verified by performing time-dependent luminescent measurements at different pH, buffers, etc.

Eu3+ ions as an optical probe to follow the growth of colloidal ZnO nanostructures.

The colloidal growth of ZnO exhibits interesting dynamics, which is generally probed using absorbance measurements. Here, we have shown that the sharp luminescent signals from the Eu(3+) ions act as a potential luminescent spectral probe to follow the growth of ZnO nanostructures. The Eu(3+)-doped ZnO nanocrystals were synthesized by a colloidal method. The asymmetry ratio calculated from the Eu(3+) emission intensity peaks ((5)D0 → (7)F2/(5)D0 → (7)F1) gradually decrease with the increase in the size of the ZnO nanostructures. This is attributed to the increase in the surface related defects as the size of the ZnO nanocrystals is increased. The above result is supported by controlling the growth of the ZnO nanocrystals with capping ligands. The Eu(3+) luminescence intensity hardly is affected upon ligand capping. Additional experiments such as lifetime measurements and photocatalytic activity of ZnO strongly indicate that Eu(3+) can be used as a potential tool to follow the growth of colloidal ZnO nanostructures. We believe the study can be extended to understand the growth mechanism of several other colloidal nanostructures.

Ricinoleic Acid-Capped Upconverting Nanocrystals: An Ideal Capping Ligand to Render Nanocrystals Water Dispersible.

Cap in hand: Ricinoleic acid (RA) replaces oleic acid as capping ligand in the synthesis of upconverting nanocrystals in the size range of 10 nm. The presence of hydroxyl groups near the double bond in RA is advantageous in hydroxylation of the bond, and the size of the nanocrystals is preserved. The small size, high water dispersibility, and strong upconversion from the nanocrystals could be utilized in sensing and bioimaging applications.