Nanoparticles In Biological Media Research Articles

Probing Protein Corona Formation around Gold Nanoparticles: Effects of Surface Coating.

There has been much interest in integrating various inorganic nanoparticles (nanoscale colloids) in biology and medicine. However, buildup of a protein corona around the nanoparticles in biological media, driven by nonspecific interactions, remains a major hurdle for the translation of nanomedicine into clinical applications. In this study, we investigate the interactions between gold nanoparticles and serum proteins using a series of dihydrolipoic acid (DHLA)-based ligands. We employed gel electrophoresis combined with UV-vis absorption and dynamic light scattering to correlate protein adsorption with the nature and size of the ligand used. For instance, we found that AuNPs capped with DHLA alone promote nonspecific protein adsorption. In comparison, capping AuNPs with polyethylene glycol- or zwitterion-appended DHLA essentially prevents corona formation, regardless of ligand charge and size. Our results highlight the crucial role of surface chemistry and core material in protein corona formation and offer valuable information for the design of colloidal nanomaterials for biological applications.

Synthesis of cRGD peptide cluster-decorated NIR-fluorescent PISA-RAFT nanoparticles targeting integrin expressing cells

Biocompatible PISA-RAFT nanoparticles were synthesized with NIR-fluorescent monomers in the core and cRGD peptide clusters at the outer surface. These original nanoprobes selectively labeled integrin-presenting cells.

Fixation and Visualization of Full Protein Corona on Lipid Surface of Composite Nanoconstruction.

Spontaneous sorption of proteins on the nanoparticles' surface leads to the fact that nanoparticles in biological media are always enveloped by a layer of proteins-the protein corona. Corona proteins affect the properties of nanoparticles and their behavior in a biological environment. In this regard, knowledge about the composition of the corona is a necessary element for the development of nanomedicine. Because proteins have different sorption efficacy, isolating particles with a full corona and characterizing the full corona is challenging. In this study, we propose a photo-activated cross-linker for full protein corona fixation. We believe that the application of our proposed approach will make it possible to capture and visualize the full corona on nanoparticles coated with a lipid shell.

Methods of Atomic Spectroscopy in Studying Properties and the Behavior of Nanoscale Magnetic Materials in Biological Systems

In this review, we provide an overview of methods for synthesizing magnetic nanoparticles (MNPs) with potential applications to biomedical research. We explore how the structure and properties of these particles are related to their diverse uses in medical diagnostics and bioanalysis. Special emphasis is placed on MNPs containing noble metals, which serve as biomarkers or active agents. Specifically, we focus on the application of direct and combined methods of atomic spectroscopy (ETAAS, AES/ICP–MS) to biomedical research. Experimental approaches to studying the behavior and transformations of MNPs in vitro and in vivo are considered. The importance of proper sample preparation in simulating the behavior of nanoparticles in biological media is highlighted. We also examine the significance of preparation techniques for the accurate determination of dissolved and nanosized forms in biological samples. Lastly, we assess the potential for the comprehensive studies of MNP behavior within complex biological systems, pointing toward future directions in this dynamic and promising field of research.

Bilayer-Coating Strategy for Hydrophobic Nanoparticles Providing Colloidal Stability, Functionality, and Surface Protection in Biological Media.

The surface chemistry of nanoparticles is a key step on the pathway from particle design towards applications in biologically relevant environments. Here, a bilayer-based strategy for the surface modification of hydrophobic nanoparticles is introduced that leads to excellent colloidal stability in aqueous environments and good protection against disintegration, while permitting surface functionalization via simple carbodiimide chemistry. We have demonstrated the excellent potential of this strategy using upconversion nanoparticles (UCNPs), initially coated with oleate and therefore dispersible only in organic solvents. The hydrophobic oleate capping is maintained and a bilayer is formed upon addition of excess oleate. The bilayer approach renders protection towards luminescence loss by water quenching, while the incorporation of additional molecules containing amino functions yields colloidal stability and facilitates the introduction of functionality. The biological relevance of the approach was confirmed with the use of two model dyes, a photosensitizer and a nitric oxide (NO) probe that, when attached to the surface of the UCNPs, retained their functionality to produce singlet oxygen and detect intracellular NO, respectively. We present a simple and fast strategy to protect and functionalize inorganic nanoparticles in biological media, which is important for controlled surface engineering of nanosized materials for theranostic applications.

Strategie zur Beschichtung von hydrophoben Nanopartikeln mit einer Doppelschicht für kolloidale Stabilität, Funktionalität und Oberflächenschutz in biologischen Medien

AbstractDie Oberflächenchemie von Nanopartikeln ist ein wichtiger Schritt auf dem Weg vom Partikeldesign zur Anwendung in biologisch relevanten Umgebungen. Hier wird eine auf einer Doppelschicht (Bilayer) basierende Strategie für die Oberflächenmodifikation von hydrophoben Nanopartikeln vorgestellt, die diesen eine ausgezeichnete kolloidale Stabilität in wässriger Umgebung und einen guten Schutz gegen Auflösung verleiht und gleichzeitig eine Oberflächenfunktionalisierung durch einfache Carbodiimid‐Kopplungschemie ermöglicht. Wir haben das herausragende Potenzial dieser Strategie anhand von Upconversion‐Nanopartikeln (UCNPs) demonstriert, die ursprünglich mit Oleat beschichtet und daher nur in organischen Lösungsmitteln dispergierbar waren. Die hydrophobe Oleatbeschichtung bleibt dabei erhalten und bildet nach Zugabe von weiterem Oleat eine Doppelschicht. Dieser Ansatz schützt vor Lumineszenzlöschung durch Wassermoleküle, während der Einbau zusätzlicher Moleküle mit Aminogruppen in die Doppelschicht deren kolloidale Stabilität erhöht und die Funktionalisierbarkeit erleichtert. Die biologische Relevanz des Ansatzes wurde durch die Verwendung von zwei Modellfarbstoffen gezeigt, einem Photosensibilisator und einer Stickoxid‐ (NO−) Sonde, die, wenn sie an die Oberfläche der UCNPs gebunden sind, ihre Funktionalität zur Erzeugung von Singulett‐Sauerstoff bzw. zum Nachweis von intrazellulärem NO beibehalten. Wir stellen eine einfache und schnelle Methode zum Schutz und zur Funktionalisierung von anorganischen Nanopartikeln in biologischen Medien vor, die für die kontrollierte Oberflächengestaltung von Nanomaterialien für theranostische Anwendungen wichtig ist.

Colloidal Perspective on Targeted Drug Delivery to the Central Nervous System.

This article describes a new approach in targeted drug delivery to the central nervous system (CNS) in a significant departure from the predominant systematic drug administration attempting to penetrate the blood-brain barrier (BBB). Nanoparticles chemically conjugated to neural tract tracer proteins are capable of path-specific axonal retrograde transport, transneuronal transport, and anatomical tract flow to bypass the BBB. To celebrate the work by Dr. Bettye Washington Greene on the physical chemistry of colloidal particles, this article focuses on the physiochemical characteristics of the nanoparticles, various colloidal forces that impact the colloidal stability of nanoparticles in biological media, and surface chemistry strategies to avoid nanoparticle aggregation-induced poor therapeutic outcomes. The biological environment for the anatomical retrograde transport of neural tract tracers is examined to directly link factors impacting the colloidal stability of the new class of CNS-targeting nanoconjugates such as nanoconjugate size, shape, surface charge, surface chemistry, ionic strength, pH, and protein adsorption on the nanoparticle. We conclude with opportunities and challenges for future research.

Red-Absorbing Ru(II) Polypyridyl Complexes with Biotin Targeting Spontaneously Assemble into Nanoparticles in Biological Media.

Four new ruthenium(II) polypyridyl complexes were synthesized to study the effect of poly(ethylene glycol) and/or biotin conjugation on their physical and biological properties, including their hydrophilicity, their cellular uptake, and their phototoxicity. Unexpectedly, these complexes self-assembled into nanoparticles upon dilution in biological media. This behavior leads to their accumulation in lysosomes following their internalization by cells. While a significant increase in cellular uptake was observed for the biotin-conjugated complexes, it did not result in an increase in their phototoxicity. However, their high phototoxicity upon irradiation at long wavelengths (645-670 nm) and their self-assembling behavior make them a promising backbone for the development of new lysosome-targeted photosensitizers for photodynamic therapy.

Patterning of Self-Assembled Monolayers of Amphiphilic Multisegment Ligands on Nanoparticles and Design Parameters for Protein Interactions

Functionalization of nanoparticles with specific ligands is helpful to control specific diagnostic and therapeutic responses such as protein adsorption, cell targeting, and circulation. Precision delivery critically depends on a fundamental understanding of the interplay between surface chemistry, ligand dynamics, and interaction with the biochemical environment. Due to limited atomic-scale insights into the structure and dynamics of nanoparticle-bound ligands from experiments, relationships of grafting density and ligand chemistry to observable properties such as hydrophilicity and protein interactions remain largely unknown. In this work, we uncover how self-assembled monolayers (SAMs) composed of multisegment ligands such as thioalkyl-PEG-(N-alkyl)amides on gold nanoparticles can mimic mixed hydrophobic and hydrophilic ligand coatings, including control of patterns, hydrophilicity, and specific recognition properties. Our results are derived from molecular dynamics simulations with the INTERFACE-CHARMM36 force field at picometer resolution and comparisons to experiments. Small changes in ligand hydrophobicity, via adjusting the length of the N-terminal alkyl groups, tune water penetration by multiples and control superficial ordering of alkyl chains from 0 to 70% regularity. Further parameters include the grafting density of the ligands, curvature of the nanoparticle surfaces, type of solvent, and overall ligand length, which were examined in detail. We explain the thermodynamic origin of the formation of heterogeneous patterns of multisegment ligand SAMs and illustrate how different degrees of ligand order on the nanoparticle surface affect interactions with bovine serum albumin. The resulting design principles can be applied to a variety of ligand chemistries to customize the behavior of functionalized nanoparticles in biological media and enhance therapeutic efficiency.

Elucidating the Stability of Single-Chain Polymeric Nanoparticles in Biological Media and Living Cells.

The controlled folding of synthetic polymer chains into single-chain polymeric nanoparticles (SCPNs) of defined size and shape in water is a viable way to create compartmentalized, nanometer-sized structures for a range of biological applications. Understanding the relationship between the polymer’s microstructure and the stability of folded structures is crucial to achieving desired applications. Here, we introduce the solvatochromic dye Nile red into SCPNs and apply a combination of spectroscopic and microscopic techniques to relate polymer microstructure to nanoparticle stability in complex biological media and cellular environments. Our experimental data show that the polymer’s microstructure has little effect on the stability of SCPNs in biological media and cytoplasm of living cells, but only SCPNs comprising supramolecular benzene-1,3,5-tricarboxamide (BTA) motifs showed good stability in lysosomes. The results indicate that the polymer’s microstructure is vital to ensure nanoparticle stability in highly competitive environments: both hydrophobic collapse and a structured interior are required. Our study provides an accessible way of probing the stability of SCPNs in cellular environments and paves the way for designing highly stable SCPNs for efficient bio-orthogonal catalysis and sensing applications.

Biodegradation of metal-based ultra-small nanoparticles: A combined approach using TDA-ICP-MS and CE-ICP-MS

The knowledge of the fate of metal-containing nanoparticles in biological media in aqueous media is of utmost importance for the future use of these promising theranostic agents for clinical applications. A methodology based on the combination of TDA-ICP-MS and CE-ICP-MS was applied to study the degradation pathway of AGuIX, a phase 2 clinical ultrasmall gadolinium-containing nanoparticle. Nanoparticle size measurements and gadolinium speciation performed in different media (phosphate buffer, urine and serum) demonstrated an accelerated dissolution of AGuIX in serum, without any release of free gadolinium for each medium.

Role of Aqueous-Phase Calcination in Synthesis of Ultra-Stable Dye-Embedded Fluorescent Nanoparticles for Cellular Probing.

Fluorescence imaging is a major driver of discovery in biology, and an invaluable asset in clinical diagnostics. To overcome quenching limitations of conventional fluorescent dyes and further improve intensity, nanoparticle-based constructs have been the subject of intense investigation, and within this realm, dye-doped silica-coated nanoparticles have garnered significant attention. Despite their growing popularity in research, fluorescent silica nanoparticles suffer from a significant flaw. The degradation of these nanoparticles in biological media by hydrolytic dissolution is underreported, leading to serious misinterpretations, and limiting their applicability for livecellandinvivoimaging. Here, the development of an ultra-stable, dye-embedded, silica-coated metal nanoparticle is reported, and its superior performance in long-term live cell imaging is demonstrated. While conventional dye-doped silica nanoparticles begin to degrade within an hour in aqueous media, by leveraging a modified liquid calcination process, this new construct is shown to be stableforat least 24 h.The stability of this metal-enhanced fluorescent probe in biologically relevant temperatures and media, and its demonstrated utility for cell imaging, paves the way for its future adoption in biomedical research.

Correlation between in vitro stability and pharmacokinetics of poly(ε-caprolactone)-based micelles loaded with a photosensitizer

Polymeric micelles are extensively investigated as drug delivery systems for hydrophobic drugs including photosensitizers (PSs). In order to benefit from micelles as targeted delivery systems for PS, rather than only solubilizers, the stability and cargo retention of the (PS-loaded) micelles should be properly assessed in biologically relevant media to get insight into the essential parameters predicting their in vivo performance (i.e., pharmacokinetics). In the present study, asymmetric flow field-flow fractionation (AF4) was used to investigate the in vitro stability in human plasma of empty and meta-tetra(hydroxyphenyl)chlorin (mTHPC)-loaded dithiolane-crosslinked micelles based on poly(ɛ-caprolactone)-co-poly(1,2-dithiolane‑carbonate)-b-poly(ethylene glycol) (p(CL-co-DTC)-PEG) and non (covalently)-crosslinked micelles composed of poly(ε-caprolactone)-b-poly(ethylene glycol) (pCL-PEG). AF4 allows separation of the micelles from plasma proteins, which showed that small non (covalently)-crosslinked pCL9-PEG (17 nm) and pCL15-PEG (22 nm) micelles had lower stability in plasma than pCL23-PEG micelles with larger size (43 nm) and higher degree of crystallinity of pCL, and had also lower stability than covalently crosslinked p(CL9-DTC3.9)-PEG and p(CL18-DTC7.5)-PEG micelles with similar small sizes (~20 nm). In addition, PS (re)distribution to specific plasma proteins was observed by AF4, giving strong indications for the (in)stability of PS-loaded micelles in plasma. Nevertheless, fluorescence spectroscopy in human plasma showed that the retention of mTHPC in non (covalently)-crosslinked but semi-crystalline pCL23-PEG micelles (>8 h) was much longer than that in covalently crosslinked p(CL18-DTC7.5)-PEG micelles (~4 h). In line with this, in vivo circulation kinetics showed that pCL23-PEG micelles loaded with mTHPC had significantly longer half-life values (t½-β of micelles and mTHPC was 14 and 18 h, respectively) than covalently crosslinked p(CL18-DTC7.5)-PEG micelles (t½-β of both micelles and mTHPC was ~2 h). As a consequence, long circulating pCL23-PEG micelles resulted in significantly higher tumor accumulation of both the micelles and loaded mTHPC as compared to short circulating p(CL18-DTC7.5)-PEG micelles. These in vivo data were in good agreement with the in vitro stability studies.In conclusion, the present study points out that AF4 and fluorescence spectroscopy are excellent tools to evaluate the (in)stability of nanoparticles in biological media and thus predict the (in)stability of drug loaded nanoparticles after i.v. administration, which is favorable to screen promising delivery systems with reduced experimental time and costs and without excessive use of animals.

Colloidal Stability and Redispersibility of Mesoporous Silica Nanoparticles in Biological Media.

The outreach of nanoparticle-based medical treatments has been severely hampered due to the imbalance between the efforts in designing extremely complex materials and the general lack of studies devoted to understanding their colloidal stability in biological environments. Over the years, the scientific community has neglected the relevance related to the nanoparticles' colloidal state, which consequently resulted in very poor bench-to-clinic translation. In this work, we show how mesoporous silica nanoparticles (MSNs, one of the most promising and tested drug delivery platforms) can be efficiently synthesized and prepared, resulting in a colloidally stable system. We first compared three distinct methods of template removal of MSNs and evaluated their ultimate colloidal stability. Then, we also proposed a simple way to prevent aggregation during the drying step by adsorbing BSA onto MSNs. The surface modification resulted in colloidally stable particles that are successfully redispersed in biologically relevant medium while retaining high hemocompatibility and low cytotoxicity.

Comprehensive study of stability of copper oxide nanoparticles in complex biological media

In response to the rapid development of nanomaterials in the biomedical field, especially in advanced cancer therapy, more comprehensive studies regarding the safety of nanomaterials are continuously required. Currently, several fundamental studies concerning the fate of nanoparticles in the biological media are needed, especially before in vivo studies. Inspired by these approaches, and the wide variation on result in the literature regarding copper (I) oxide nanoparticles (Cu2O NPs), in this article we modified sulfobetaine-stabilzed Cu2O NPs with glutathione (GSH) and hyaluronic acid (HA) tumor targeting ligands to investigate the effects of nanoparticles design on their stability in four different biological fluids: simulated saliva, human blood plasma, gastric and intestinal fluids. In addition, we examined the kinetics of copper ions release under physiological conditions, and according to mathematical models, we proposed the most probable mechanism of mass transport. The GSH- and HA-modified, as well as unmodified sulfobetaine-stabilized Cu2O NPs suspended in all tested biological fluids, formed agglomerates with sizes varying from 150 to 700 nm. The highest copper ions content was observed for Cu2O NPs in simulated gastric fluid. The formation of aggregates and copper ions release in contact with biological fluids influenced the Cu2O NPs cytotoxicity. The results indicated that cancer cells were more susceptible to HA-modified copper oxide NPs action.

Interaction of gold nanorods with cell culture media: Colloidal stability, cytotoxicity and cellular death modality

Behavior of nanoparticles in biological media is crucial in predicting their cellular responses. In this study, gold nanorods (GNR) were chosen as a model to evaluate the influence of nanoparticle's surface modification and the composition of cell culture media on the colloidal stability and dispersibility of the nanoparticles in media. Optical spectra, zeta potential and hydrodynamic size were measured to evaluate the colloidal stability. The results indicate that incorporation of fetal bovine serum (FBS) into the cell culture media drastically enhanced their colloidal stability by protein adsorption, while nanoparticles dispersed in serum-free media demonstrated obvious aggregation. Nanoparticles dispersed in serum-free media showed high rate and extent of cellular internalization and dose-independent cytotoxicity compared to those dispersed in serum-containing media. Flow cytometry revealed that both late apoptosis and necrosis are responsible for the cellular death of breast cancer cells exposed to well-dispersed GNR in serum-containing media, while cells exposed to nano-aggregates experienced necrosis as the major modality of their cellular death.

Surface Coating Structure and Its Interaction with Cytochrome c in EG6-Coated Nanoparticles Varies with Surface Curvature.

The composition, orientation, and conformation of proteins in biomolecular coronas acquired by nanoparticles in biological media contribute to how they are identified by a cell. While numerous studies have investigated protein composition in biomolecular coronas, relatively little detail is known about how the nanoparticle surface influences the orientation and conformation of the proteins associated with them. We previously showed that the peripheral membrane protein cytochrome c adopts preferred poses relative to negatively charged 3-mercaptopropionic acid (MPA)-gold nanoparticles (AuNPs). Here, we employ molecular dynamics simulations and complementary experiments to establish that cytochrome c also assumes preferred poses upon association with nanoparticles functionalized with an uncharged ligand, specifically ω-(1-mercaptounde-11-cyl)hexa(ethylene glycol) (EG6). We find that the display of the EG6 ligands is sensitive to the curvature of the surface-and, consequently, the effective diameter of the nearly spherical nanoparticle core-which in turn affects the preferred poses of cytochrome c.

Influence of Biocorona Formation on the Transformation and Dissolution of Cobalt Nanoparticles under Physiological Conditions.

Cobalt (Co) nanoparticles (NPs) are produced in different applications and unintentionally generated at several occupational and traffic settings. Their diffuse dispersion may lead to interactions with humans and aquatic organisms via different exposure routes that include their transformation/dissolution in biological media. This paper has investigated the particle stability and reactivity of Co NPs (dispersed by sonication prior to exposure) interacting with selected individual biomolecules (amino acids, polypeptides, and proteins) in phosphate-buffered saline (PBS). No or minor adsorption of amino acids (glutamine, glutamic acid, lysine, and cysteine) was observed on the Co NPs, independent of the functional group and charge. Instead, phosphate adsorption resulted in the formation of a surface layer (a corona) of Co phosphate. The adsorption of larger biomolecules (polyglutamic acid, polylysine, lysozyme, and mucin) was evident in parallel with the formation of Co phosphate. The dissolution of the Co NPs was rapid as 35–55% of the particle mass was dissolved within the first hour of exposure. The larger biomolecules suppressed the dissolution initially compared to exposure in PBS only, whereas the dissolution was essentially unaffected by the presence of amino acids, with cysteine as an exception. The formation of Co phosphate on the NP surface reduced the protective properties of the surface oxide of the Co NPs, as seen from the increased levels of the released Co when compared with the nonphosphate-containing saline. The results underline the diversity of possible outcomes with respect to surface characteristics and dissolution of Co NPs in biological media and emphasize the importance of surface interactions with phosphate on the NP characteristics and reactivity.

Gram‐Negative Bacteria Targeting Mediated by Carbohydrate–Carbohydrate Interactions Induced by Surface‐Modified Nanoparticles

AbstractAntibiotic resistant pathogens are a modern threat to the human health. As a worldwide spreading problem, there is an urgency for new strategies to minimize antibiotic resistance, particularly the super‐resistant strains. Here, the efficient design of carbohydrate‐coated silica nanoparticles is reported which specifically target Gram‐negative bacteria cells. The system is functionalized with gluconamide moieties and demonstrates increased binding ability to the bacterial membrane, enabling controlled drug delivery onto the pathogen wall. In addition, the high stability of the nanoparticles in biological media and the lack of non‐specific protein adhesion are engendered by such functionalization, which also demonstrates low cytotoxicity and hemolytic activity prevention. Local interaction between nanoparticles and the bacterium membrane is experimentally accessed at the biomolecular level unveiling a short‐range chemical connection. Atomistic molecular dynamics simulations depict the rapid penetration of gluconamide in the lipopolysaccharide region of the bacterial outer membrane, corroborating the experimental findings. Thus, this novel outer membrane‐targeting platform provides a new strategy to reduce drug intake and, hence, minimize bacterial resistance.

Highly Crystalline WO3 Nanoparticles Are Nontoxic to Stem Cells and Cancer Cells

Tungsten oxide sol, containing highly crystalline nanoparticles of orthorhombic WO3 and having good sedimentation stability, was synthesized using a facile, ultrasonic-assisted technique. An additional steric stabilizer, dextran, was proposed to enhance the stability of WO3 nanoparticles in biological media and to reduce their in vivo toxicity. The cytotoxicity of dextran-stabilized and nonstabilized WO3 sols was studied in vitro using dental pulp stem (DPS) cell lines and breast cancer (MCF-7) cell lines. Both tungsten oxide sols demonstrated low cytotoxicity and low genotoxicity for both stem cells and malignant cells and only slightly reduced their metabolic activity in the concentration range studied (from 0.2 to 200 μg/ml). The data obtained support possible theranostic applications of tungsten oxide colloidal solutions.