Monica Carril

Gold-Based Nanomaterials for Applications in Nanomedicine

In this review, an overview of the current state-of-the-art of gold-based nanomaterials (Au NPs) in medical applications is given. The unique properties of Au NPs, such as their tunable size, shape, and surface characteristics, optical properties, biocompatibility, low cytotoxicity, high stability, and multifunctionality potential, among others, make them highly attractive in many aspects of medicine. First, the preparation methods for various Au NPs including functionalization strategies for selective targeting are summarized. Second, recent progresses on their applications, ranging from the diagnostics to therapeutics are highlighted. Finally, the rapidly growing and promising field of gold-based theranostic nano-platforms is discussed. Considering the great body of existing information and the high speed of its renewal, we chose in this review to generalize the data that have been accumulated during the past few years for the most promising directions in the use of Au NPs in current medical research.

Techniques for the experimental investigation of the protein corona

Due to its enormous relevance the corona formation of adsorbed proteins around nanoparticles is widely investigated. A comparison of different experimental techniques is given. Direct measurements of proteins, such as typically performed with mass spectrometry, will be compared with indirect analysis, in which instead information about the protein corona is gathered from changes in the properties of the nanoparticles. The type of measurement determines also whether before analysis purification from unbound excess proteins is necessary, which may change the equilibrium, or if measurements can be performed in situ without required purification. Pros and contras of the different methods will be discussed.

Targeted gold-coated iron oxide nanoparticles for CD163 detection in atherosclerosis by MRI

CD163 is a membrane receptor expressed by macrophage lineage. Studies performed in atherosclerosis have shown that CD163 expression is increased at inflammatory sites, pointing at the presence of intraplaque hemorrhagic sites or asymptomatic plaques. Hence, imaging of CD163 expressing macrophages is an interesting strategy in order to detect atherosclerotic plaques. We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI. Firstly, the specificity of the targeted probe was validated in vitro by incubation of the probe with CD163(+) or (−) macrophages. The probe was able to selectively detect CD163(+) macrophages both in human and murine cells. Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI. The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection. Hence, we have developed a highly sensitive targeted probe capable of detecting CD163-expressing macrophages that could provide useful information about the state of the atheromatous lesions.

CD163-Macrophages Are Involved in Rhabdomyolysis-Induced Kidney Injury and May Be Detected by MRI with Targeted Gold-Coated Iron Oxide Nanoparticles.

Macrophages play an important role in rhabdomyolysis-acute kidney injury (AKI), although the molecular mechanisms involved in macrophage differentiation are poorly understood. We analyzed the expression and regulation of CD163, a membrane receptor mainly expressed by anti-inflammatory M2 macrophages, in rhabdomyolysis-AKI and developed targeted probes for its specific detection in vivo by MRI. Intramuscular injection of glycerol in mice promoted an early inflammatory response, with elevated proportion of M1 macrophages, and partial differentiation towards a M2 phenotype in later stages, where increased CD163 expression was observed. Immunohistological studies confirmed the presence of CD163-macrophages in human rhabdomyolysis-AKI. In cultured macrophages, myoglobin upregulated CD163 expression via HO-1/IL-10 axis. Moreover, we developed gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody that specifically targeted CD163 in kidneys from glycerol-injected mice, as determined by MRI studies, and confirmed by electron microscopy and immunological analysis. Our findings are the first to demonstrate that CD163 is present in both human and experimental rhabdomyolysis-induced AKI, suggesting an important role of this molecule in this pathological condition. Therefore, the use of probes targeting CD163-macrophages by MRI may provide important information about the cellular composition of renal lesion in rhabdomyolysis.

Activatable probes for diagnosis and biomarker detection by MRI

Magnetic resonance imaging (MRI) is a non-invasive imaging technique with widespread use in diagnosis. Frequently, contrast in MRI is enhanced with the aid of a contrast agent, among which smart, responsive, OFF/ON or activatable probes are of particular interest. These kinds of probes elicit a response to selective stimuli, evidencing the presence of enzymes or acidic pH, for instance. In this review, we will focus on smart probes that are detectable by both 1H and 19F MRI, frequently based on nanomaterials. We will discuss the triggering factors and the strategies employed thus far to activate each probe.

In situ detection of the protein corona in complex environments

Colloidal nanoparticles (NPs) are a versatile potential platform for in vivo nanomedicine. Inside blood circulation, NPs may undergo drastic changes, such as by formation of a protein corona. The in vivo corona cannot be completely emulated by the corona formed in blood. Thus, in situ detection in complex media, and ultimately in vivo, is required. Here we present a methodology for determining protein corona formation in complex media. NPs are labeled with 19F and their diffusion coefficient measured using 19F diffusion-ordered nuclear magnetic resonance (NMR) spectroscopy. 19F diffusion NMR measurements of hydrodynamic radii allow for in situ characterization of NPs in complex environments by quantification of protein adsorption to the surface of NPs, as determined by increase in hydrodynamic radius. The methodology is not optics based, and thus can be used in turbid environments, as in the presence of cells.In situ detection of protein coronas is usually performed via optical methods, but light scattering may hamper these measurements. Here, the authors use diffusion NMR techniques to characterize protein corona formation on 19F-labeled nanoparticles in blood and other complex media.

Multimodal 19F NMR Dopamine Detection and Imaging with a Nanoparticle‐Based Displacement Assay

Nanoconjugates composed of gold nanoparticles (core diameter=1.9 nm) coated with thioundecyl-d-glucopyranosides and fluorinated phenylboronic acids can detect diol-containing derivatives by means of 19 F NMR spectroscopic analysis. The spectra of nanoconjugate solutions display broad signals due to the fast relaxation of the 19 F nuclei caused by nanoparticle grafting. When dopamine is added, the formation of a boronate ester between the analyte and the fluorinated boronic acid causes the release of the latter in solution and consequent sharpening of the NMR signals. Dopamine can be selectively detected through magnetic resonance imaging (MRI) and NMR spectroscopic analysis with respect to glucose and galactose with a detection limit of 20 μm. The chemical shift of the released ester is diagnostic of the recognized analyte. Consequently, the sensor also enables the simultaneous detection of different analytes.

Study of Fluorinated Quantum Dots-Protein Interactions at the Oil/Water Interface by Interfacial Surface Tension Changes

Understanding the interaction of nanoparticles with proteins and how this interaction modifies the nanoparticles’ surface is crucial before their use for biomedical applications. Since fluorinated materials are emerging as potential imaging probes and delivery vehicles, their interaction with proteins of biological interest must be studied in order to be able to predict their performance in real scenarios. It is known that fluorinated planar surfaces may repel the unspecific adsorption of proteins but little is known regarding the same process on fluorinated nanoparticles due to the scarce examples in the literature. In this context, the aim of this work is to propose a simple and fast methodology to study fluorinated nanoparticle-protein interactions based on interfacial surface tension (IFT) measurements. This technique is particularly interesting for fluorinated nanoparticles due to their increased hydrophobicity. Our study is based on the determination of IFT variations due to the interaction of quantum dots of ca. 5 nm inorganic core/shell diameter coated with fluorinated ligands (QD_F) with several proteins at the oil/water interface. Based on the results, we conclude that the presence of QD_F do not disrupt protein spontaneous film formation at the oil/water interface. Even if at very low concentrations of proteins the film formation in the presence of QD_F shows a slower rate, the final interfacial tension reached is similar to that obtained in the absence of QD_F. The differential behaviour of the studied proteins (bovine serum albumin, fibrinogen and apotransferrin) has been discussed on the basis of the adsorption affinity of each protein towards DCM/water interface and their different sizes. Additionally, it has been clearly demonstrated that the proposed methodology can serve as a complementary technique to other reported direct and indirect methods for the evaluation of nanoparticle-protein interactions at low protein concentrations.