Adriano Boni

Surface functionalisation regulates polyamidoamine dendrimer toxicity on blood–brain barrier cells and the modulation of key inflammatory receptors on microglia

Abstract Dendrimers are branched polymers with spherical morphology. Their tuneable chemistry and surface modification make them valuable nanomaterials for biomedical applications. In view of possible dendrimer uses as brain-aimed nanocarriers, the authors studied amine- and lipid-functionalised (G4) polyamidoamine (PAMAM) biocompatibility with cell population forming the blood–brain barrier (BBB). Both amine-PAMAM and lipid-PAMAM dendrimers were able to enter endothelial and primary neural cells. However, only amine-PAMAM damaged cell membranes in a dose-dependent manner. Transmission electron microscopy evidenced the ability of dendrimers to precipitate salts and serum components present in culture medium that slightly increased toxicity of the amine-PAMAM. Amine- and lipid-PAMAM were both able to cross the BBB and differently induced CD11b and CCR2 overexpression on primary CX3CR1-GFP murine microglia in vitro. These data emphasise the role of dendrimer surface functionalisation in toxicity and neural immune cell activation, raising concerns about possible neuroinflammatory reactions.

Design and characterization of antimicrobial usnic acid loaded-core/shell magnetic nanoparticles

The application of magnetic nanoparticles (MNPs) in medicine is considered much promising especially because they can be handled and directed to specific body sites by external magnetic fields. MNPs have been investigated in magnetic resonance imaging, hyperthermia and drug targeting. In this study, properly functionalized core/shell MNPs with antimicrobial properties were developed to be used for the prevention and treatment of medical device-related infections. Particularly, surface-engineered manganese iron oxide MNPs, produced by a micro-emulsion method, were coated with two different polymers and loaded with usnic acid (UA), a dibenzofuran natural extract possessing antimicrobial activity. Between the two polymer coatings, the one based on an intrinsically antimicrobial cationic polyacrylamide (pAcDED) resulted to be able to provide MNPs with proper magnetic properties and basic groups for UA loading. Thanks to the establishment of acid-base interactions, pAcDED-coated MNPs were able to load and release significant drug amounts resulting in good antimicrobial properties versus Staphylococcus epidermidis (MIC = 0.1 mg/mL). The use of pAcDED having intrinsic antimicrobial activity as MNP coating in combination with UA likely contributed to obtain an enhanced antimicrobial effect. The developed drug-loaded MNPs could be injected in the patient soon after device implantation to prevent biofilm formation, or, later, in presence of signs of infection to treat the biofilm grown on the device surfaces.

Water dispersal and functionalization of hydrophobic iron oxide nanoparticles with lipid-modified poly(amidoamine) dendrimers.

A novel and facile method for water dispersal of hydrophobic iron oxide nanoparticles based on the amphiphilic PAMAM-C12 dendrimer is described. Stable and highly concentrated water dispersions of multifunctional magnetic nanoparticles were obtained with this single-step approach, and showed interesting relaxometric properties for MRI applications. Importantly, this method does not require substitution of the native hydrophobic capping under nonmild reaction conditions, thus preserving the structural and magnetic properties of the nanoparticles, and extending the possibility of conjugation with thermally labile groups.

USPIO-loaded red blood cells as a biomimetic MR contrast agent: a relaxometric study.

Red blood cells (RBCs) loaded with iron oxide nanoparticles have been proposed as biomimetic constructs with long half-life (ca. 20 days) in the blood compartment and potentially interesting properties (such as relaxivity) as intravascular contrast agents for magnetic resonance imaging. However, the encapsulation of nanoparticles into RBCs might affect their magnetic properties and relaxivity, which may be significantly different from the native suspension. Here, we present a relaxometric study of P904, a novel ultra small iron oxide nanoparticle developed by Guerbet, enclosed in human RBCs. We measured longitudinal (r1 ) and transverse (r2 ) relaxivity over a wide range of Larmor frequencies (0.01-300 MHz) in samples of P904-loaded RBCs, and in control samples with P904 nanoparticles dispersed in blood. Internalization of P904 into RBCs resulted in smaller r1 , and in a very high r2 /r1 ratio (232) at the highest field. Moreover, a shift of the Curie peak to high fields was observed in P904-loaded RBCs, possibly the result of nanoparticle size selection caused by the internalization process. High r2 relaxivity together with a high r2 /r1 ratio and a very long blood half-life make P904-loaded RBCs a promising blood-pool negative contrast agent for MR diagnostic applications.

Gd-doped BNNTs as T2-weighted MRI contrast agents.

This work describes, for the first time, doping of boron nitride nanotubes (BNNTs) with gadolinium (Gd@BNNTs), a stable functionalization that permits non-invasive BNNT tracking via magnetic resonance imaging (MRI). We report the structure, Gd loading, and relaxometric properties in water suspension at 7 T of Gd@BNNTs, and show the behaviour of these nanostructures as promising T2-weighted contrast agents. Finally, we demonstrate their complete biocompatibility in vitro on human neuroblastoma cells, together with their ability to effectively label and affect contrast in MRI images at 7 T.

Ultrasmall superparamagnetic iron oxide nanoparticles with titanium-N,N-dialkylcarbamato coating

This work deals with the preparation and physical-chemical characterization of new ultrasmall iron oxide superparamagnetic nanoparticles (USPIONs) functionalized with titanium-N,N-dialkylcarbamato. The preparation was performed starting with monodispersed USPIONs covered with oleic acid, synthesized by thermal-decomposition, and subsequently functionalized with metal-carbamato by a ligand-exchange reaction. The surface and coating structure was characterized by infrared (FT-IR) spectroscopy on the solid powders and thermogravimetry (TG) coupled with an FT-IR detector in order to better investigate the self-assembling properties of the coating. A detailed dimensional and morphological study was carried out by transmission electron microscopy (TEM) and atomic force microscopy (AFM) analysis. Zero-field-cooled (ZFC) and field-cooled (FC) magnetic susceptibility curves as well as the magnetization behavior as a function of temperature were investigated on both the starting oleic-USPIONs and those covered by titanium-N,N-dialkylcarbamato. These results confirmed the superparamagnetic properties of the new nanoparticles (NPs), highlighting the quite high saturation value of the magnetization. Based on the results obtained by combining different experimental techniques, a model of the coating structure and ligand organization around the magnetic core is proposed for both NPs, i.e. the starting USPIONs covered by oleic acid and the new USPIONs functionalized by titanium-N,N-dialkylcarbamato.

Water-Dispersible Three-Dimensional LC-Nanoresonators

Nanolithography techniques enable the fabrication of complex nanodevices that can be used for biosensing purposes. However, these devices are normally supported by a substrate and their use is limited to in vitro applications. Following a top-down procedure, we designed and fabricated composite inductance-capacitance (LC) nanoresonators that can be detached from their substrate and dispersed in water. The multimaterial composition of these resonators makes it possible to differentially functionalize different parts of the device to obtain stable aqueous suspensions and multi-sensing capabilities. For the first time, we demonstrate detection of these devices in an aqueous environment, and we show that they can be sensitized to their local environment and to chemical binding of specific molecular moieties. The possibility to optically probe the nanoresonator resonance in liquid dispersions paves the way to a variety of new applications, including injection into living organisms for in vivo sensing and imaging.

Lipid-modified dendrimers as a tool for the design of nanoparticle-based multimodal MRI contrast agents

A fast and easy method for water transfer of iron oxide nanoparticle based on the hydrophobic interaction of oleic acid coated nanoparticles with the amphiphilic PAMAM-C12 dendrimer is described. The process may be conveniently performed in water and yields nanoparticles with good size distribution, diameter modulation and high crystallinity. The nanoparticles have been functionalized with gadolinium ions to obtain T1/T2 dual mode contrast agents. Furthermore, the possibility to deliver and release lipophilic drugs was investigated.