Elisa Cabrini

A Molecular Thermometer for Nanoparticles for Optical Hyperthermia

We developed an all-optical method to measure the temperature on gold (nanorods and nanostars) and magnetite nanoparticles under near-infrared and radiofrequency excitation by monitoring the excited state lifetime of Rhodamine B that lies within =/~20 nm from the nanoparticle surface. We reached high temperature sensitivity (0.029 ± 0.001 ns/°C) and low uncertainty (±0.3 °C). Gold nanostars are =/~3 and =/~100 times more efficient than gold nanorods and magnetite nanoparticles in inducing localized hyperthermia.

Thermal and Chemical Stability of Thiol Bonding on Gold Nanostars.

The stability of thiol bonding on the surface of star-shaped gold nanoparticles was studied as a function of temperature in water and in a set of biologically relevant conditions. The stability was evaluated by monitoring the release of a model fluorescent dye, Bodipy-thiol (BDP-SH), from gold nanostars (GNSs) cocoated with poly(ethylene glycol) thiol (PEG-SH). The increase in the BDP-SH fluorescence emission, quenched when bound to the GNSs, was exploited to this purpose. A maximum 15% dye release in aqueous solution was found when the bulk temperature of gold nanostars solutions was increased to T = 42 °C, the maximum physiological temperature. This fraction reduces 3-5% for temperatures lower than 40 °C. Similar results were found when the temperature increase was obtained by laser excitation of the near-infrared (NIR) localized surface plasmon resonance of the GNSs, which are photothermally responsive. Besides the direct impact of temperature, an increased BDP-SH release was observed upon changing the chemical composition of the solvent from pure water to phosphate-buffered saline and culture media solutions. Moreover, also a significant fraction of PEG-SH was released from the GNS surface due to the increase in temperature. We monitored it with a different approach, that is, by using a coating of α-mercapto-ω-amino PEG labeled with tetramethylrhodamine isothiocyanate on the amino group, that after heating was separated from GNS by ultracentrifugation and the released PEG was determined by spectrofluorimetric techniques on the supernatant solution. These results suggest some specific limitations in the use of the gold-thiolate bond for coating of nanomaterials with organic compounds in biological environments. These limitations come from the duration and the intensity of the thermal treatment and from the medium composition and could also be exploited in biological media to modulate the in vivo release of drugs.

Fabrication of Inkjet-Printed Gold Nanostar Patterns with Photothermal Properties on Paper Substrate

Inkjet printing technology has brought significant advances in patterning various functional materials that can meet important challenges in personalized medical treatments. Indeed, patterning of photothermal active anisotropic gold nanoparticles is particularly promising for the development of low-cost tools for localized photothermal therapy. In the present work, stable inks containing PEGylated gold nanostars (GNSs) were prepared and inkjet printed on a pigment-coated paper substrate. A significant photothermal effect (ΔT ≅ 20 °C) of the printed patterns was observed under near infrared (NIR) excitation of the localized surface plasmon resonance (LSPR) of the GNS with low laser intensity (I ≅ 0.2 W/cm(2)). Besides the pronounced photothermal effect, we also demonstrated, as an additional valuable effect, the release of a model fluorescent thiol-terminated Bodipy dye (BDP-SH) from the printed gold surface, both under bulk heating and NIR irradiation. These preliminary results suggest the way of the development of a new class of low-cost, disposable, and smart devices for localized thermal treatments combined with temperature-triggered drug release.

Gold nanostars coated with neutral and charged polyethylene glycols: A comparative study of in-vitro biocompatibility and of their interaction with SH-SY5Y neuroblastoma cells.

Gold nanostars (GNS) have been coated with four different polyethylene glycols (PEGs) equipped with a -SH function for grafting on the gold surface. These PEGs have different chain lengths with average MW=2000, 3000, 5000 and average number of -O-CH2-CH2 - units 44, 66, and 111, respectively. Two are neutral and two are terminated with -COOH and -NH2 functions, thus bearing negative and positive charges at physiological pH, thanks to the formation of carboxylate and ammonium groups. The negative charge of the GNS coated with PEG carboxylate has also been exploited to further coat the GNS with the PAH (polyallylamine hydrochloride) cationic polymer. Vitality tests have been carried out on SH-SY5Y cells treated with the five differently coated GNS for 4, 24, and 48 h, at Au concentrations ranging from 1.25 to 100 μg/mL. The same tests have been repeated with the pure PEGs and PAH. Excellent biocompatibility was found for all PEGs, independently on charge and chain length, both for coated GNS and for the pure polymers. On the contrary, poor biocompatibility was found for PAH overcoated GNS and for pure PAH, although the latter only at high concentrations. Exploiting the two-photon luminescence of GNS, we have found by confocal laser scanning microscopy that when GNS are coated with PEGs they do not enter SH-SY5Y cells, while when overcoated with PAH they massively penetrate into the cytoplasm. This causes cell death by dramatically changing cell morphology, as demonstrated also by atomic force microscopy.

Gold nanostar–polymer hybrids for siRNA delivery: Polymer design towards colloidal stability and in vitro studies on breast cancer cells

To overcome the low bioavailability of siRNA (small interfering RNA) and to improve their transfection efficiency, the use of non-viral delivery carriers is today a feasible approach to transform the discovery of these incredibly potent and versatile drugs into clinical practice. Polymer-modified gold nanoconstructs (AuNCs) are currently viewed as efficient and safe intracellular delivery carriers for siRNA, as they have the possibility to conjugate the ability to stably entrap and deliver siRNAs inside cells with the advantages of gold nanoparticles, which can act as theranostic agents and radiotherapy enhancers through laser-induced hyperthermia. In this study, AuNCs were prepared by coating Gold Nano Stars (GNS) with suitable functionalised polymers, to give new insight on the choice of the coating in order to obtain colloidal stability, satisfying in vitro transfection behaviour and reliability in terms of homogeneous results upon GNS type changing. For this goal, GNS synthesized with three different sizes and shapes were coated with two different polymers: i) α-mercapto-ω-amino polyethylene glycol 3000Da (SH-PEG3000-NH2), a hydrophilic linear polymer; ii) PHEA-PEG2000-EDA-LA (PPE-LA), an amphiphilic hydroxyethylaspartamide copolymer containing a PEG moiety. Both polymers contain SH or SS groups for anchoring on gold surface and NH2 groups, which can be protonated in order to obtain a positive surface for successive siRNA layering. The effect of the features of the coating polymers on siRNA layering, and the extent of intracellular uptake and luciferase gene silencing effect were evaluated for each of the obtained coated GNS. The results highlight that amphiphilic biocompatible polymers with multi-grafting function are more suitable for ensuring the colloidal stability and the effectiveness of these colloidal systems, compared to the coating with linear PEG.

Tunable coating of gold nanostars: tailoring robust SERS labels for cell imaging

Surface modification of noble metal nanoparticles with mixed molecular monolayers is one of the most powerful tools in nanotechnology, and is used to impart and tune new complex surface properties. In imaging techniques based on surface enhanced Raman spectroscopy (SERS), precise and controllable surface modifications are needed to carefully design reproducible, robust and adjustable SERS nanoprobes. We report here the attainment of SERS labels based on gold nanostars (GNSs) coated with a mixed monolayer composed of a poly ethylene glycol (PEG) thiol (neutral or negatively charged) that ensure stability in biological environments, and of a signalling unit 7-Mercapto-4-methylcoumarin as a Raman reporter molecule. The composition of the coating mixture is precisely controlled using an original method, allowing the modulation of the SERS intensity and ensuring overall nanoprobe stability. The further addition of a positively charged layer of poly (allylamine hydrocloride) on the surface of negatively charged SERS labels does not change the SERS response, but it promotes the penetration of GNSs in SH-SY5Y neuroblastoma cells. As an example of an application of such an approach, we demonstrate here the internalization of these new labels by means of visualization of cell morphology obtained with SERS mapping.

A bistren cryptand with a remote thioether function: Cu( ii ) complexation in solution and on the surface of gold nanostars

A new macrobicyclic ligand capable of binding two Cu2+ cations has been synthesized and its protonation and coordinative properties fully determined in aqueous solution. A thioether moiety was appended on the ligand backbone. This does not influence the ligand coordination ability but allows us to graft its bis-copper complex on the surface of a self-assembled monolayer of gold nanostars (GNS), in turn grafted on glass slides pre-functionalized with a layer of a silane-bearing polyethyleneimine polymer. The release of copper ions from the GNS monolayers was also investigated, finding a general agreement with the coordination properties of the complex in solution, although the bis-copper complex displays an increased kinetic inertness when grafted on the glass slides. The photothermal properties of the GNS monolayer were studied with and without the overlayer of the Cu2+ complex, finding no influence of the latter but disclosing that the bis-copper complex detachment is promoted by local T increase due to laser irradiation.

Photothermal effect of gold nanostars inkjet-printed on coated paper substrate under near-infrared irradiation

The research and development of personalized medical treatments is increasing steadily fostered by its large societal impact. The ability of non-spherical gold nanoparticles to locally and efficiently release heat when irradiated in Near Infrared (NIR) wavelength region is a promising tool for photothermal medical therapies. In the present work, stable inks containing PEGylated gold nanostars (GNS) were obtained and inkjet-printed on a pigment coated paper substrate. Significant photothermal effect of the printed patterns was observed under Near Infrared (NIR) excitation of the Localized Surface Plasmon Resonance (LSPR) of the GNS. These preliminary results support, in perspective, the application of printed GNS patterns for thermal medical treatments either by direct localized heating, or by temperature triggered drug release.

Photothermal effect of gold nanostar patterns inkjet-printed on coated paper substrates with different permeability

Summary Inkjet printing of spherical gold nanoparticles is widely applied in the fabrication of analytical and diagnostics tools. These methods could be extended to non-spherical gold nanoparticles that can efficiently release heat locally when irradiated in the near infrared (NIR) wavelength region, due to localized surface plasmon resonance (LSPR). However, this promising application requires the ability to maintain high efficiency and tunability of the NIR LSPR of the printed nanoparticles. In this study stable inks containing PEGylated gold nanostars (GNS) were fabricated and successfully inkjet-printed onto differently coated paper substrates with different porosity and permeability. A pronounced photothermal effect was observed under NIR excitation of LSPR of the printed GNS patterns even at low laser intensities. It was found that beside the direct role of the laser intensity, this effect depends appreciably on the printing parameters, such as drop density (δ, drops/mm2) and number of printed layers, and, critically, on the permeability of the coated paper substrates. These results will promote the development of GNS-based printed platforms for local photothermal therapy.

Gold Nanostar Synthesis and Functionalization with Organic Molecules

This chapter is devoted to the synthesis and functionalization of gold nanostars. The physical-chemical characterization and singular features of gold nanostars with respect to other types of gold nanoparticles are provided. Various methods of GNS synthesis as well as of the functionalization of GNS with PEG, organic dyes, and bioactive compounds are discussed.