Lucio Litti

Magneto‐Plasmonic Au‐Fe Alloy Nanoparticles Designed for Multimodal SERS‐MRI‐CT Imaging

Diagnostic approaches based on multimodal imaging are needed for accurate selection of the therapeutic regimens in several diseases, although the dose of administered contrast drugs must be reduced to minimize side effects. Therefore, large efforts are deployed in the development of multimodal contrast agents (MCAs) that permit the complementary visualization of the same diseased area with different sensitivity and different spatial resolution by applying multiple diagnostic techniques. Ideally, MCAs should also allow imaging of diseased tissues with high spatial resolution during surgical interventions. Here a new system based on multifunctional Au-Fe alloy nanoparticles designed to satisfy the main requirements of an ideal MCA is reported and their biocompatibility and imaging capability are described. The MCAs show easy and versatile surface conjugation with thiolated molecules, magnetic resonance imaging (MRI) and computed X-ray tomography (CT) signals for anatomical and physiological information (i.e., diagnostic and prognostic imaging), large Raman signals amplified by surface enhanced Raman scattering (SERS) for high sensitivity and high resolution intrasurgical imaging, biocompatibility, exploitability for in vivo use and capability of selective accumulation in tumors by enhanced permeability and retention effect. Taken together, these results show that Au-Fe nanoalloys are excellent candidates as multimodal MRI-CT-SERS imaging agents.

LDI-MS assisted by chemical-free gold nanoparticles: enhanced sensitivity and reduced background in the low-mass region.

Gold nanoparticles (AuNPs) assisted laser desorption ionization mass spectrometry (LDI-MS) emerged as an effective technique for the detection of analytes with high sensitivity. The surface chemistry and the size of AuNPs are the crucial parameters for lowering the detection limits and increasing the selectivity of LDI-MS. Here we show that chemical-free size selected AuNPs, obtained by laser ablation synthesis in solution (LASiS), have very low background in the low mass region (<500 Da), contrary to citrate stabilized AuNPs (citrate-AuNPs) and dihydroxyacetophenone (DHAP). This allowed better performances for the picomole detection of low mass analytes like arginine, fructose, atrazine, anthracene and paclitaxel. The results suggest that chemical-free LASiS-AuNPs can be an excellent matrix for nanoparticle-assisted LDI-MS.

Alternative SERRS probes for the immunochemical localization of ovalbumin in paintings: an advanced mapping detection approach.

In the field of analytical chemistry, many scientific efforts have been devoted to develop experimental procedures for the characterization of organic substances present in heterogeneous artwork samples, due to their challenging identification. In particular, performances of immunochemical techniques have been recently investigated, optimizing ad hoc systems for the identification of proteins. Among all the different immunochemical approaches, the use of metal nanoparticles - for surface enhanced Raman scattering (SERS) detection - remains one of the most powerful methods that has still not been explored enough for the analysis of artistic artefacts. For this reason, the present research work was aimed at proposing a new optimized and highly efficient indirect immunoassay for the detection of ovalbumin. In particular, the study proposed a new SERRS probe composed of gold nanoparticles (AuNPs) functionalised with Nile Blue A and produced with an excellent green and cheap alternative approach to the traditional chemical nanoparticles synthesis: the laser ablation synthesis in solution (LASiS). This procedure allows us to obtain stable nanoparticles which can be easily functionalized without any ligand exchange reaction or extensive purification procedures. Moreover, the present research work also focused on the development of a comprehensive analytical approach, based on the combination of potentialities of immunochemical methods and Raman analysis, for the simultaneous identification of the target protein and the different organic and inorganic substances present in the paint matrix. An advanced mapping detection system was proposed to achieve the exact spatial location of all the components through the creation of false colour chemical maps.

Use of nano gold obtained by laser ablation for SEIRA analyses of colorants

The analysis of dyes in cultural heritage samples is a well-known challenging task, due to their inherent high tinting strength and consequent low concentration in the carrying matrix a fact that severely limits the number of analytical techniques that can be efficiently and micro-destructively employed for their detection and unambiguous identification.In the present study, an advanced and alternative SEIRA based analytical protocol for the analysis of small quantities of synthetic colorants has been proposed.The method has been set up for the identification of Acid Orange 7 (AO7) using Au nanoparticles obtained by laser ablation in solution (LASiS). Analyses have been performed applying a drop containing a mixture between the colorant and the Au colloidal solution in its unaggregated state on a gold coated glass slide for RAS (Reflection Absorption Spectroscopy) analysis. The first results showed that, thanks to the enhancement produced by the nanoparticles, it is possible to analyze small amount of diluted solutions containing the colorant. Thus, the method has been successfully applied for the analysis of few pieces of dyed wool, after the development of a suitable micro extraction procedure.Graphical Abstractᅟ

A SERRS/MRI multimodal contrast agent based on naked Au nanoparticles functionalized with a Gd(III) loaded PEG polymer for tumor imaging and localized hyperthermia

Multimodal contrast agents offer new interesting diagnostic possibilities, summing the benefits of multiple imaging techniques. Magnetic resonance and optical imaging are complementary techniques. The first allows total body screening, even though it suffers from low spatial resolution and needs high loadings, whereas the second shows lower penetration, but bright signals, and a higher spatial resolution and needs lower loadings. We present a plasmonic nanosystem as a MRI (magnetic resonance imaging) and SERRS (surface enhanced resonance Raman scattering) multimodal contrast agent. Naked gold nanoparticles, obtained by laser ablation synthesis in solution, are organized as a highly efficient SERRS substrate with a naphthalocyanine reporter and functionalized with a MRI contrast agent with a newly synthesized 3DOTA-PEG polymer, with a high GdIII loading. As a proof of concept, in vivo and ex vivo MRI and SERRS experiments are also performed. The plasmonic property of the nanosystem is then exploited to show its usefulness for localized hyperthermia.

Safe core-satellite magneto-plasmonic nanostructures for efficient targeting and photothermal treatment of tumor cells

Magneto-plasmonic nanostructures functionalized with cell targeting units are of great interest for nanobiotechnology applications. Photothermal treatment of cells targeted with antibody functionalized nanostructures and followed by magnetic isolation, allows killing selected cells and hence is one of the applications of great interest. The magneto-plasmonic nanostructures reported herein were synthesized using naked gold and magnetite nanoparticles obtained through a green approach based on laser ablation of bulk materials in water. These particles do not need purifications steps for biocompatibility and are functionalized with a SERRS (surface enhanced resonance Raman scattering) active molecule for detection and with an antibody for targeting prostate tumor cells. Quantitative results for the cell targeting and selection efficiency show an overall accuracy of 94% at picomolar concentrations. The photothermal treatment efficiently kills targeted and magneto-selected cells producing a viability below 5% after 3 min of irradiation, compared with almost 100% viability of incubated and irradiated, but non targeted cells.

A new integrated TLC/MU-ATR/SERS advanced approach for the identification of trace amounts of dyes in mixtures

The present research is focused on the setting up of an advanced analytical system for the detection of synthetic dyes. The system is based on the combination of an innovative thin layer chromatography (TLC) plate coupled with enhanced infrared (MU-ATR, metal underlayer attenuated total reflection) and Surface Enhanced Raman (SERS) spectroscopy. In particular, a TLC plate made of silver iodide (AgI) applied onto a gold coated glass slide (AgI@Au) is proposed as an efficient stationary phase for the separation of dyes mixtures. The separated dyes are then identified by means of both enhanced FTIR and SERS, performed directly on the same eluted spots. The use of a mid-IR transparent inorganic salt as stationary phase coupled with the underneath gold layer avoids spectral interferences, enhancing the signal obtained from ATR analyses. At the same time, SERS spectra can be recorded as the TLC plate may act as a SERS active substrate due to the photoreduction of AgI to metallic Ag caused by the exposure to the laser during the Raman analysis. Different mixtures of synthetic dyes of known composition, widely used in dyeing processes, have been tested and the method resulted to be effective in identifying trace amounts in the order of tens nanograms. Moreover, the method has been further evaluated on a real case study represented by dyes extracted from dyed wool.

Enhanced EGFR Targeting Activity of Plasmonic Nanostructures with Engineered GE11 Peptide

Plasmonic nanostructures show important properties for biotechnological applications, but they have to be guided on the target for exploiting their potentialities. Antibodies are the natural molecules for targeting. However, their possible adverse immunogenic activity and their cost have suggested finding other valid substitutes. Small molecules like peptides can be an alternative source of targeting agents, even if, as single molecules, their binding affinity is usually not very good. GE11 is a small dodecapeptide with specific binding to the epidermal growth factor receptor (EGFR) and low immunogenicity. The present work shows that thousands of polyethylene glycol (PEG) chains modified with lysines and functionalized with GE11 on clusters of naked gold nanoparticles, obtained by laser ablation in water, achieves a better targeting activity than that recorded with nanoparticles decorated with the specific anti-EGFR antibody Cetuximab (C225). The insertion of the cationic spacer between the polymeric part of the ligand and the targeting peptide allows for a proper presentation of GE11 on the surface of the nanosystems. Surface enhanced resonance Raman scattering signals of the plasmonic gold nanoparticles are used for quantifying the targeting activity. Molecular dynamic calculations suggest that subtle differences in the exposition of the peptide on the PEG sea are important for the targeting activity.

A surface enhanced Raman scattering based colloid nanosensor for developing therapeutic drug monitoring

Competitive reactions, on the surface of plasmonic nanostructures, allow exploiting SERS signals for quantitative Therapeutic Drug Monitoring. As an example, the concentration of Erlotinib, an anti-EGFR small molecule, used for the treatment of non-small cell lung and pancreatic cancer, is determined. The numerous side effects and the variability of patient responses make Erlotinib a good candidate for monitoring. The new SERS based sensor can estimate Erlotinib down to nanomolar concentration and is based on the chemical reaction of the drug and of a competitor SERS reporter on the surface of gold nanostructures. Colloid solutions of naked gold nanoparticles obtained by laser ablation in solution were used for obtaining nanostructures with very efficient hot spots for SERS and with a clean surface for chemistry. Detection of the drug in the nanomolar concentration range is shown to be possible also in spiked plasma samples.

Single File Flow of Biomimetic Beads for Continuous SERS Recording in a Microfluidic Device

A major challenge in cancer treatment is the quantification of biomarkers associated with a specific cancer type. Important biomarkers are the circulating tumor cells (CTCs) detached from the main cancer and circulating in the blood. CTCs are very rare and their identification is still an issue. Although CTCs quantification can be estimated by using fluorescent markers, all the fluorescence techniques are strongly limited by the number of emissions (therefore markers) that can be discriminated with one exciting line, by their bleaching characteristics, and by the intrinsic autofluorescence of biological samples. An emerging technique that can overcome these limitations is Surface Enhanced Raman Scattering (SERS). Signals of vibrational origin with intensity similar to those of fluorescence, but narrower bandwidths, can be easily discriminated even by exciting with a single laser line. We recently showed the benefit of this method with cells fixed on a surface. However, this approach is too demanding to be applied in clinical routine. To effectively increase the throughput of the SERS analysis, microfluidics represents a promising tool. We report two different hydrodynamic strategies, based on device geometry and liquids viscosity, to successfully combine a microfluidic design with SERS.