Thermo-Plasmonic Killing of Escherichia coli TG1 Bacteria.

Plasmonic photo-thermal therapy (PPTT) is a minimally invasive, drug-free, therapy based on the properties of noble metal nanoparticles, able to convert a bio-transparent electromagnetic radiation into heat. PPTT has been used against cancer and other diseases. Herein, we demonstrate an antimicrobial methodology based on the properties of gold nanorods (GNRs). Under a resonant laser irradiation GNRs become highly efficient light to heat nano-converters extremely useful for PPTT applications. The concept here is to assess the antimicrobial effect of easy to synthesize, suitably purified, water-dispersible GNRs on Escherichia coli bacteria. A control on the GNRs concentration used for the process has been demonstrated critical in order to rule out cytotoxic effects on the cells, and still to be able to generate, under a near infrared illumination, an adequate amount of heat suited to increase the temperature up to ≈50 °C in about 5 min. Viability experiments evidenced that the proposed system accomplished a killing efficiency suitable to reducing the Escherichia coli population of about 2 log CFU (colony-forming unit).

Surface Engineering of Gold Nanorods for Cytochrome c Bioconjugation: An Effective Strategy To Preserve the Protein Structure.

The surface of gold nanorods (Au NRs) has been appropriately engineered to achieve a suitable interface for bioconjugation with horse heart cytochrome c (HCc). HCc, an extensively studied and well-characterized protein, represents an ideal model for nanoparticle (NP)-protein conjugation studies because of its small size, high stability, and commercial availability. Here, the native state of the protein has been demonstrated for the first time, by means of Raman spectroscopy, to be retained upon conjugation with the anisotropic Au nanostructures, thus validating the proposed protocol as specifically suited to mostly preserve the plasmonic properties of the NRs and to retain the structure of the protein. The successful creation of such bioconjugates with the retention of the protein structure and function along with the preservation of the NP properties represents a challenging but essential task, as it provides the only way to access functional hybrid systems with potential applications in biotechnology, medicine, and catalysis. In this perspective, the organic capping surrounding the Au NRs plays a key role, as it represents the functional interface for the conjugation step. Cetyltrimethylammonium bromide-coated Au NRs, prepared by using a seed-mediated synthetic route, have been wrapped with polyacrylic acid (PAA) by means of electrostatic interactions following a layer-by-layer approach. The resulting water-dispersible negatively charged AuNRs@PAA NPs have then been electrostatically bound to the positively charged HCc. The bioconjugation procedure has been thoroughly monitored by the combined analysis of UV-vis absorption, resonance Raman and Fourier transform infrared spectroscopies, transmission electron microscopy microscopy, and ζ-potential, which verified the successful conjugation of the protein to the nanorods.

Photo-Induced Heating in Plasmonic Nanoparticles Trapped in Thermo-Sensitive Liquid Crystals.

Thermo-sensitive liquid crystals may result, for some aspects, good host materials for plasmonic nanoparticles. In particular they are suitable to study and measure the temperature variations produced by photo-induced plasmonic joule effect in the metallic nanoparticles. Combining the properties of liquid crystals and metallic nanoparticles, allows to measure temperature variations in different ways by exploiting the optical properties of thermotropic liquid crystals: In a first attempt, by combining nematic liquid crystals and spherical metallic nanoparticles, we have predicted and measured temperature changes, under a suitable (resonant) optical illumination, by measuring the photo-thermal induced birefringence variation. In a different experiment, we have combined cholesteric liquid crystals and gold nanorods: Light-induced variations of structural colorations exhibited by cholesteric liquid crystals has been used as a new methodology to measure nanoscale heat variation with a very high sensitivity (0.03 K).

Gold-chlorophyll a-hybrid nanoparticles and chlorophyll a/cetyltrimethylammonium chloride self-assembled-suprastructures as novel carriers for chlorophyll a delivery in water medium: Photoactivity and photostability.

The stability of Chlorophyll a in water during prolonged exposure, at room temperature, to a neon lamp has been investigated by means of UV-vis and fluorescence spectroscopies. In addition, the Chlorophyll a (photo)stability evaluation in presence of suitable carriers has been performed in order to investigate its reactivity under the same conditions, for possible and future applications in Antimicrobial Photodynamic Therapy. Cetyltrimethylammonium chloride was chosen to solubilize Chlorophyll a in water. While, cetyltrimethylammonium chloride-capped gold nanoparticles offer a great opportunity because combine the Chlorophyll a action, used as a photosensitizer in Antimicrobial Photodynamic Therapy, with gold nanoparticles effect used in photothermal therapy. Indeed, the latter ones have exhibited an interesting rise of temperature if irradiated with visible light. Overall, both examined systems, cetyltrimethylammonium chloride/Chlorophyll a and gold nanoparticles/Chlorophyll a, were able to induce the Reactive Oxygen Species formation fundamental for a potential application in Antimicrobial Photodynamic Therapy.

Photo-thermal effects in gold nanoparticles dispersed in thermotropic nematic liquid crystals.

The last few years have seen a growing interest in the ability of metallic nanoparticles (MNPs) to control temperature at the nanoscale. Under a suitable optical radiation, MNPs feature an enhanced light absorption/scattering, thus turning into an ideal nano-source of heat, remotely controllable by means of light. In this framework, we report our recent efforts on modeling and characterizing the photo-thermal effects observed in gold nanoparticles (GNPs) dispersed in thermotropic Liquid Crystals (LCs). Photo-induced temperature variations in GNPs dispersed in Nematic LCs (NLCs) have been studied by implementing an ad hoc theoretical model based on the thermal heating equation applied to an anisotropic medium. Theoretical predictions have been verified by performing photo-heating experiments on a sample containing a small percentage of GNPs dispersed in NLCs. Both theory and experiments represent an important achievement in understanding the physics of heat transfer at the nanoscale, with applications ranging from photonics to nanomedicine.

Plasmonics Meets Biology through Optics.

Plasmonic metallic nanoparticles (NPs) represent a relevant class of nanomaterials, which is able to achieve light localization down to nanoscale by exploiting a phenomenon called Localized Plasmon Resonance. In the last few years, NPs have been proposed to trigger DNA release or enhance ablation of diseased tissues, while minimizing damage to healthy tissues. In view of the therapeutic relevance of such plasmonic NPs; a detailed characterization of the electrostatic interaction between positively charged gold nanorods (GNRs) and a negatively charged whole-genome DNA solution is reported. The preparation of the hybrid biosystem has been investigated as a function of DNA concentration by means of ζ-potential; hydrodynamic diameter and gel electrophoresis analysis. The results have pointed out the specific conditions to achieve the most promising GNRs/DNA complex and its photo-thermal properties have been investigated. The overall study allows to envisage the possibility to ingeniously combine plasmonic and biological materials and, thus, enable design and development of an original non invasive all-optical methodology for monitoring photo-induced temperature variation with high sensitivity.

SERS Properties of Gold Nanorods at Resonance with Molecular, Transverse, and Longitudinal Plasmon Excitations.

The amplification of Raman signals of the heteroaromatic cation 1-(N-methylpyrid-4-yl)-2-(N-methylpyrrol-2-yl)ethylene (PEP+)) bound to Au nanorods (NRs) was investigated at different excitation wavelengths to study the effect of the laser resonance with the absorption band of the PEP+ moiety and with the two plasmon oscillation modes of the NR. Two different PEP+ derivatives, differing in the length of the alkyl chain bearing the anchoring group, were used as target molecules. Raman spectra obtained exciting at 514 or at 785 nm (i.e., exciting the transverse or the longitudinal plasmon band) present a higher intensity than that at 488 nm suggesting a higher Raman amplification when the laser excitation wavelength is resonant with one of the two plasmon modes. Moreover, considering results of Discrete Dipole Approximation (DDA) calculations of the local field generated at the NR surface when either the transverse or the longitudinal plasmon modes are excited, we deduced that the resonance condition of the 514-nm laser excitation with the absorption band of the dye strongly contributes to the amplification of the Raman signal.

Electroactive layer-by-layer plasmonic architectures based on Au nanorods.

Nanostructured films based on Au nanorods (NRs) have been obtained by layer-by-layer (LbL) assembly driven by electrostatic interaction between metal nanoparticles and polyelectrolytes. Multilayer films have been fabricated by using LbL assembly of poly(sodium styrenesulfonate) (PSS) and positively charged Au NRs on a polyelectrolyte-modified substrate. The effect of fabrication parameters, including the nature of the substrate, the polyelectrolyte initial anchoring layer, and the number of layers has been investigated by means of UV-vis absorbance spectroscopy and atomic force microscopy (AFM). The results demonstrated the dependence of morphology and plasmonic features in the multilayered nanostructured architectures from the nature of the anchoring polyelectrolyte on the substrate, the number of layers, and the kind of NR mutual assembly. In addition, a study of the electrochemical activity at the solid/liquid interface has been carried out in order to assess charge transport through the NR multilayer by using two molecular probes in solution, namely, potassium ferricyanide, a common and well-established redox mediator with reversible behavior, and cytochrome C, a robust model redox protein. The presented systematic study of the immobilization of Au NRs opens the venue to several application areas, such as (bio)chemical sensing.

Photosystem II based multilayers obtained by electrostatic layer-by-layer assembly on quartz substrates.

Photosystem II (PSII) proteins from spinach leaves were immobilized onto quartz substrates according to the Layer-by-Layer (LbL) procedure, alternating protein to polyethylenimine (PEI) layers by exploiting electrostatic interactions. The effects of several factors, such as storage conditions, ageing of the PSII-modified substrates, as well as PSII concentration in buffer, on the quality of the prepared multilayers, were investigated by UV-vis Absorption Spectroscopy and Atomic Force Microscopy (AFM). A number of 13 layers was found to be optimal to guarantee intense PSII optical signals with homogeneous morphological distributions of proteins. The multilayers resulted stable if stored in contact with air at 4 °C, as observed by UV-vis Absorption spectra recorded after 48 h. The best results in terms of AFM images and electron transfer efficiency (measured by Hill Reaction assays) were gained by using 5.6 × 10(-7) M chlorophyll concentration, obtaining multilayers with the most ordered protein distributions and the highest electron transfer efficiency, i.e. 85% of an iso-absorbing PSII suspension. The results highlight the possibility to successfully immobilize PSII proteins, without considerable loss of bioactivity, thanks to the mild nature of the electrostatic LbL technique, opening up possibilities of applications in the bioelectrochemical energy conversion and biosensoristic fields.

Ion-directed assembly of gold nanorods: a strategy for mercury detection.

Water-soluble gold nanorods (Au NRs) have been functionalized with an N-alkylaminopyrazole ligand, 1-[2-(octylamino)ethyl]-3,5-diphenylpyrazole (PyL), that has been demonstrated able to coordinate heavy metal ions. The N-alkylaminopyrazole functionalized Au NRs have been characterized by electron microscopy and spectroscopic investigation and tested in optical detection experiments of different ions, namely, Zn(2+), Cd(2+), Hg(2+), Cu(2+), Pb(2+), and As(3+). In particular, the exposure of the functionalized NRs to increasing amounts of Hg(2+) ions has resulted in a gradual red-shift and broadening of the longitudinal plasmon band, up to 900 nm. Interestingly, a significantly different response has been recorded for the other tested ions. In fact, no significant shift in the longitudinal plasmon band has been observed for any of them, while a nearly linear reduction in the plasmon band intensity versus ion concentration in solution has been detected. The very high sensitivity for Hg(2+) with respect to other investigated ions, with a limit of detection of 3 ppt, demonstrates that the functionalization of Au NRs with PyL is a very effective method to be implemented in a reliable colorimetric sensing device, able to push further down the detection limit achieved by applying similar strategies to spherical Au NPs.

Biomaterials based on photosynthetic membranes as potential sensors for herbicides.

In this study, ultrathin film multilayers of Photosystem II-enriched photosynthetic membranes (BBY) were prepared and immobilized on quartz substrates by means of a Layer by Layer procedure exploiting electrostatic interactions with poly(ethylenimine) as polyelectrolyte. The biomaterials thus obtained were characterized by means of optical techniques and Atomic Force Microscopy, highlighting the fact that the Layer by Layer approach allowed the BBYs to be immobilized with satisfactory results. The activity of these hybrid materials was evaluated by means of optical assays based on the Hill Reaction, indicating that the biosamples, which preserved about 65% of their original activity even ten weeks after preparation, were both stable and active. Furthermore, an investigation of the biochips' sensitivity to the herbicide terbutryn, as a model analyte, gave interesting results: inhibition of photosynthetic activity was observed at terbutryn concentrations higher than 10(-7)M, thus evidencing the potential of such biomaterials in the environmental biosensor field.

The fate of silver ions in the photochemical synthesis of gold nanorods: an extended X-ray absorption fine structure analysis.

Water-soluble gold nanorods (Au NRs) were synthesized using a silver-ion mediated photochemical route under UV irradiation. Extended X-ray Absorption Fine Structure (EXAFS) measurements on the Ag K-edge were performed on samples obtained at different Ag/Au ratios and at increasing irradiation times in order to investigate the fate of silver ions during the growth of Au NRs. EXAFS measurements allowed to probe the chemical state and the local environment of silver in the final product. Experimental data suggest that Ag atoms are placed on top of the Au particles as metallic Ag(0), while no significant contribution to the EXAFS spectra comes from AgBr or other Ag(+) based species. The reported results strongly support the deposition of Ag(0) islands on the (110) surfaces of the Au particles, thus driving the anisotropic growth via the (111) surfaces.