Giacomo Dacarro

Antibacterial Activity of Glutathione-Coated Silver Nanoparticles against Gram Positive and Gram Negative Bacteria

In the present paper, we study the mechanism of antibacterial activity of glutathione (GSH) coated silver nanoparticles (Ag NPs) on model Gram negative and Gram positive bacterial strains. Interference in bacterial cell replication is observed for both cellular strains when exposed to GSH stabilized colloidal silver in solution, and microbicidal activity was studied when GSH coated Ag NPs are (i) dispersed in colloidal suspensions or (ii) grafted on thiol-functionalized glass surfaces. The obtained results confirm that the effect of dispersed GSH capped Ag NPs (GSH Ag NPs) on Escherichia coli is more intense because it can be associated with the penetration of the colloid into the cytoplasm, with the subsequent local interaction of silver with cell components causing damages to the cells. Conversely, for Staphylococcus aureus, since the thick peptidoglycan layer of the cell wall prevents the penetration of the NPs inside the cytoplasm, the antimicrobial effect is limited and seems related to the interaction with the bacterial surfaces. Experiments on GSH Ag NPs grafted on glass allowed us to elucidate more precisely the antibacterial mechanism, showing that the action is reduced because of GSH coating and the limitation of the translational freedom of NPs.

Self-assembled monolayers of silver nanoparticles firmly grafted on glass surfaces: Low Ag + release for an efficient antibacterial activity

A two-step, easy synthetic strategy in solution has been optimized to prepare authentic monolayers of silver nanoparticles (NP) on MPTS-modified glass surfaces, that were investigated by AFM imaging and by quantitative silver determination techniques. NP in the monolayers remain firmly grafted (i.e. not released) when the surfaces are exposed to air, water or in the physiological conditions mimicked by phosphate saline buffer, as UV-Vis spectroscopy and AFM studies demonstrate. About 15% silver release as Ag(+) ions has been found after 15days when the surfaces are exposed to water. The released silver cations are responsible of an efficient local microbicidal activity against Escherichia coli and Staphylococcus aureus bacterial strains.

Antibiofilm activity of a monolayer of silver nanoparticles anchored to an amino-silanized glass surface

Biofilm production is the crucial pathogenic mechanism of the implant-associated infection and a primary target for new anti-infective strategies. Silver nanoparticles (AgNPs) are attracting interest for their multifaceted potential biomedical applications. As endowed with highest surface/mass ratio and potent antibacterial activity, they can profitably be applied as monolayers at biomaterial surfaces. Desirably, in order to minimize the risks of toxic effects from freely circulating detached nanoparticles, AgNPs should firmly be anchored to the modified biomaterial surfaces. Here we focus on a newly designed glass surface modified with AgNPs and on its antibiofilm properties. Link of a self-assembled monolayer of AgNPs to glass was obtained through preliminary amino-silanization of the glass followed by immersion in an AgNPs colloidal suspension. Static contact angle measure, AFM, TEM, UV-Vis spectroscopy, ICP atomic emission spectroscopy were used for characterization. Antibiofilm activity against the biofilm-producer Staphylococcus epidermidis RP62A was assayed by both CFU method and CLSM. Performances of AgNPs-glasses were: i) excellent stability in aqueous medium; ii) prolonged release and high local concentration of Ag(+) without any detaching of AgNPs; iii) strong antibiofilm activity against S. epidermidis RP62A. This AgNPs surface-modification can be applied to a large variety of biomaterials by simply depositing glass-like SiO2 films on their surfaces.

Synthesis of branched Au nanoparticles with tunable near-infrared LSPR using a zwitterionic surfactant

Asymmetric branched gold nanoparticles are obtained using for the first time in the seed-growth approach a zwitterionic surfactant, laurylsulfobetaine, whose concentration in the growth solution allows to control both the length to base-width ratio of the branches and the LSPR position, that can be tuned in the 700-1100 nm near infrared range.

Self-assembled monolayers of gold nanostars: a convenient tool for near-IR photothermal biofilm eradication

Monolayers of gold nanostars (GNS) are grafted on mercaptopropyltrimethoxysilane-coated glass slides. In the formed monolayers the localized surface plasmon resonance of GNS can be tuned in the 700-1100 nm range. Upon laser excitation of the nearIR LSPR an efficient photothermal response is observed, inducing local hyperthermia and efficient killing of Staphylococcus aureus biofilms.

Spectroscopic evaluation of surface functionalization efficiency in the preparation of mercaptopropyltrimethoxysilane self-assembled monolayers on glass.

Four synthetic protocols have been examined for the preparation of a self-assembled monolayer of propanethiol functions on glass surfaces, through the wet reaction of mercaptopropyltrimethoxysilane (MPTS) on glass slides, in order to determine an optimal method to obtain dense -SH monolayers for further reaction with maleimide-bearing molecules. We have also demonstrated that the surface density of reactive -SH groups can be indagated by the coupling reaction with a maleimide-functionalized rhodamine dye and by the examination of the absorption spectra of the glass slides. The reaction of MPTS in dry toluene gives the most dense surface of reactive thiols, among the examined protocols. We have also evidenced that thermal curing of the MPTS functionalized slides induces thiol coupling and -S-S- formation, resulting in a dramatically lowered availability of -SH groups.

Strong modification of light emission from a dye monolayer via Bloch surface waves

We observe a strong modification of the emission properties of a rhodamine monolayer bonded to the surface of a 1D Si(1-x)N(x):H PhC. The photoluminescence signal observed at 670 nm is enhanced by more than 1 order of magnitude at the angle where the emitters are evanescently coupled to a Bloch surface wave supported by the structure. These results may be important for the design of efficient light-emitting devices and of optical sensors.

Monolayers of polyethilenimine on flat glass: a versatile platform for cations coordination and nanoparticles grafting in the preparation of antibacterial surfaces

A polyethylenimine (PEI) self-assembled monolayer (SAM) is prepared, capable of complexing silver and copper cations and of anchoring silver nanoparticles, exerting antibacterial activity against Escherichia coli and Staphylococcus aureus. Functionalized glassy surfaces have been fully characterized through spectroscopic techniques (UV-Vis spectroscopy, spectroscopic ellipsometry), atomic force microscopy imaging and quantitative Ag and Cu analysis (ICP optical emission spectroscopy).

Localized Surface Plasmon Resonance with Five-Branched Gold Nanostars in a Plastic Optical Fiber for Bio-Chemical Sensor Implementation

In this paper a refractive index sensor based on localized surface plasmon resonance (LSPR) in a Plastic Optical Fiber (POF), is presented and experimentally tested. LSPR is achieved exploiting five-branched gold nanostars (GNS) obtained using Triton X-100 in a seed-growth synthesis. They have the uncommon feature of three localized surface plasmon resonances. The strongest LSPRs fall in two ranges, one in the 600–900 nm range (LSPR 2) and the other one in the 1,100–1,600 nm range (LSPR 3), both sensible to refractive index changes. Anyway, due to the extremely strong attenuation (>102 dB/m) of the employed POF in the 1,100–1,600 nm range, only LSPR 2 will be exploited for refractive index change measurements, useful for bio-chemical sensing applications, as a proof of principle of the possibility of realizing a compact, low cost and easy-to-use GNS based device.

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.