Photomobile Polymer-Piezoelectric Composite for Enhanced Actuation and Energy Generation.

In this study, we present an innovative approach to increase the quantum yield and wavelength sensitivity of photomobile polymer (PMP) films based on azobenzene by doping the polymer matrix with noble metal nanoparticles. These doped PMP films showed faster and more significant bending under both UV as well as visible and near-infrared light regardless of whether it was coherent, incoherent, polarized, or unpolarized irradiation, expanding the potential of PMP-based actuators. To illustrate their practical implications, we created a proof-of-concept model of power generation by coupling it to flexible piezoelectric materials under simulated sunlight. This model has been tested under real operating conditions, thus demonstrating the possibility of generating electricity with variable light exposure. Additionally, our synthetic protocol is solvent-free, which is another benefit of environmental relevance. Our research lays the groundwork for the development of sunlight-sensitive devices, such as photomechanical actuators and advanced photovoltaic modules, which may break ground in the thriving field of smart materials. We are confident that the presented findings will contribute to the ongoing discourse in the field and inspire additional advances in renewable energy applications.

Cyanine-Doped Nanofiber Mats for Laser Tissue Bonding.

In spite of an extensive body of academic initiatives and innovative products, the toolkit of wound dressing has always revolved around a few common concepts such as adhesive patches and stitches and their variants. Our work aims at an alternative solution for an immediate restitutio ad integrum of the mechanical functionality in cutaneous repairs. We describe the fabrication and the application of electrospun mats of bioactive nanofibers all made of biocompatible components such as a natural polysaccharide and a cyanine dye for use as laser-activatable plasters, resembling the ultrastructure of human dermis. In particular, we investigate their morphological features and mechanical moduli under conditions of physiological relevance, and we test their use to bind a frequent benchmark of connective tissue as rabbit tendon and a significant case of clinical relevance as human dermis. Altogether, our results point to the feasibility of a new material for wound dressing combining translational potential, strength close to human dermis, extensibility exceeding 15% and state-of-art adhesive properties.

Paper-based genetic assays with bioconjugated gold nanorods and an automated readout pipeline.

Paper-based biosensors featuring immunoconjugated gold nanoparticles have gained extraordinary momentum in recent times as the platform of choice in key cases of field applications, including the so-called rapid antigen tests for SARS-CoV-2. Here, we propose a revision of this format, one that may leverage on the most recent advances in materials science and data processing. In particular, we target an amplifiable DNA rather than a protein analyte, and we replace gold nanospheres with anisotropic nanorods, which are intrinsically brighter by a factor of ~ 10, and multiplexable. By comparison with a gold-standard method for dot-blot readout with digoxigenin, we show that gold nanorods entail much faster and easier processing, at the cost of a higher limit of detection (from below 1 to 10 ppm in the case of plasmid DNA containing a target transgene, in our current setup). In addition, we test a complete workflow to acquire and process photographs of dot-blot membranes with custom-made hardware and regression tools, as a strategy to gain more analytical sensitivity and potential for quantification. A leave-one-out approach for training and validation with as few as 36 sample instances already improves the limit of detection reached by the naked eye by a factor around 2. Taken together, we conjecture that the synergistic combination of new materials and innovative tools for data processing may bring the analytical sensitivity of paper-based biosensors to approach the level of lab-grade molecular tests.

Microbubble resonators for scattering-free absorption spectroscopy of nanoparticles.

We present a proof-of-concept experiment where the absorbance spectra of suspensions of plasmonic nanoparticles are accurately reconstructed through the photothermal conversion that they mediate in a microbubble resonator. This thermal detection produces spectra that are insensitive towards light scattering in the sample, as proved experimentally by comparing the spectra of acqueos gold nanorods suspensions in the presence or absence of milk powder. In addition, the microbubble system allows for the interrogation of small samples (below 40 nl) while using a low-intensity beam (around 20 µW) for their excitation. In perspective, this system could be implemented for the characterization of turbid biological fluids through their optical absorption, especially when considering that the microbubble resonator naturally interfaces to a microfluidic circuit and may easily fit within portable or on-chip devices.

Enhanced Antitumoral Activity and Photoacoustic Imaging Properties of AuNP-Enriched Endothelial Colony Forming Cells on Melanoma.

Near infrared (NIR)-resonant gold nanoparticles (AuNPs) hold great promise in cancer diagnostics and treatment. However, translating the theranostic potential of AuNPs into clinical applications still remains a challenge due to the difficulty to improve the efficiency and specificity of tumor delivery in vivo as well as the clearance from liver and spleen to avoid off target toxicity. In this study, endothelial colony forming cells (ECFCs) are exploited as vehicles to deliver AuNPs to tumors. It is first demonstrated that ECFCs display a great capability to intake AuNPs without losing viability, and exert antitumor activity per se. Using a human melanoma xenograft mouse model, it is next demonstrated that AuNP-loaded ECFCs retain their capacity to migrate to tumor sites in vivo 1 day after injection and stay in the tumor mass for more than 1 week. In addition, it is demonstrated that ECFC-loaded AuNPs are efficiently cleared by the liver over time and do not elicit any sign of damage to healthy tissue.

Photostability of Contrast Agents for Photoacoustics: The Case of Gold Nanorods.

Plasmonic particles as gold nanorods have emerged as powerful contrast agents for critical applications as the photoacoustic imaging and photothermal ablation of cancer. However, their unique efficiency of photothermal conversion may turn into a practical disadvantage, and expose them to the risk of overheating and irreversible photodamage. Here, we outline the main ideas behind the technology of photoacoustic imaging and the use of relevant contrast agents, with a main focus on gold nanorods. We delve into the processes of premelting and reshaping of gold nanorods under illumination with optical pulses of a typical duration in the order of few ns, and we present different approaches to mitigate this issue. We undertake a retrospective classification of such approaches according to their underlying, often implicit, principles as: constraining the initial shape; or speeding up their thermal coupling to the environment by lowering their interfacial thermal resistance; or redistributing the input energy among more particles. We discuss advantages, disadvantages and contexts of practical interest where one solution may be more appropriate than the other.

A bionic shuttle carrying multi-modular particles and holding tumor-tropic features.

The systemic delivery of composite nanoparticles remains an outstanding challenge in cancer nanomedicine, and the principal reason is a complex interplay of biological barriers. In this regard, adaptive cell transfer may represent an alternative solution to circumvent these barriers down to the tumor microenvironment. Here, tumor-tropic macrophages are proposed as a tool to draw and vehiculate modular nanoparticles integrating magnetic and plasmonic components. The end result is a bionic shuttle that exhibits a plasmonic band within the so-called therapeutic window arising from as much as 40 pg Au per cell, magnetization in the order of 150 pemu per cell, and more than 90% of the pristine viability and chemotactic activity of its biological component, until at least two days of preparation. Its synergistic combination of plasmonic, magnetic and tumor-tropic functions is assessed in vitro for applications as magnetic guidance or sorting, with a propulsion around 4 µm s-1 for a magnetic gradient of 0.8 T m-1, the optical hyperthermia of cancer, with stability of photothermal conversion to temperatures exceeding 50∘C, and the photoacoustic imaging of cancer under realistic conditions. These results collectively suggest that a bionic design may be a promising roadmap to reconcile the efforts for multifunctionality and targeted delivery, which are both key goals in nanomedicine.

Impact of Kapitza resistance on the stability and efficiency of photoacoustic conversion from gold nanorods.

Plasmonic particles have been proposed for a broad variety of optical and hybrid applications, including the photothermal ablation and photoacoustic imaging of cancer, or their integration in photonic sensors. Here, we address the effect of thermal resistance at the gold-water interface, or Kapitza resistance, on the performance of photoacoustic conversion of gold nanorods. Our findings point to possible strategies for the optimization of plasmonic particles as contrast agents for imaging, or even as transducers for biosensing. We perform numerical simulations that project a simultaneous increase of efficiency and stability of photoacoustic conversion with a decrease of Kapitza resistance. We suggest an effective approach to modulate Kapitza resistance by including underresolved features as roughness or the presence of adsorbates. Inspired by this idea, we synthesize a rough variant of gold nanorods by the deposition and galvanic replacement of a silver shell, where roughness provides higher photoacoustic signals by about 70% and damage thresholds by 120%. In addition, we coat our particles with a protein corona and find a decrease of photoacoustic signals with shell thickness, which may inspire new solutions for biosensors based on a mechanism of photoacoustic transduction. Both our findings are consistent with an effective modulation of Kapitza resistance, which decreases upon roughening, due to an underlying increase of specific surface area, and increases upon coating with a protein shell that may act as a thermal insulation. We discuss possible directions to gain more advantage of our concept for topical applications at the crossroads of plasmonics, biomedical optics and biosensing.

Optically activated and interrogated plasmonic hydrogels for applications in wound healing.

We disclose the use of hybrid materials featuring Au/Ag core/shell nanorods in porous chitosan/polyvinyl alcohol scaffolds for applications in tissue engineering and wound healing. The combination of Au and Ag in a single construct provides synergistic opportunities for optical activation of functions as near infrared laser tissue bonding, and remote interrogation to return parameters of prognostic relevance in wound healing monitoring. In particular, the bimetallic component ensures optical tunability, enhanced shelf life and photothermal stability, serves as a reservoir of germicidal silver cations, and changes in near-infrared and visible color according to the environmental level of oxidative stress. At the same time, the polymeric blend is ideal to bind connective tissue upon photothermal activation, and to support fabrication processes that provide high porosity, such as electrospinning, thus putting all the premises for cellular repopulation and antimicrobial protection.

Microbubble Resonators for All-Optical Photoacoustics of Flowing Contrast Agents.

In this paper, we implement a Whispering Gallery mode microbubble resonator (MBR) as an optical transducer to detect the photoacoustic (PA) signal generated by plasmonic nanoparticles. We simulate a flow cytometry experiment by letting the nanoparticles run through the MBR during measurements and we estimate PA intensity by a Fourier analysis of the read-out signal. This method exploits the peaks associated with the MBR mechanical eigenmodes, allowing the PA response of the nanoparticles to be decoupled from the noise associated with the particle flow whilst also increasing the signal-to-noise ratio. The photostability curve of a known contrast agent is correctly reconstructed, validating the proposed analysis and proving quantitative PA detection. The experiment was run to demonstrate the feasible implementation of the MBR system in a flow cytometry application (e.g., the detection of venous thrombi or circulating tumor cells), particularly regarding wearable appliances. Indeed, these devices could also benefit from other MBR features, such as the extreme compactness, the direct implementation in a microfluidic circuit, and the absence of impedance-matching material.

Fiber-cap biosensors for SERS analysis of liquid samples.

Optical detection techniques based on surface enhanced Raman spectroscopy (SERS) are a powerful tool for biosensing applications. Meanwhile, due to technological advances, different approaches have been investigated to integrate SERS substrates on the tip of optical fibres for molecular probing in liquids. To further demonstrate the perspectives offered by SERS-on-fiber technology for diagnostic purposes, in this study, novel cap-shaped SERS sensors for reversible coupling with customized multimodal probes were prototyped via low-cost polymer casting of polydimethylsiloxane (PDMS) and further assembly of gold nanoparticles (Au NPs) of varied sizes and shapes. To demonstrate the feasibility of liquid sensing with cap sensors using backside illumination and detection, the spectra of rhodamine were acquired by coupling the caps with the fiber. As expected by UV-vis, the highest SERS efficiency was observed for NP-decorated substrates with plasmonic properties in resonance with the irradiation wavelength. Then, SERS biosensors for the specific detection of amyloid-ß (Aß) neurotoxic biomarkers were realized by covalent grafting of Aß antibodies. As attested by fluorescence images and SERS measurements, the biosensors successfully exhibited enhanced Aß affinity compared to the bare sensors without ligands. Finally, these versatile (bio)sensors are a powerful tool to transform any milli-sized fibers into functional (bio)sensing platforms with plasmonic and biochemical properties tailored for specific applications.

Hybrid organosilicon/polyol phantom for photoacoustic imaging.

The rapid development of hardware and software for photoacoustic technologies is urging the establishment of dedicated tools for standardization and performance assessment. In particular, the fabrication of anatomical phantoms for photoacoustic imaging remains an open question, as current solutions have not yet gained unanimous support. Here, we propose that a hybrid material made of a water-in-oil emulsion of glycerol and polydimethylsiloxane may represent a versatile platform to host a broad taxonomy of hydrophobic and hydrophilic dyes and recapitulate the optical and acoustic features of bio tissue. For a full optical parameterization, we refer to Wróbel, et al. [ Biomed. Opt. Express7, 2088 (2016)], where this material was first presented for optical imaging. Instead, here, we complete the picture and find that its speed of sound and acoustic attenuation resemble those of pure polydimethylsiloxane, i.e. respectively 1150 ± 30 m/s and 3.5 ± 0.4 dB/(MHz·cm). We demonstrate its use under a commercial B-mode scanner and a home-made A-mode stage for photoacoustic analysis to retrieve the ground-truth encoded in a multilayer architecture containing indocyanine green, plasmonic particles and red blood cells. Finally, we verify the stability of its acoustic, optical and geometric features over a time span of three months.

1064-nm-resonant gold nanorods for photoacoustic theranostics within permissible exposure limits.

Therapeutic and diagnostic methods based on photomechanical effects are attracting much current attention in contexts as oncology, cardiology and vascular surgery, for such applications as photoacoustic imaging or microsurgery. Their underlying mechanism is the generation of ultrasound or cavitation from the interaction of short optical pulses with endogenous dyes or targeted contrast agents. Among the latter, gold nanorods are outstanding candidates, but their use has mainly been reported for photoacoustic imaging and photothermal treatments. Conversely, much less is still known about their value as a precision tool for photomechanical manipulations, such as to impart local damage with high spatial resolution through the expansion and collapse of microbubbles. Here, we address the feasibility of gold nanorods exhibiting a distribution of surface plasmon resonances between about 900 to above 1100 nm as a contrast agent for photoacoustic theranostics. After testing their cytotoxicity and cellular uptake, we discuss their photostability and use to mediate cavitation and the photomechanical destruction of targeted cells. We find that the choice of a plasmonic band peaking around 1064 nm is key to enhance the translational potential of this approach. With respect to the standard alternative of 800 nm, at 1064 nm, relevant regulations on optical exposure are less restrictive and the photonic technology is more mature.

Polylysine as a functional biopolymer to couple gold nanorods to tumor-tropic cells.

BACKGROUND: The delivery of plasmonic particles, such as gold nanorods, to the tumor microenvironment has attracted much interest in biomedical optics for topical applications as the photoacoustic imaging and photothermal ablation of cancer. However, the systemic injection of free particles still crashes into a complexity of biological barriers, such as the reticuloendothelial system, that prevent their efficient biodistribution. In this context, the notion to exploit the inherent features of tumor-tropic cells for the creation of a Trojan horse is emerging as a plausible alternative. RESULTS: We report on a convenient approach to load cationic gold nanorods into murine macrophages that exhibit chemotactic sensitivity to track gradients of inflammatory stimuli. In particular, we compare a new model of poly-L-lysine-coated particles against two alternatives of cationic moieties that we have presented elsewhere, i.e. a small quaternary ammonium compound and an arginine-rich cell-penetrating peptide. Murine macrophages that are exposed to poly-L-lysine-coated gold nanorods at a dosage of 400 µM Au for 24 h undertake efficient uptake, i.e. around 3 pg Au per cell, retain the majority of their cargo until 24 h post-treatment and maintain around 90% of their pristine viability, chemotactic and pro-inflammatory functions. CONCLUSIONS: With respect to previous models of cationic coatings, poly-L-lysine is a competitive solution for the preparation of biological vehicles of gold nanorods, especially for applications that may require longer life span of the Trojan horse, say in the order of 24 h. This biopolymer combines the cost-effectiveness of small molecules and biocompatibility and efficiency of natural peptides and thus holds potential for translational developments.

A multifunctional organosilica cross-linker for the bio-conjugation of gold nanorods.

We report on the use of organosilica shells to couple gold nanorods to functional peptides and modulate their physiochemical and biological profiles. In particular, we focus on the case of cell penetrating peptides, which are used to load tumor-tropic macrophages and implement an innovative drug delivery system for photothermal and photoacoustic applications. The presence of organosilica exerts subtle effects on multiple parameters of the particles, including their size, shape, electrokinetic potential, photostability, kinetics of endocytic uptake and cytotoxicity, which are investigated by the interplay of colorimetric methods and digital holographic microscopy. As a rule of thumb, as the thickness of organosilica increases from none to ∼30nm, we find an improvement of the photophysical performances at the expense of a deterioration of the biological parameters. Therefore, detailed engineering of the particles for a certain application will require a careful trade-off between photophysical and biological specifications.

Statistical detection of nanoparticles in cells by darkfield microscopy.

In the fields of nanomedicine, biophotonics and radiation therapy, nanoparticle (NP) detection in cell models often represents a fundamental step for many in vivo studies. One common question is whether NPs have or have not interacted with cells. In this context, we propose an imaging based technique to detect the presence of NPs in eukaryotic cells. Darkfield images of cell cultures at low magnification (10×) are acquired in different spectral ranges and recombined so as to enhance the contrast due to the presence of NPs. Image analysis is applied to extract cell-based parameters (i.e. mean intensity), which are further analyzed by statistical tests (Student's t-test, permutation test) in order to obtain a robust detection method. By means of a statistical sample size analysis, the sensitivity of the whole methodology is quantified in terms of the minimum cell number that is needed to identify the presence of NPs. The method is presented in the case of HeLa cells incubated with gold nanorods labeled with anti-CA125 antibodies, which exploits the overexpression of CA125 in ovarian cancers. Control cases are considered as well, including PEG-coated NPs and HeLa cells without NPs.

Preparation and Photoacoustic Analysis of Cellular Vehicles Containing Gold Nanorods.

Gold nanorods are attractive for a range of biomedical applications, such as the photothermal ablation and the photoacoustic imaging of cancer, thanks to their intense optical absorbance in the near-infrared window, low cytotoxicity and potential to home into tumors. However, their delivery to tumors still remains an issue. An innovative approach consists of the exploitation of the tropism of tumor-associated macrophages that may be loaded with gold nanorods in vitro. Here, we describe the preparation and the photoacoustic inspection of cellular vehicles containing gold nanorods. PEGylated gold nanorods are modified with quaternary ammonium compounds, in order to achieve a cationic profile. On contact with murine macrophages in ordinary Petri dishes, these particles are found to undergo massive uptake into endocytic vesicles. Then these cells are embedded in biopolymeric hydrogels, which are used to verify that the stability of photoacoustic conversion of the particles is retained in their inclusion into cellular vehicles. We are confident that these results may provide new inspiration for the development of novel strategies to deliver plasmonic particles to tumors.

Tuning the interactions of PEG-coated gold nanorods with BSA and model proteins through insertion of amino or carboxylate groups.

Gold nanorods (GNRs) are important platforms for biosensing and drug delivery. As for most nanomaterials, appropriate coatings such as polyethylene glycol (PEG) are needed to stabilize GNRs within biological fluids. We show here that the interactions of GNRs with proteins can be finely modulated through surface modification using PEG-containing chains bearing charged headgroups. Interestingly, introduction of amino or carboxylate groups produces relevant and differential changes in GNR interactions with three representative proteins: lysozyme, cytochrome c, and bovine serum albumin. These effects were explored through the direct monitoring of plasmonic bands of the GNRs and are supported by independent dynamic light scattering (DLS) and circular dichroism (CD) determinations. Notably, GNR-protein interactions observed for these charged GNRs can be almost completely reversed by salt addition. These observations demonstrate the importance of electrostatic effects in governing GNR-protein interactions, and provide a basis for new sensing and delivery platforms.

Concave gold nanocube assemblies as nanotraps for surface-enhanced Raman scattering-based detection of proteins.

SERS detection of proteins is typically performed by using labeling agents with stable and high Raman scattering cross sections. This is a valuable approach for trace detection and quantification of a target protein but is unsuitable for inspecting its inherent structural and functional properties. On the other hand, direct SERS of proteins has been mainly devoted to the study of short peptides and aminoacid sequences or of prosthetic groups with intense Raman signals, which is of scarce interest for a thorough characterization of most proteins. Here we try to overcome these limitations by setting-up an effective platform for the structural SERS analysis of proteins. The platform consists of an extended bidimensional array of gold concave nanocubes (CNCs) supported on a PDMS film. CNCs are closely-packed through face-face and face-corner interactions generating a monolayered arrangement featuring well distributed nanoholes. Here the protein homogeneously experiences an E-field enhancement outward from the metal surfaces surrounding it, which causes a large number of vibrations to be contemporarily amplified. The proposed platform provides stable and detailed SERS spectra and confers rapidity and reproducibility to the analysis.

Detection of biomarkers for inflammatory diseases by an electrochemical immunoassay: the case of neopterin.

An electrochemical immunoassay for neopterin was developed using recently produced specific antibodies immobilized to protein A-coated magnetic beads in combination with differential pulse voltammetry and screen-printed array of electrodes. Neopterin-alkaline phosphatase conjugate was used as label in a competitive assay format. Multiplexed analysis of neopterin was demonstrated by replacing the traditional ELISA with electrochemical detection and the traditional plastic wells with screen-printed array of electrodes. The optimized electrochemical method, based on polyclonal antibodies, reached a limit of detection of 0.008 ng/mL with an average RSD %=10. Serum samples collected from patients with sepsis, healthy volunteers and other patients without a confirmed clinical diagnosis were also analyzed. The obtained results, compared with those of a commercial ELISA kit, had a significant correlation, showing the possibility to distinguish among the serum samples from ill or healthy subjects.