Dopamine-Coated Carbon Nanodots: A Supramolecular Approach to Polydopamine Composite.

The development of biocompatible composites constituted by polydopamine and fluorescent carbon dots represents a promising way of exploiting the extraordinary adhesive properties of polydopamine for multi-purpose technologies. Here, a supramolecular complex is realized by the assembly of dopamine on the carbon dots surface, and the optical and structural properties are investigated by means of different spectroscopic techniques, from time-resolved fluorescence to Raman and NMR spectroscopies. The results suggest that the catechol unit of dopamine plays the main role in the formation of the supramolecular complex, in which carbon nanodot fluorescence emission is quenched by a photoinduced electron transfer process. The interaction with the nanodots' basic surface sites promotes the oxidation of dopamine and drives to its oligomerization/polymerization on the nanodot surface.

KLVFF oligopeptide-decorated amphiphilic cyclodextrin nanomagnets for selective amyloid beta recognition and fishing.

Recognition and capture of amyloid beta (Aß) is a challenging task for the early diagnosis of neurodegenerative disorders, such as Alzheimer's disease. Here, we report a novel KLVFF-modified nanomagnet based on magnetic nanoparticles (MNP) covered with a non-ionic amphiphilic ß-cyclodextrin (SC16OH) and decorated with KLVFF oligopeptide for the self-recognition of the homologous amino-acids sequence of Aß to collect Aß (1-42) peptide from aqueous samples. MNP@SC16OH and MNP@SC16OH/Ada-Pep nanoassemblies were fully characterized by complementary techniques both as solid powders and in aqueous dispersions. Single domain MNP@SC16OH/Ada-Pep nanomagnets of 20-40 nm were observed by TEM analysis. DLS and ζ-potential measurements revealed that MNP@SC16OH nanoassemblies owned in aqueous dispersion a hydrodynamic radius of about 150 nm, which was unaffected by Ada-Pep decoration, while the negative ζ-potential of MNP@SC16OH (-40 mV) became less negative (-30 mV) in MNP@SC16OH/Ada-Pep, confirming the exposition of positively charged KLVFF on nanomagnets surface. The ability of MNP@SC16OH/Ada-Pep to recruit Aß (1-42) in aqueous solution was evaluated by MALDI-TOF and compared with the ineffectiveness of undecorated MNP@SC16OH and VFLKF scrambled peptide-decorated nanoassemblies (MNP@SC16OH/Ada-scPep), pointing out the selectivity of KLVFF-decorated nanohybrid towards Aß (1-42). Finally, the property of nanomagnets to extract Aß in conditioned medium of cells over-producing Aß peptides was investigated as proof of concept of effectiveness of these nanomaterials as potential diagnostic tools.

Water-Soluble Non-Ionic PEGylated Porphyrins: A Versatile Category of Dyes for Basic Science and Applications.

This review arises from the need to rationalize the huge amount of information on the structural and spectroscopic properties of a peculiar class of porphyrin derivatives-the non-ionic PEGylated porphyrins-collected during almost two decades of research. The lack of charged groups in the molecular architecture of these porphyrin derivatives is the leitmotif of the work and plays an outstanding role in highlighting those interactions between porphyrins, or between porphyrins and target molecules (e.g., hydrophobic-, hydrogen bond related-, and coordination-interactions, to name just a few) that are often masked by stronger electrostatic contributions. In addition, it is exactly these weaker interactions between porphyrins that make the aggregated forms more prone to couple efficiently with external perturbative fields like weak hydrodynamic vortexes or temperature gradients. In the absence of charge, solubility in water is very often achieved by covalent functionalization of the porphyrin ring with polyethylene glycol chains. Various modifications, including of chain length or the number of chains, the presence of a metal atom in the porphyrin core, or having two or more porphyrin rings in the molecular architecture, result in a wide range of properties. These encompass self-assembly with different aggregate morphology, molecular recognition of biomolecules, and different photophysical responses, which can be translated into numerous promising applications in the sensing and biomedical field, based on turn-on/turn-off fluorescence and on photogeneration of radical species.

Porphyrin-Based Supramolecular Flags in the Thermal Gradients' Wind: What Breaks the Symmetry, How and Why.

The Spontaneous Symmetry Breaking (SSB) phenomenon is a natural event in which a system changes its symmetric state, apparently reasonless, in an asymmetrical one. Nevertheless, this occurrence could be hiding unknown inductive forces. An intriguing investigation pathway uses supramolecular aggregates of suitable achiral porphyrins, useful to mimic the natural light-harvesting systems (as chlorophyll). Using as SSB probe supramolecular aggregates of 5,10,15,20-tetrakis[p(ω-methoxypolyethyleneoxy)phenyl]porphyrin (StarP), a non-ionic achiral PEGylated porphyrin, we explore here its interaction with weak asymmetric thermal gradients fields. The cross-correlation of the experimental data (circular dichroism, confocal microscopy, atomic force microscopy, and cryo-transmission electron microscopy) revealed that the used building blocks aggregate spontaneously, organizing in flag-like structures whose thermally-induced circular dichroism depends on their features. Finally, thermal gradient-induced enantioselectivity of the supramolecular flag-like aggregates has been shown and linked to their size-dependence mesoscopic deformation, which could be visualized as waving flags in the wind.

En Route to a Chiral Melanin: The Dynamic "From-Imprinted-to-Template" Supramolecular Role of Porphyrin Hetero-Aggregates During the Oxidative Polymerization of L-DOPA.

Chiral porphyrin hetero-aggregates, produced from meso-tetrakis(4-N-methylpyridyl) porphyrin H2T4 and copper(II) meso-tetrakis(4-sulfonatophenyl)porphyrin CuTPPS by an imprinting effect in the presence of L-3,4-dihydroxyphenylalanine (L-DOPA), are shown herein to serve as templates for the generation of chiral structures during the oxidative conversion of the amino acid to melanin. This remarkable phenomenon is suggested to involve the initial role of L-DOPA and related chiral intermediates like dopachrome as templates for the production of chiral porphyrin aggregates. When the entire chiral pool from DOPA is lost, chiral porphyrin hetero-aggregate would elicit axially chiral oligomer formation from 5,6-dihydroxyindole intermediates in the later stages of melanin synthesis. These results, if corroborated by further studies, may open unprecedented perspectives for efficient strategies of asymmetric melanin synthesis with potential biological and technological applications.

Interpenetrating Polymer Network Microgels in Water: Effect of Composition on the Structural Properties and Electrosteric Interactions.

Microgels of cross-linked interpenetrating polymer networks (IPNs) are very versatile systems combining the properties of colloids and polymers. Herein we study IPN microgels composed of poly(N-isopropylacrylamide) and poly(acrylic acid) to understand how weight composition and reactant concentrations affect their structural, conformational and electrosteric properties in water. The results show that it is possible to drive the formation of microgels with the desired properties by adjusting IPN composition and preparation method during the synthesis. During synthesis, the polymerization of acrylic acid triggers the merging among IPNs via covalent linking, giving rise to microgels with larger mass and size, the effect being larger for higher concentration of the reactants. In addition, a close relation between the microgel internal conformation and the colloidal stability is observed, due to the presence of screened groups inside the microgel.

Design principles of chiral carbon nanodots help convey chirality from molecular to nanoscale level.

The chirality of (nano)structures is paramount in many phenomena, including biological processes, self-assembly, enantioselective reactions, and light or electron spin polarization. In the quest for new chiral materials, metallo-organic hybrids have been attractive candidates for exploiting the aforementioned scientific fields. Here, we show that chiral carbon nanoparticles, called carbon nanodots, can be readily prepared using hydrothermal microwave-assisted synthesis and easily purified. These particles, with a mean particle size around 3 nm, are highly soluble in water and display mirror-image profile both in the UV-Vis and in the infrared regions, as detected by electronic and vibrational circular dichroism, respectively. Finally, the nanoparticles are used as templates for the formation of chiral supramolecular porphyrin assemblies, showing that it is possible to use and transfer the chiral information. This simple (and effective) methodology opens up exciting opportunities for developing a variety of chiral composite materials and applications.

Optical Aggregation of Gold Nanoparticles for SERS Detection of Proteins and Toxins in Liquid Environment: Towards Ultrasensitive and Selective Detection.

Optical forces are used to aggregate plasmonic nanoparticles and create SERS-active hot spots in liquid. When biomolecules are added to the nanoparticles, high sensitivity SERS detection can be accomplished. Here, we pursue studies on Bovine Serum Albumin (BSA) detection, investigating the BSA-nanorod aggregations in a range from 100 µM to 50 nM by combining light scattering, plasmon resonance and SERS, and correlating the SERS signal with the concentration. Experimental data are fitted with a simple model describing the optical aggregation process. We show that BSA-nanorod complexes can be optically printed on non-functionalized glass surfaces, designing custom patterns stable with time. Furthermore, we demonstrate that this methodology can be used to detect catalase and hemoglobin, two Raman resonant biomolecules, at concentrations of 10 nM and 1 pM, respectively, i.e., well beyond the limit of detection of BSA. Finally, we show that nanorods functionalized with specific aptamers can be used to capture and detect Ochratoxin A, a fungal toxin found in food commodities and wine. This experiment represents the first step towards the addition of molecular specificity to this novel biosensor strategy.

Non-invasive optical method for real-time assessment of intracorneal riboflavin concentration and efficacy of corneal cross-linking.

Keratoconus is the primary cause of corneal transplantation in young adults worldwide. Riboflavin/UV-A corneal cross-linking may effectively halt the progression of keratoconus if an adequate amount of riboflavin enriches the corneal stroma and is photo-oxidated by UV-A light for generating additional cross-linking bonds between stromal proteins and strengthening the biomechanics of the weakened cornea. Here we reported an UV-A theranostic prototype device for performing corneal cross-linking with the ability to assess corneal intrastromal concentration of riboflavin and to estimate treatment efficacy in real time. Seventeen human donor corneas were treated according to the conventional riboflavin/UV-A corneal cross-linking protocol. Ten of these tissues were probed with atomic force microscopy in order to correlate the intrastromal riboflavin concentration recorded during treatment with the increase in elastic modulus of the anterior corneal stroma. The intrastromal riboflavin concentration and its consumption during UV-A irradiation of the cornea were highly significantly correlated (R = 0.79; P = .03) with the treatment-induced stromal stiffening effect. The present study showed an ophthalmic device that provided an innovative, non-invasive, real-time monitoring solution for estimating corneal cross-linking treatment efficacy on a personalized basis.

Assessment of trans-scleral iontophoresis delivery of lutein to the human retina.

The efficacy of novel scleral iontophoresis device for in situ delivery of lutein to the human retina was assessed by Resonance Raman spectroscopy (RRS) technique. Eight human donor eye globes were used for experiments, 6 of which underwent trans-scleral iontophoresis delivery of lutein and the other 2 were used as controls. The scleral iontophoresis applicator was filled with liposome-enriched 0.1% lutein solution and the generator's current was set at 2.5 mA and delivered for 4 min. A custom RRS setup was used for detecting lutein in the inner sclera, choroid, retinal periphery and macula of treated samples and controls. Forty minutes after iontophoresis, the inner sclera, choroid and retinal periphery were greatly enriched with lutein (P < .05); no lutein was found in the same ocular regions of non-treated samples. In the same period, the average concentration of lutein in the macula (4.8 ± 1.7 ng/mm2 ) of treated samples was 1.3 times greater than controls (3.7 ± 1.0 ng/mm2 ; P = .4). Scleral iontophoresis was shown to be effective in delivering lutein to the human retina. Future studies will aim at assessing if this therapeutic strategy is valuable to enrich the macular pigment in human subjects.

Ring/Chain Morphology Control in Overall-Neutral, Internally Ion-Paired Supramolecular Polymers.

The self-assembly of internally ion-paired, neutral AA/BB-type supramolecular polymers composed of complementary di-ionizable homoditopic pairs of monomers is reported. Host-to-guest double-proton transfer mediates the recognition between bis-calix[5]arenedicarboxylic acids and α,ω-diaminoalkanes to yield cyclic, doughnut-shaped assemblies with morphologies (i.e., cyclic vs. linear) that can be controlled by means of external chemical stimuli. The behavior of these intriguing aggregates, both in solution and on surfaces, has been investigated by a combination of 1 H and DOSY NMR spectroscopy, light-scattering, and atomic force microscopy.

Assessment of stromal riboflavin concentration-depth profile in nanotechnology-based transepithelial corneal crosslinking.

PURPOSE: To determine the intrastromal concentration of riboflavin in nanotechnology-based transepithelial corneal crosslinking. SETTING: Consiglio Nazionale delle Ricerche, Messina, Italy. DESIGN: Experimental study. METHODS: Six human donor sclerocorneal tissues were used to evaluate penetration of nanotechnology-based riboflavin 0.1% solution in the stroma through the intact epithelium. Three additional tissues were deepithelialized and soaked with dextran 20.0%-enriched riboflavin 0.1% solution for 30 minutes. After corneal soaking with riboflavin, all tissues were irradiated using a 10 mW/cm2 device for 9 minutes. Two-photon emission fluorescence (TPEF) axial scanning measurements were collected in all specimens before treatment and immediately after corneal soaking with riboflavin and ultraviolet-A (UVA) irradiation of the cornea. The absorbance spectra of each tissue were collected at the same time intervals. The TPEF signals and absorbance spectra were used to calculate the concentration-depth profile of riboflavin in the corneal stroma during treatments. RESULTS: The mean stromal riboflavin concentration was 0.008% ± 0.003% (SD) and 0.017% ± 0.001% after transepithelial soaking with the nanotechnology-based solution and standard soaking, respectively (P = .001). After UVA irradiation of the cornea, the mean consumption of riboflavin was 52% ± 13% and 67% ± 2% in the study group and control group, respectively (P < .01). CONCLUSIONS: The nanotechnology-based platform was effective in enriching the anterior stroma with riboflavin through the intact epithelium, although the riboflavin concentration-depth profile rapidly decreased across the mid and posterior stroma. The treatment-induced stiffening effect on the corneal stroma was not assessed in this study.

SERS detection of Biomolecules at Physiological pH via aggregation of Gold Nanorods mediated by Optical Forces and Plasmonic Heating.

Strategies for in-liquid molecular detection via Surface Enhanced Raman Scattering (SERS) are currently based on chemically-driven aggregation or optical trapping of metal nanoparticles in presence of the target molecules. Such strategies allow the formation of SERS-active clusters that efficiently embed the molecule at the "hot spots" of the nanoparticles and enhance its Raman scattering by orders of magnitude. Here we report on a novel scheme that exploits the radiation pressure to locally push gold nanorods and induce their aggregation in buffered solutions of biomolecules, achieving biomolecular SERS detection at almost neutral pH. The sensor is applied to detect non-resonant amino acids and proteins, namely Phenylalanine (Phe), Bovine Serum Albumin (BSA) and Lysozyme (Lys), reaching detection limits in the µg/mL range. Being a chemical free and contactless technique, our methodology is easy to implement, fast to operate, needs small sample volumes and has potential for integration in microfluidic circuits for biomarkers detection.

All-Optical Method to Assess Stromal Concentration of Riboflavin in Conventional and Accelerated UV-A Irradiation of the Human Cornea.

PURPOSE: We investigated the concentration of riboflavin in human donor corneas during corneal cross-linking using two-photon optical microscopy and spectrophotometry. METHODS: Eight corneal tissues were de-epithelialized and soaked with 20% dextran-enriched 0.1% riboflavin solution for 30 minutes. After stromal soaking, three tissues were irradiated using a 3 mW/cm2 UV-A device for 30 minutes and three tissues irradiated using a 10 mW/cm2 device for 9 minutes. Two additional tissues were used as positive controls. A Ti:sapphire laser at 810 nm was used to perform two-photon emission fluorescence (TPEF) and second harmonic generation axial scanning measurements in all specimens before and after stromal soaking and after UV-A irradiation. In addition, spectrophotometry was used to collect the absorbance spectra of each tissue at the same time intervals. Analysis of the absorbance spectra and TPEF signals provided measures of the concentration depth profile of riboflavin in corneal stroma. RESULTS: After stromal soaking, the average peak concentration of riboflavin (0.020% ± 0.001%) was found between a stromal depth of 100 and 250 µm; the concentration of riboflavin was almost constant up to 320 ± 53 µm depth, then decreased toward the endothelium, though riboflavin was still enriched in the posterior stroma (0.016%% ± 0.001%). After conventional and accelerated UV-A irradiation, the concentration of riboflavin decreased uniformly 87% ± 2% and 67% ± 3% (P < 0.001), respectively. CONCLUSIONS: The combined use of two-photon optical microscopy and spectrophotometry provides relevant information for investigating the concentration of riboflavin in corneal stroma. The method can assist with the assessment of novel riboflavin formulations and different UV-A irradiation protocols.

Ultraviolet A: Visible spectral absorbance of the human cornea after transepithelial soaking with dextran-enriched and dextran-free riboflavin 0.1% ophthalmic solutions.

PURPOSE: To evaluate the stromal concentration of 2 commercially available transepithelial riboflavin 0.1% solutions in human donor corneas with the use of spectrophotometry. SETTING: University of Calabria, Rende, Italy. DESIGN: Experimental study. METHODS: The absorbance spectra of 12 corneal tissues were measured in the 330 to 700 nm wavelength range using a purpose-designed spectrophotometry setup before and after transepithelial corneal soaking with a 15% dextran-enriched riboflavin 0.1% solution (n = 6) or a hypotonic dextran-free riboflavin 0.1% solution (n = 6). Both ophthalmic solutions contained ethylenediaminetetraacetic acid and trometamol as enhancers. In addition, 4 deepithelialized corneal tissues underwent stromal soaking with a 20% dextran-enriched riboflavin 0.1% solution and were used as controls. All the riboflavin solutions were applied topically for 30 minutes. The stromal concentration of riboflavin was quantified by analysis of absorbance spectra of the cornea collected before and after application of each solution. RESULTS: The mean stromal riboflavin concentration was 0.012% ± 0.003% (SD), 0.0005% ± 0.0003% (P < .001), and 0.004% ± 0.001% (P < .01) in tissues soaked with 20% dextran-enriched, 15% dextran-enriched, and hypotonic dextran-free solutions, respectively. The difference of stromal riboflavin concentration between the 2 transepithelial solutions was statistically significant (P < .01). CONCLUSIONS: Dextran-enriched solutions required complete corneal deepithelialization to permit effective stromal soaking with riboflavin. Nevertheless, riboflavin in hypotonic dextran-free solution with enhancers permeates across stroma through an intact epithelium. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

New Evidence about the Spontaneous Symmetry Breaking: Action of an Asymmetric Weak Heat Source.

In the present study, we show how, in a stagnant water solution of uncharged aggregated achiral porphyrin-based molecules, a mirror-symmetry breaking (SB) can be induced and controlled by means of a weak asymmetric thermal gradient. In particular, it is shown that the optical activity of the aggregate porphyrin solution can be generated and reversed, in sign, only acting on the thermal ramp direction (heating or cooling). In order to avoid data misinterpretation, the aggregate structure modifications with the temperature change and the linear dichroism contribution to circular dichroism spectra were evaluated. A model simulation, using a finite element analysis approach describing the thermal flows, shows that small thermal gradients are able to give rise to asymmetric heat flow. The results reported here can be considered new evidence about the spontaneous symmetry breaking phenomenon induced by very weak forces having an important role in the natural chiral selective processes.

Hydrodynamic and Thermophoretic Effects on the Supramolecular Chirality of Pyrene-Derived Nanosheets.

Chiroptical properties of two-dimensional (2D) supramolecular assemblies (nanosheets) of achiral, charged pyrene trimers (Py3 ) are rendered chiral by asymmetric physical perturbations. Chiral stimuli in a cuvette can originate either from controlled temperature gradients or by very gentle stirring. The chiroptical activity strongly depends on the degree of supramolecular order of the nanosheets, which is easily controlled by the method of preparation. The high degree of structural order ensures strong cooperative effects within the aggregates, rendering them more susceptible to external stimuli. The samples prepared by using slow thermal annealing protocols are both CD and LD active (in stagnant and stirred solutions), whereas for isothermally aged samples chiroptical activity was in all cases undetectable. In the case of temperature gradients, the optical activity of 2D assemblies could be recorded for a stagnant solution due to migration of the aggregates from the hottest to the coldest regions of the system. However, a considerably stronger exciton coupling, coinciding with the J-band of the interacting pyrenes, is developed upon subtle vortexing (0.5 Hz, 30 rpm) of the aqueous solution of the nanosheets. The sign of the exciton coupling is inverted upon switching between clockwise and counter-clockwise rotation. The supramolecular chirality is evidenced by the appearance of CD activity. To exclude artefacts from proper CD spectra, the contribution from LD to the observed CD was determined. The data suggest that the aggregates experience asymmetrical deformation and alignment effects because of the presence of chiral flows.

Self-assembly of amphiphilic anionic calix[4]arenes and encapsulation of poorly soluble naproxen and flurbiprofen.

Supramolecular aggregates formed through the association of an amphiphilic tetra-O-butylsulfonate calix[4]arene 1 were investigated in aqueous solution by a combination of different techniques (NMR, DLS and AFM). The ability of the micellar aggregates of calixarene 1 to increase the solubility of poorly water-soluble drugs was studied.

Hierarchical effect behind the supramolecular chirality of silver(I)-cysteine coordination polymers.

Cysteine is a sulfur-containing amino acid that easily coordinates to soft metal ions and grafts to noble metal surfaces. Recently, chiroptical activity of Ag(+)/cysteine coordination polymers has been widely studied, while, on the other hand, the appearance of a plasmon-enhanced circular dichroic signal (PECD) at the plasmonic spectral region (λ > 400 nm) has been observed for AgNPs capped with chiral sulfur-containing amino acids. These two events are both potentially exploited for sensing applications. However, the presence of Ag(+) ions in AgNP colloidal solution deals with the competition of cysteine grafting at the metal NP surface and/or metal ion coordination. Herein we demonstrate that the chiroptical activity observed by adding cysteine to AgNP colloids prepared by pulsed laser ablation in liquids (PLAL) is mainly related to the formation of CD-active Ag(+)/cysteine supramolecular polymers. The strict correlation between supramolecular chirality and hierarchical effects, driven by different chemical environments experienced by cysteine when different titration modalities are used, is pivotal to validate cysteine as a fast and reliable probe to characterize the surface oxidation of AgNPs prepared by pulsed laser ablation in liquids by varying the laser wavelengths.

Control of the structural stability of α-crystallin under thermal and chemical stress: the role of carnosine.

The structural properties of α-crystallin, the major protein of the eye lens of mammals, in aqueous solution are investigated by means of small angle X-ray and dynamic light scattering. The research interest is devoted in particular to the effect of carnosine in protecting the protein under stress conditions, like temperature increase and presence of denaturant (guanidinium-HCl). The results suggest that carnosine interacts, through mechanisms involving hydrophobic interactions, with α-crystallin and avoids the structural changes in the quaternary structure induced by thermal and chemical stress. It is also shown that, if mediated by carnosine, the self-aggregation of α-crystallin induced by the denaturant at higher temperature can be controlled and even partially reversed. Therefore, carnosine is effective in preserving the structural integrity of the protein, suggesting the possibility of new strategies of intervention for preventing or treating pathologies related to protein aggregation, like cataracts.