Valentina Villari

The intracellular effects of non-ionic amphiphilic cyclodextrin nanoparticles in the delivery of anticancer drugs

The aim of this study was to develop nanoparticles made of the amphiphilic cyclodextrin heptakis (2-O-oligo(ethyleneoxide)-6-hexadecylthio-)-beta-CD (SC16OH) entrapping docetaxel (Doc) and establish their in vivo potential. Doc-loaded SC16OH nanoparticles were prepared by the emulsion-solvent evaporation technique and fully characterized for size, zeta potential, amount of entrapped drug, release rate and degradation rate. Spherical vesicular nanoparticles displaying a hydrodynamic radius of about 95 nm which did not change upon storage as an aqueous dispersion, a negative zeta potential and entrapment efficiency of Doc very close to 100% were produced. DSC study highlighted the crystalline nature of SC16OH, unloaded and Doc-loaded SC16OH nanoparticles which resulted in their very slow dissolution during release stage and well-modulated release of entrapped Doc for about 8 weeks. Doc-loaded SC16OH nanoparticles were not hemolytic toward red blood cells as compared to a commercial Doc formulation (Taxotere) which shows a dose-dependent toxicity. After exposure of HEp-2 cells to equivalent doses of free Doc and Doc-loaded SC16OH nanoparticles, superior cell killing and cell damage were observed for nanoparticles. Finally, cell damage was attributed to aberrant mitosis which was found to be significantly higher for HEp-2 cells treated with Doc-loaded SC16OH nanoparticles as compared to free Doc likely due to the ability of nanoparticles to slowly release the drug allowing prolonged cell arrest in mitosis. Taken together, these results highlights a great potential of nanoparticles based on SC16OH in solid tumors therapy.

Scaling the Chirality in Porphyrin J-Nanoaggregates

Chiral porphyrin J-aggregates are confined into microemulsions, achieving a fine control of their size and optical properties. The quality of the circular dichroism signal exhibits a remarkable scaling on the dimension from the nano- up to mesoscopic regime.

Self-organizing functional materials via ionic self assembly: porphyrins H- and J-aggregates on synthetic chrysotile nanotubes.

Positively charged, synthetic chrysotile nanotubes act as inorganic tectons supporting H- and J-type aggregates of anionic porphyrins with markedly different optical properties, making these nanohybrid materials of interest for application in nanotechnology.

The puzzle of poly(ethylene oxide) aggregation in water: Experimental findings

Aqueous solutions of poly(ethylene oxide) were investigated using the ultrasonic technique, photon correlation spectroscopy (PCS) and nuclear magnetic resonance (NMR), in a wide range of molecular weight (from ethylene glycol to poly(ethylene oxide) 4 000 000 Da). Ultrasonic data reveal that the mixing process is not ideal and show that the polymer–water interaction strength increases with the polymerization degree. PCS and NMR, on the other hand, furnish a free particle diffusion coefficient which satisfies a unique scaling law from 8000 to 4 000 000 Da and demonstrates the good solvent nature of water. These experimental findings indicate that polymer–polymer aggregation processes are not an inherent property of these systems.

Anion-assisted supramolecular polymerization: from achiral AB-type monomers to chiral assemblies.

Control over the self-assembly process of monomeric species by functional group modulation is highly desirable in the context of supramolecular polymer design. These materials, unlike covalently linked polymers, consist of monomeric arrays held together by reversible and highly directional noncovalent bonds. Owing to the dynamic and reversible nature of noncovalent interactions, supramolecular polymers display unique topologies and unconventional properties (such as stimuli responsiveness and self-healing) and are thus becoming cutting-edge species in modern materials science. Multiple hydrogen bonds, metal–ligand coordination, and p–p stacking are, by far, the most common weak forces used for engineering supramolecular polymers. Recently, however, oligomeric and polymeric architectures based on host–guest inclusion complexes have started to become more and more popular. Within this research frame, we have recently described a pH-responsive aminododecyloxy-calix[5]arene derivative (C5-NH2) that, upon exposure to a variety of acids, selfassembles into linear oligomers. Protonation activates the two latent self-complementary binding sites of this heteroditopic monomer precursor (i.e. a preorganized cone-shaped p-rich calix[5]arene cavity and a linear alkylamine pendant chain) and, according to a well-established host–guest recognition pattern, which involves a concerted set of weak interactions (NH···O, CH–p, cation–p), supramolecular oligomer formation readily occurs. However, because of the intrinsically saline nature of the monomers used, the growth of these supramolecular assemblies was found to be aniondependent. More specifically, the looser the ion-pairing interactions between the ammonium monomer and its counterion, the higher the degree of polymerization observed. Although ion-pairing effects have been analyzed extensively in relation to simple one-to-one host–guest systems, to the best of our knowledge they have not yet been examined in the context of supramolecular polymers derived from charged monomers. Elegant examples of polymeric species derived from crown ethers, cryptands, cyclodextrins, cucurbiturils, calixarenes, 16] and resorcinarenes have been described, but in none of these instances—neither ABtype (self-complementary heteroditopic) 12a,b, 13a, 17a] nor AA/BB-type (complementary homoditopic) 15, 17b] systems—has the role of the counterion in the growth of the polymer or the tuning of the supramolecular properties been addressed. Drawing on our earlier investigations on the simultaneous complexation of cations and anions 18] and on the design of heteroditopic and heterotetratopic receptors in an attempt to override the drawback of ion-pairing effects in AB-type salt monomers, we have now incorporated an ancillary anion-binding site (namely a ureido moiety) into calix[5]arene C5-NH2 with the aim of facilitating salt dissociation and ultimately making polymer formation more efficient. In this communication we demonstrate that the addition of this anion-binding site to the monomer scaffold is beneficial to the supramolecular polymerization process and, most importantly, we show that modulation of the properties [*] Dr. C. Capici, Dr. G. Gattuso, Dr. A. Notti, Prof. M. F. Parisi Dipartimento di Chimica Organica e Biologica Universit di Messina Viale F. Stagno d’Alcontres 31, 98166 Messina (Italy) E-mail: ggattuso@unime.it mparisi@unime.it

Nanostructures of cationic amphiphilic cyclodextrin complexes with DNA.

Complexes of cationic amphiphilic cyclodextrins heptakis[2-(ω-amino-oligo(ethylene glycol))-6-deoxy-6-hexadecylthio]-β-cyclodextrin and heptakis[2-(ω-amino-oligo(ethylene glycol))-6-deoxy-6-dodecylthio]-β-cyclodextrin with DNA were examined by small-angle X-ray scattering and dynamic as well as electrophoretic light scattering. The first cyclodextrin forms bilayer vesicles in water, which, in the presence of calf thymus DNA, transform to a multilamellar complex. In this complex, the DNA lies between the two polar layers of the cyclodextrins protonated amino groups in alternation with the lipidic bilayers. The cyclodextrin with shorter lipid chains, in contrast, forms micelles in water, and electrostatic clustering of these about DNA does not affect their intrinsic structure. These results are relevant to the potential of such cyclodextrins in therapeutic gene delivery, showing that their self-assembly modes in isolation influence their complex formation with DNA and possibly their efficiency in promoting cell transfection.

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.

Experimental simulation of macromolecules in trehalose aqueous solutions: A photon correlation spectroscopy study

The protective effect of trehalose on biological membranes against freezing or dehydration has been the subject of many studies aimed to understand the reasons why some lower organisms, under stress conditions, synthesize trehalose. In this work we report the results of a study on Poly(Ethylene Oxide)/trehalose/water mixtures performed by Photon Correlation Spectroscopy. The chemical structure of the polymer, simpler than that of proteins and its helical conformation in water, constitute a useful starting point for understanding the more complex protein/trehalose/water interactions. In order to distinguish the different dynamics, trehalose and PEO have been studied separately in water, at different concentration and temperature values; then the ternary PEO/trehalose/water system has been investigated at different sugar amounts. The obtained findings support the “water-replacement” hypothesis, indicating that a direct polymer–trehalose interaction occurs. Furthermore, trehalose is shown to affect the swell...

Probing specific protein recognition by size-controlled glycosylated cyclodextrin nanoassemblies

The balance between hydrophobic and hydrophilic components in amphiphilic β-cyclodextrins, targeted by receptor specific groups (SC6CDGlc, SC6CDGal, SC16CDGlc, SC16CDGal), sensitively influences the structural properties of these systems. The different amphiphilic features of single cyclodextrins generate micellar aggregates and vesicles with an internal aqueous compartment able to encapsulate guests, such as rhodamine 6G. Small-angle light scattering (SAXS), cryo-TEM and AFM investigations describe the size and shape of these self-organized glycoligands. Recognition of the nanoassemblies by a specific receptor has effectively been demonstrated by means of time resolved fluorescence and is addressed in water by the morphological properties of cyclodextrin aggregates. Exclusively galactosylated thiohexyl-cyclodextrin binds specifically lectin from Pseudomonas aeruginosa. β-D-Galactose competes with galactosylated cyclodextrin aggregates by inhibiting lectin binding but does not affect the mesoscopic environment of the protein. The better selectivity of the less hydrophobic cyclodextrins towards lectin should probably be ascribed to the morphology (size and shape) of these cyclodextrin aggregates. The recognition properties of this particular cyclodextrin (SC6CDGal) are probably due to the presence of small micelles which interact more efficiently with the lectin binding site. The modulation of the hydrophobic–hydrophilic balance of the macrocycle labelled with targeting groups allows the design of “active” nanosized carriers for drug delivery.

Conformational distribution of poly(ethylene oxide) in molten phase and in aqueous solution by quasi-elastic and inelastic light scattering

We report on measurements performed by Raman scattering, PCS and ultrasonic velocity measurements on poly(ethylene oxide) both in the molten phase and in aqueous solution. Increasing the polymerization degree, m, the Raman analysis of the D-LAM (disordered longitudinal acoustic mode) spectral contribution to the pure polymers reveals a behaviour of the centre frequency and linewidth which has been connected with an oligomer-polymer transition occurring at . In aqueous solutions the frequency increase towards values corresponding to the crystal ones and the sharpening of the D-LAM spectral contribution indicate that the addition of water destroys the intermolecular interactions and stiffens the coil structure. In addition evidence of a more ordered conformation with respect to the melt phase is presented. The temperature analysis of the D-LAM band and of the hydrodynamic radius, evaluated by PCS, reveals that the solvent power of water increases up to , decreasing at higher temperature. Interpreted in conjunction with ultrasonic data, these apparently differing findings provide a coherent interpretative key capable of encompassing the structural properties of our systems. Finally the role played by inter- and intra-molecular interactions is discussed within the framework of current theories.