Francesca Cavalieri

Poly(vinyl alcohol) as versatile biomaterial for potential biomedical applications

In this paper, we present some new case examples where the chemical versatility of poly (vinyl alcohol) (PVA) can be used for potential biomedical applications. PVA, the polymeric material used for designing new nanostructured devices, is water soluble, biocompatible and has excellent physical properties. We point out the possibility of obtaining wall-to-wall chemical hydrogels as well as microgels without diminishing the biocompatibility available in the starting PVA material. Injectability is another important factor to take into account in controlled drug delivery for gene therapy. In this respect, in this paper, established and more innovative methods are prospected in order to obtain particles with dimensions suitable for these applications.

Ultrasonic synthesis of stable, functional lysozyme microbubbles.

High-intensity ultrasound induces emulsification and cross-linking of protein molecules in aqueous medium. The stability and the functionality of the resultant protein-coated microbubbles are crucial in many of their applications. For example, the stability of drug-loaded microbubbles should be sufficiently long enough, in vivo, so that they can be ruptured only at specific sites for release of the drugs. In this study, we report the synthesis of stable and functional microbubbles, coated with chemically reduced lysozyme, using high-intensity ultrasound in aqueous solution. In the absence of chemical reduction, stable microbubbles were not produced with native lysozyme, indicating the importance of free -SH functional groups for protein cross-linking. The degree of cross-linking between lysozyme molecules was controlled by manipulating both the extent of chemical reduction of the intramolecular disulfide bonds and sonication time. The lysozyme-coated microbubbles are stable for several months and retain the enzymatic (antimicrobial) activity of lysozyme. The layer-by-layer (LbL) deposition of polyelectrolytes onto the protein-shell air-core template has been used as a versatile procedure to modify the surface properties of the microbubbles, indicating the possibility of adsorbing potential drugs and/or biolabels on the surface of these microbubbles for therapeutic and diagnostic applications.

Redox-Active Polymer Microcapsules for the Delivery of a Survivin-Specific siRNA in Prostate Cancer Cells

In this report, we describe the delivery of small interfering RNA (siRNA) using LbL-assembled microcapsules. The microcapsules are based on negatively charged poly(methacrylic acid) nanometer thin films containing cross-linking disulfide bonds. One system is polycation-free and another contains polylysine for siRNA complexation in the microcapsule void. When microcapsules containing a siRNA targeting survivin were delivered to PC-3 prostate cancer cells, a significant inhibition of the expression of the antiapoptotic protein was observed. However, down-regulation of survivin was also observed in PC-3 cells exposed to microcapsules embedded with a scrambled siRNA as well as in cells treated with empty microcapsules. These findings indicate a capsule-dependent off-target effect, which is supported by a reduction in the expression of other survivin-unrelated proteins. The microcapsules and their polymeric constituents do not affect cell proliferation, as determined by a metabolic assay, even after 4 days of exposure. In addition, in PC-3 cells exposed to microcapsules, we observed a marked accumulation of LC3b, a marker related to autophagy (i.e., self-digestion), a degradation pathway involved in the maintenance of cell homeostasis in response to different stresses. This evidence suggests that empty microcapsules can induce a perturbation of the intracellular environment, which causes the activation of a cell safeguard mechanism that may limit the therapeutic effect of the microcapsules in tumor cells.

Ultrasound Driven Assembly of Lignin into Microcapsules for Storage and Delivery of Hydrophobic Molecules

Oil-filled microcapsules of kraft lignin were synthesized by first creating an oil in water emulsion followed by a high-intensity, ultrasound-assisted cross-linking of lignin at the water/oil interface. The rationale behind our approach is based on promoting documented lignin hydrophobic interactions within the oil phase, followed by locking the resulting spherical microsystems by covalent cross-linking using a high intensity ultrasound treatment. As further evidence in support of our rationale, confocal and optical microscopies demonstrated the uniformly spherical morphology of the created lignin microparticles. The detailed elucidation of the cross-linking processes was carried out using gel permeation chromatography (GPC) and quantitative (31)P NMR analyses. The ability of lignin microcapsules to incorporate and release Coumarin-6 was evaluated in detail. In vitro studies and confocal laser scanning microscopy analysis were carried out to assess the internalization of capsules into Chinese hamster ovary (CHO) cells. This part of our work demonstrated that the lignin microcapsules are not cytotoxic and readily incorporated in the CHO cells.

Novel PVA-based hydrogel microparticles for doxorubicin delivery.

Micro- and nanoparticles are considered suitable drug delivery systems for their unique features, such as a large surface to volume ratio, and for the possibility to tune their size and hydrophobicity. A polymer/polymer/water emulsion method was used for producing a chemically cross-linked hydrogel made of poly(vinyl alcohol) and of poly(methacrylate) moieties. Mesoscopic investigation of the microparticles was accomplished by laser scanning confocal microscopy. Dynamics of confined water within the gel meshes was studied by quasi-elastic incoherent neutron scattering. Succinoylation of these particles allowed an efficient loading with a maximum doxorubicin payload of about 50% (w/w) of dry microparticles. To evaluate the potentials of such a microdevice for drug delivery, LoVo colon cancer cells have been exposed to doxorubicin loaded microparticles to study the in vitro efficiency of the payload release and the consequent cytotoxic effect.

Development of composite materials by mechanochemical treatment of post-consumer plastic waste

Improvement of mechanical properties of recycled mixed plastic waste is one of the fundamental goals in any recycling process. However, polymer immiscibility makes the development of any effective reprocessing method difficult. In this work, a polymer milling process with liquid CO2 was applied to polymeric mixed waste, obtaining a powder material which was successfully utilized as a matrix for a new composite material. Developed materials have interesting mechanical properties and material performance can easily be improved. Investigations on selected mixtures of PP and PE clearly showed evidence of chemical compatibilization.

The Design of Multifunctional Microbubbles for Ultrasound Image-Guided Cancer Therapy

Gas-filled microbubbles are widely used in diagnostic imaging. Recent developments have greatly enhanced the potential use of microbubbles for both diagnostic and therapeutic applications. For the potential use of microbubbles in therapeutic applications, the chemical nature of the shell and its mechanical properties are crucial, and requires a tailored synthetic approach. This review describes methods of preparation, mechanism of action, in vitro and in vivo stability and structural/functional characterization of microbubbles. New mechanisms for ultrasound-enhanced local drug and gene delivery are reviewed. Different strategies used to target microbubbles to regions of disease and some of the recent experiences in ultrasound image-guided therapy are discussed.

High-energy mechanical alloying of thermoplastic polymers in carbon dioxide

Abstract High-energy ball milling was performed on low density polyethylene (LDPE) and isotactic polypropylene (iPP) as well as on 20/80 binary mixture of both polymers. Mechanical alloying was carried out at high pressure with carbon dioxide for a short period. The presence of CO2 avoids oxidative mechano-chemical degradation of polymers and enhances the effectiveness of the milling. The effects of the mechano-chemical treatment on the molecular and physical properties of both single polymers and blends of intrinsically incompatible polymers were explored by FTIR spectroscopy, thermal analysis, intrinsic viscosity determination and solvent fractionation. Structural changes on PP and PP/LDPE blend were observed and have a strong dependence on the milling time. Mechanical tests confirm an overall improvement in blend properties by mechanical alloying. Experimental evidences are presented to suggest that CO2 high-energy ball milling causes a self-compatibilization of the blend LDPE–iPP by breaking iPP polymer chains and allowing them to recombine with the neighboring LDPE chains.

New hydrogels based on carbohydrate and on carbohydrate-synthetic polymer networks

Abstract Novel synthetic routes recently explored in our laboratories in order to obtain crosslinked polymeric samples, comprising either chitosan or gellan as polysaccharidic components, are outlined. The former networks, in which different crosslinking partners have been used (i.e. oxidized cyclodextrin and telechelic poly(vinylalcohol)), can bear fixed positive charges while the latter are anionic. Zwitterionic networks formed by the template polymerization of acrylic acid onto chitosan with concomitant crosslinking (bisacrylamide) have also been prepared. Procedures adopted are simple and, once optimized, may lead to biocompatible hydrogels, with easily tunable physical properties, of potential interest in the biomedical area.

Tailoring the properties of ultrasonically synthesised microbubbles

Air-filled lysozyme microbubbles can be synthesized in an aqueous medium by emulsification followed by the cross-linking of protein molecules under high-intensity ultrasound. Here, we report on the tailoring of the properties of the ultrasonically synthesised microbubbles using new procedures. The efficiency of formation, size, size distribution and morphology of the microbubbles were controlled by manipulating the experimental conditions, namely, the sonication power and the length of sonication. An increase in the sonication time and power led to the formation of larger microbubbles with a broader size distribution. The microbubble shell thickness was found to decrease with an increase in the sonication power and time. Furthermore, a pulsed sonoluminescence technique was used to study the strength and stability of the microbubbles. The experimental results have shown that the effects of sonication time and power on the properties of the microbubbles are quite complex. A simple graphical matrix has been derived to obtain stable microbubbles with a narrow size distribution.