Mariarosaria Tortora

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.

Nanomedicines for antimicrobial interventions

The development of new antimicrobial therapeutic tools addresses the emergence of multidrug-resistant micro-organisms or clones and the need for more effective antimicrobial strategies. Overcoming the hurdles in providing early diagnosis and intervention on hard-to-reach and/or resting bacteria (i.e. biofilm-embedded cells) represents a challenging task. In this review, we identify a set of organic, inorganic, and hybrid materials that might be used for prevention and control of healthcare-associated infections. We report the current knowledge on nano- and microparticle-based antimicrobial agents and describe the possible mode of their action.

Molecular properties of lysozyme-microbubbles: towards the protein and nucleic acid delivery

Microbubbles (MBs) have specific acoustic properties that make them useful as contrast agents in ultrasound imaging. The use of the MBs in clinical practice led to the development of more sensitive imaging techniques both in cardiology and radiology. Protein-MBs are typically obtained by dispersing a gas phase in the protein solution and the protein deposited/cross-linked on the gas–liquid interface stabilizes the gas core. Innovative applications of protein-MBs prompt the investigation on the properties of MBs obtained using different proteins that are able to confer them specific properties and functionality. Recently, we have synthesized stable air-filled lysozyme-MBs (LysMBs) using high-intensity ultrasound-induced emulsification of a partly reduced lysozyme in aqueous solutions. The stability of LysMBs suspension allows for post-synthetic modification of MBs surface. In the present work, the protein folded state and the biodegradability property of LysMBs were investigated by limited proteolysis. Moreover, LysMBs were coated and functionalized with a number of biomacromolecules (proteins, polysaccharides, nucleic acids). Remarkably, LysMBs show a high DNA-binding ability and protective effects of the nucleic acids from nucleases and, further, the ability to transform the bacteria cells. These results highlight on the possibility of using LysMBs for delivery of proteins and nucleic acids in prophylactic and therapeutic applications.

Redox-active Microcapsules as Drug Delivery System in Breast Cancer Cells and Spheroids

The purpose of our study was to develop new delivery systems for drugs effective against breast cancer by using biodegradable and biocompatible capsules. Redox-active microcapsules based on thiolated polymethacrylic acid (PMA) were employed. The interaction of these PMASH capsules with breast cancer cells and the mechanism of their internalization was investigated. PMASH biocompatibility was evaluated by MTT assay. To analyze their potential as drug carrier, we incorporated doxorubicin into the capsules. Confocal microscopy observations showed the presence of capsules inside the cells. Although some drug molecules still appeared co-localized with PMASH capsules, strong doxorubicin fluorescence was observed both in the cytoplasm and nucleus, indicating the disassembling of the capsule into PMASH-drug conjugate after internalization. These results were confirmed by both flow cytometry (time course of capsule uptake) and scanning electron microscopy. PMASH capsules were also internalized in 3D cell structures (spheroids) suggesting their potential use as drug delivery system for treatment of human diseases.

Design of Novel Polymer Shelled Ultrasound Contrast Agents: Towards an Ultrasound Triggered Drug Delivery

Ultrasound contrast agents (UCA) have been used for years in the clinical field, for applications such as blood pool enhancement, characterization of liver lesions or perfusion imaging [12, 13, 14, 15, 20]. The contrast agents are generally in the form of spherical voids or cavities filled by a gas, called microbubbles (MB). MBs are stabilized by a coating material such as phospholipids, surfactants, denatured human serum albumin or synthetic polymers. As gas is less dense than liquids or solids, sound travels more slowly in gas than it does in liquid. The difference in the sound speed in the microbubbles creates an acoustic mismatch between tissue and blood surrounding a microbubble, making it an efficient reflector of ultrasound energy. The ability to produce strong signals from microbubbles depends on the stability of the gas particle. Stability has been enhanced by varying the gas composition (air, nitrogen, sulphur hexafluoride, perfluorocarbons) and/or by chemical modification of the microbubble shell [20]. As microbubbles stay in the vascular space and have a behavior similar to red blood cells in the microcirculation, they can be used as intravascular tracers. The possibility to introduce a targeting ligand for a specific interaction with its receptor onto the microbubble shell can be a way to selectively accumulate the contrast agent in the deseased region. Moreover, targeted MBs can be used to carry different drugs to the desired sites and release them after their destruction/cavitation with ultrasound radiation or by chemical/enzymatic cleavage.

Polymer Based Biointerfaces: A Case Study on Devices for Theranostics and Tissue Engineering

In the design of new devices supporting biomedical applications, the focus on the processes occurring at the interface with cells is a major issue. In this context, our interest on the formulation of new biomaterials and on the synthesis of next-generation ultrasound contrast agents (UCAs) lead us to the in vitro study to assess the biocompatibility of these novel devices. As UCAs are designed for parenteral administration, the time response of the primary immune system, mainly macrophages, should be addressed.

A combined approach for predicting the cytotoxic effect of drug-nanoaggregates

We present a combined spectroscopic and computational approach aimed to elucidate the mechanism of formation and activity of etoposide nanoaggregates upon release from dextran-etoposide conjugates. Etoposide is an anticancer drug that inhibits cell growth by blocking Topoisomerase II, the key enzyme involved in re-ligation of the DNA chains during the replication process. In silico and spectroscopic analysis indicate that released etoposide nanoaggregates have a different structure, stability, and bioactivity, which depend on the pH experienced during the release. Molecular dynamics simulation and in silico docking of etoposide dimers suggest that the aggregation phenomena inhibit etoposide bioactivity, yet without drastically preventing Topoisomerase II binding. We correlated the diminished cytotoxic activity exerted by dextran-etoposide conjugates on the A549 lung cancer cells, compared to the free drug, to the formation and stability of drug nanoaggregates.

Drug delivery system and breast cancer cells

Recently, nanomedicine has received increasing attention for its ability to improve the efficacy of cancer therapeutics. Nanosized polymer therapeutic agents offer the advantage of prolonged circulation in the blood stream, targeting to specific sites, improved efficacy and reduced side effects. In this way, local, controlled delivery of the drug will be achieved with the advantage of a high concentration of drug release at the target site while keeping the systemic concentration of the drug low, thus reducing side effects due to bioaccumulation. Various drug delivery systems such as nanoparticles, liposomes, microparticles and implants have been demonstrated to significantly enhance the preventive/therapeutic efficacy of many drugs by increasing their bioavailability and targetability. As these carriers significantly increase the therapeutic effect of drugs, their administration would become less cost effective in the near future. The purpose of our research work is to develop a delivery system for breas...