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Verderio P.,University of Milan Bicocca | Avvakumova S.,University of Milan Bicocca | Avvakumova S.,University of Milan | Alessio G.,University of Milan | And 8 more authors.
Advanced Healthcare Materials | Year: 2014

Understanding the behavior of multifunctional colloidal nanoparticles capable of biomolecular targeting remains a fascinating challenge in materials science with dramatic implications in view of a possible clinical translation. In several circumstances, assumptions on structure-activity relationships have failed in determining the expected responses of these complex systems in a biological environment. The present Review depicts the most recent advances about colloidal nanoparticles designed for use as tools for cellular nanobiotechnology, in particular, for the preferential transport through different target compartments, including cell membrane, cytoplasm, mitochondria, and nucleus. Besides the conventional entry mechanisms based on crossing the cellular membrane, an insight into modern physical approaches to quantitatively deliver nanomaterials inside cells, such as microinjection and electro-poration, is provided. Recent hypotheses on how the nanoparticle structure and functionalization may affect the interactions at the nano-bio interface, which in turn mediate the nanoparticle internalization routes, are highlighted. In addition, some hurdles when this small interface faces the physiological environment and how this phenomenon can turn into different unexpected responses, are discussed. Finally, possible future developments oriented to synergistically tailor biological and chemical properties of nanoconjugates to improve the control over nanoparticle transport, which could open new scenarios in the field of nanomedicine, are addressed. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Mehn D.,Laboratory of Nanomedicine and Clinical Biophotonics | Morasso C.,Laboratory of Nanomedicine and Clinical Biophotonics | Vanna R.,Laboratory of Nanomedicine and Clinical Biophotonics | Bedoni M.,Laboratory of Nanomedicine and Clinical Biophotonics | And 2 more authors.
Vibrational Spectroscopy | Year: 2013

Paper-based SERS active substrates were prepared adsorbing spherical and star-shaped gold nanoparticles on a standard filter paper support. Besides the deposition conditions, morphological parameters of the particles were found to strongly affect the enhancer properties of the substrates. The developed substrate was tested regarding surface homogeneity as well as in the quantitative analysis of malachite green, - a well documented Raman reporter dye - and proved to be capable also to detect the oxidation products of apomorphine, a well-known drug molecule used in Parkinson's disease. This material is simple to prepare, easy to handle and dispose and as such it could be a perfect target for further development of a new family of mass-produced, cheap solid SERS substrates. © 2013 Elsevier B.V.

Verderio P.,University of Milan Bicocca | Verderio P.,Nerviano Medical science S.r.l | Pandolfi L.,University of Milan Bicocca | Pandolfi L.,University of Milan | And 10 more authors.
Molecular Pharmaceutics | Year: 2014

Among polymeric nanoparticles designed for cancer therapy, PLGA nanoparticles have become one of the most popular polymeric devices for chemotherapeutic-based nanoformulations against several kinds of malignant diseases. Promising properties, including long-circulation time, enhanced tumor localization, interference with "multidrug" resistance effects, and environmental biodegradability, often result in an improvement of the drug bioavailability and effectiveness. In the present work, we have synthesized 1,7-bis(3,4-dimethoxyphenyl)-5-hydroxyhepta-1,4,6-trien-3-one (ASC-J9) and developed uniform ASC-J9-loaded PLGA nanoparticles of about 120 nm, which have been prepared by a single-emulsion process. Structural and morphological features of the nanoformulation were analyzed, followed by an accurate evaluation of the in vitro drug release kinetics, which exhibited Fickian law diffusion over 10 days. The intracellular degradation of ASC-J9-bearing nanoparticles within estrogen-dependent MCF-7 breast cancer cells was correlated to a time- and dose-dependent activity of the released drug. A cellular growth inhibition associated with a specific cell cycle G2/M blocking effect caused by ASC-J9 release inside the cytosol allowed us to put forward a hypothesis on the action mechanism of this nanosystem, which led to the final cell apoptosis. Our study was accomplished using Annexin V-based cell death analysis, MTT assessment of proliferation, radical scavenging activity, and intracellular ROS evaluation. Moreover, the intracellular localization of nanoformulated ASC-J9 was confirmed by a Raman optical imaging experiment designed ad hoc. PLGA nanoparticles and ASC-J9 proved also to be safe for a healthy embryo fibroblast cell line (3T3-L1), suggesting a possible clinical translation of this potential nanochemotherapeutic to expand the inherently poor bioavailability of hydrophobic ASC-J9 that could be proposed for the treatment of malignant breast cancer. © 2014 American Chemical Society.

Morasso C.,Laboratory of Nanomedicine and Clinical Biophotonics | Mehn D.,Laboratory of Nanomedicine and Clinical Biophotonics | Vanna R.,Laboratory of Nanomedicine and Clinical Biophotonics | Bedoni M.,Laboratory of Nanomedicine and Clinical Biophotonics | And 5 more authors.
Materials Chemistry and Physics | Year: 2014

In this paper we present a new protocol for the synthesis of Star-Like Gold Nanoparticles (SGNs) by a simple one-step, room temperature procedure not involving the use of seeds or surfactants, that can be performed in seconds in any laboratory without the need of special technologies. These particles exhibited excellent properties for Surface Enhanced Raman Spectroscopy (SERS) and, when compared with spherical nanoparticles with similar size and concentration, showed enhancing factors from 10 to 50 times higher depending on the dye and on the wavelength employed. SGNs could be used directly in suspension as single, non-aggregating particles and were shown to be active in a remarkably broad range of the light spectrum from green to near infrared. Moreover, SGNs were adsorbed on the surface of a silicon slide to prepare SERS active solid substrate. Despite the fact that the surface of the solid substrate was not perfectly homogeneous, the signals recorded from different positions acquired through DuoScan averaging mode show excellent reproducibility, demonstrating how this simple and cheap protocol can be applied in order to generate reliable and homogeneous SERS substrates. © 2013 Elsevier B.V. All rights reserved.

Vanna R.,Laboratory of Nanomedicine and Clinical Biophotonics | Vanna R.,University of Twente | Tresoldi C.,San Raffaele Scientific Institute | Ronchi P.,San Raffaele Scientific Institute | And 8 more authors.
Progress in Biomedical Optics and Imaging - Proceedings of SPIE | Year: 2014

Acute myeloid leukemia (AML) is a proliferative neoplasm, that if not properly treated can rapidly cause a fatal outcome. The diagnosis of AML is challenging and the first diagnostic step is the count of the percentage of blasts (immature cells) in bone marrow and blood sample, and their morphological characterization. This evaluation is still performed manually with a bright field light microscope. Here we report results of a study applying Raman spectroscopy for analysis of samples from two patients affected by two AML subtypes characterized by a different maturation stage in the neutrophilic lineage. Ten representative cells per sample were selected and analyzed with high-resolution confocal Raman microscopy by scanning 64x64 (4096) points in a confocal layer through the volume of the whole cell. The average spectrum of each cell was then used to obtain a highly reproducible mean fingerprint of the two different AML subtypes. We demonstrate that Raman spectroscopy efficiently distinguishes these different AML subtypes. The molecular interpretation of the substantial differences between the subtypes is related to granulocytic enzymes (e.g. myeloperoxidase and cytochrome b558), in agreement with different stages of maturation of the two considered AML subtypes. These results are promising for the development of a new, objective, automated and label-free Raman based methods for the diagnosis and first assessment of AML. © 2014 SPIE.

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