Cytocompatibility and Uptake of Halloysite Clay Nanotubes

Halloysite is aluminosilicate clay with hollow tubular structure of 50 nm external diameter and 15 nm diameter lumen. Halloysite biocompatibility study is important for its potential applications in polymer composites, bone implants, controlled drug delivery, and for protective coating (e.g., anticorrosion or antimolding). Halloysite nanotubes were added to different cell cultures for toxicity tests. Its fluorescence functionalization by aminopropyltriethosilane (APTES) and with fluorescently labeled polyelectrolyte layers allowed following halloysite uptake by the cells with confocal laser scanning microscopy (CLSM). Quantitative Trypan blue and MTT measurements performed with two neoplastic cell lines model systems as a function of the nanotubes concentration and incubation time indicate that halloysite exhibits a high level of biocompatibility and very low cytotoxicity, rendering it a good candidate for household materials and medicine. A combination of transmission electron microscopy (TEM), scanning electron microscopy (SEM), and scanning force microscopy (SFM) imaging techniques have been employed to elucidate the structure of halloysite nanotubes

Low-Intensity Light-Responsive Anticancer Activity of Platinum(II) Complex Nanocolloids on 2D and 3D In Vitro Cancer Cell Model

none8This study aimed to evaluate the therapeutic efficacy of low-intensity visible light responsive nanocolloids of a Pt-based drug using a 2D and three-dimensional (3D) in vitro cancer cell model. Biocompatible and biodegradable polymeric nanocolloids, obtained using the ultrasonication method coupled with Layer by Layer technology, were characterized in terms of size (100 ± 20 nm), physical stability, drug loading (78%), and photoactivation through spectroscopy studies. The in vitro biological effects were assessed in terms of efficacy, apoptosis induction, and DNA-Pt adducts formation. Biological experiments were performed both in dark and under visible light irradiation conditions, exploiting the complex photochemical properties. The light-stimuli responsive nanoformulation gave a significant enhancement in drug bioactivity. This allowed us to achieve satisfying results by using nanomolar drug concentration (50 nM), which was ineffective in darkness condition. Furthermore, our nanocolloids were validated in 3D in vitro spheroids using confocal microscopy and cytofluorimetric assay to compare their behavior on culture in 2D monolayers. The obtained results confirmed that these nanocolloids are promising tools for delivering Pt-based drugs.This study was supported by “Tecnopolo di Nanotecnologia e Fotonica per la Medicina di Precisione” (TECNOMED)-FISR/MIUR-CNR: delibera CIPE n.3449 del 7/08/2017, CUP: B83B17000010001; “Tecnopolo per la Medicina di precisione” (TecnoMed Puglia) - Regione Puglia: DGR n.2117 del 21/11/2018, CUP: B84I18000540002.openViviana Vergaro; Francesca Baldassarre; Federica De Castro; Danilo Migoni; Maria Michela Dell’Anna; Piero Mastrorilli; Francesco Paolo Fanizzi; Giuseppe CiccarellaVergaro, Viviana; Baldassarre, Francesca; DE CASTRO, Federica; Danilo, Migoni; Michela Dell’Anna, Maria; Mastrorilli, Piero; Fanizzi, Francesco Paolo; Ciccarella, Giusepp

Enhanced Bioactivity of Pomegranate Peel Extract following Controlled Release from CaCO3 Nanocrystals

none9Pomegranate peel extract is rich of interesting bioactive chemicals, principally phenolic compounds, which have shown antimicrobial, anticancer, and antioxidative properties. The aim of this work was to improve extract’ bioactivity through the adsorption on calcium carbonate nanocrystals. Nanocrystals revealed as efficient tools for extract adsorption reaching 50% of loading efficiency. Controlled release of the contained metabolites under acidic pH has been found, as it was confirmed by quantitative assay and qualitative study through NMR analysis. Specific functionality of inorganic nanocarriers could be also tuned by biopolymeric coating. The resulting coated nanoformulations showed a great antimicrobial activity against B. cinerea fungus preventing strawberries disease better than a commercial fungicide. Furthermore, nanoformulations demonstrated a good antiproliferative activity in neuroblastoma and breast cancer cells carrying out a higher cytotoxic effect respect to free extract, confirming a crucial role of nanocarriers. Finally, pomegranate peel extract showed a very high radical scavenging ability, equal to ascorbic acid. Antioxidant activity, measured also in intracellular environment, highlighted a protective action of extract-adsorbed nanocrystals twice than free extract, providing a possible application for new nutraceutical formulations.Regione Puglia, Project Research for Innovation (REFIN) “Sintesi di un sistema teranostico a base di nano-cellulosa per la detection e la cura dei tumori” Ministero dello Sviluppo Economico, bando “AGRIFOOD” D.M. 5 marzo 2018 Capo III. Prog. N. F/200060/01-03/X45 PERSEFONE - Punica granatum e Nanotecnologie: una value chain per la valorizzazione degli scarti e sottoprodotti finalizzata alla produzione di integratori alimentari e compost. PON Ricerca e Innovazione 2014-2020 - Avviso per la presentazione di progetti dei Ricerca Industriale e Sviluppo Sperimentale nelle 12 aree di specializzazione individuate dal PNR 2015 – 2020, di cui al Decreto Direttoriale MIUR del 13 luglio 2017, n. 1735 NanotEcnologie chiMiche green per la protEzione Sostenibile delle pIante (NEMESI) ARS01_01002 Area di Specializzazione “Chimica Verde” CUP: F36C18000180005, Ministero dell'Università e Ricerca. “Olivicoltura e difesa da Xylella fastidiosa e da insetti vettori in Italia - (Oli.Di.X.I.It)”, prot. Mipaaf n.0011485 del 05/04/2017 Ministry of Agricultural, Food and Forestry Policies.openFrancesca Baldassarre; Viviana Vergaro; Federica De Castro; Francesca Biondo; Gian Paolo Suranna; Paride Papadia; Francesco P. Fanizzi; Domenico Rongai; Giuseppe CiccarellaBaldassarre, Francesca; Vergaro, Viviana; DE CASTRO, Federica; Biondo, Francesca; Suranna, Gian Paolo; Papadia, Paride; Fanizzi, Francesco P.; Rongai, Domenico; Ciccarella, Giusepp

Sonication-Assisted Production of Fosetyl-Al Nanocrystals: Investigation of Human Toxicity and In Vitro Antibacterial Efficacy against Xylella Fastidiosa

none10Recently, there is a growing demand in sustainable phytopathogens control research. Nanotechnology provides several tools such as new pesticides formulations, antibacterial nanomaterials and smart delivery systems. Metal nano-oxides and different biopolymers have been exploited in order to develop nanopesticides which can offer a targeted solution minimizing side effects on environment and human health. This work proposed a nanotechnological approach to obtain a new formulation of systemic fungicide fosetyl-Al employing ultrasonication assisted production of water dispersible nanocrystals. Moreover, chitosan was applicated as a coating agent aiming a synergistic antimicrobial effect between biopolymer and fungicide. Fosetyl-Al nanocrystals have been characterized by morphological and physical-chemical analysis. Nanotoxicological investigation was carried out on human keratinocytes cells through cells viability test and ultrastructural analysis. In vitro planktonic growth, biofilm production and agar dilution assays have been conducted on twoXylella fastidiosasubspecies. Fosetyl-Al nanocrystals resulted very stable over time and less toxic respect to conventional formulation. Finally, chitosan-based fosetyl-Al nanocrystals showed an interesting antibacterial activity againstXylella fastidiosasubsp.paucaandXylella fastidiosasubsp.fastidiosa.Regione Puglia, Deliberazione della Giunta Regionale n. 1410 del 12/06/2015 “Linee guida per il parco della ricerca e sperimentazione finalizzata alla prevenzione e al contenimento del complesso del disseccamento rapido dell’olivo (CODIRO)” Project FATA- “Trattamenti fitoterapici innovativi a base di vettori di chitosano” MIPAAFT, Project Oli.Di.X.I.It-“OLIvicoltura e Difesa da Xylella fastidiosa e da Insetti vettori in Italia”, D.M. 23773 del 6/09/2017.openFrancesca Baldassarre; Giuseppe Tatulli; Viviana Vergaro; Stefania Mariano; Valeria Scala; Concetta Nobile; Nicoletta Pucci; Luciana Dini; Stefania Loreti; Giuseppe CiccarellaBaldassarre, Francesca; Tatulli, Giuseppe; Vergaro, Viviana; Mariano, Stefania; Scala, Valeria; Nobile, Concetta; Pucci, Nicoletta; Dini, Luciana; Loreti, Stefania; Ciccarella, Giusepp

Lapatinib/Paclitaxel polyelectrolyte nanocapsules for overcoming multidrug resistance in ovarian cancer.

The sonication-assisted layer-by-layer (SLBL) technology was developed to combine necessary factors for an efficient drug-delivery system: (i) control of nanocolloid size within 100 - 300 nm, (ii) high drug content (70% wt), (iii) shell biocompatibility and biodegradability, (iv) sustained controlled release, and (v) multidrug-loaded system. Stable nanocolloids of Paclitaxel (PTX) and lapatinib were prepared by the SLBL method. In a multidrug-resistant (MDR) ovarian cancer cell line, OVCAR-3, lapatinib/PTX nanocolloids mediated an enhanced cell growth inhibition in comparison with the PTX-only treatment. A series of in vitro cell assays were used to test the efficacy of these formulations. The small size and functional versatility of these nanoparticles, combined with their ability to incorporate various drugs, indicates that lapatinib/PTX nanocolloids may have in vivo therapeutic applications

Cytomechanical and topological investigation of MCF-7 cells by scanning force microscopy.

Despite enormous advances in breast cancer biology, there is an increased demand for new technologies/methods that are able to provide supplementary information to genomics and proteomics. Here, we exploit scanning force microscopy (SFM) in combination with confocal microscopy, to investigate the morphological and mechanical properties of two neoplastic cell lines: (i) MCF-7 (human breast cancer) and (ii) HeLa (human cervical carcinoma). Living and fixed cells either in phosphate buffer solution (PBS) or in air have been studied, and the viscoelastic properties (including the Young's modulus) of cells grown onto standard and modified (e.g. by fibronectin, one of the cellular matrix components) substrates have been measured. We observed different Young's modulus values, influenced by the adhesion and growth behaviour onto specific substrate surfaces

Properties of Nanocrystals-formulated Aluminosilicate Bricks

In the present work, seven different types of nanocrystals were studied as additives in the formulation of aluminosilicate bricks. The considered nanocrystals consisted of anatase titanium dioxide (two differently shaped types), boron modified anatase, calcium carbonate (in calcite phase), aluminium hydroxide and silicon carbide (of two diverse sizes), which were prepared using different methods. Syntheses aim to give a good control over a particle’s size and shape. Anatase titania nanocrystals, together with the nano-aluminium hydroxide ones, were synthesized via microwave-assisted procedures, with the use of different additives and without the final calcination steps. The silicon carbide nanoparticles were prepared via laser pyrolysis. The nano-calcium carbonate was prepared via a spray drying technique. All of the nanocrystals were tested as fillers (in 0.5, 1 and 2 wt. % amounts) in a commercial aluminosilicate refractory (55 % Al2O3, 42 % SiO2). They were used to prepare bricks that were thermally treated at 1300 °C for 24 hours, according to the international norms. The differently synthesized nanocrystals were added for the preparation of the bricks, with the aim to improve their heat-insulating and/or mechanical properties. The nanocrystals-modified refractories showed variations in properties, with respect to the untreated aluminosilicate reference in heat-insulating performances (thermal diffusivities were measured by the “hot disk” technique). In general, they also showed improvements in mechanical compression resistance for all of the samples at 2 wt. %. The best heat insulation was obtained with the addition of nano-aluminium hydroxide at 2 wt. %, while the highest mechanical compression breaking resistance was found with nano-CaCO3 at 2 wt. %. These outcomes were investigated with complementary techniques, like mercury porosimetry for porosity, and Archimedes methods to measure physical properties like the bulk and apparent densities, apparent porosities and water absorption. The results show that the nano-aluminium hydroxide modified bricks were the most porous, which could explain the best heat-insulating performances. There is a less straightforward explanation for the mechanical resistance results, as they may have relations with the characteristics of the pores. Furthermore, the nanoparticles may have possible reactions with the matrix during the heat treatments

Halloysite clay nanotubes as carriers for resveratrol delivery to tumoral cells

none3Halloysite is natural aluminosilicate clay with hollow tubular structure which allows loading with low soluble drugs using their saturated solutions in organic solvents. Resveratrol, a polyphenol known for having antioxidant and antineoplastic properties, is loaded inside these clay nanotubes lumens. Release time of 48 h is demonstrated. Spectroscopic and ζ-potential measurements are used to study the drug loading/release and for monitoring the nanotube layer-by-layer (LbL) coating with polyelectrolytes for further release control. Resveratrol-loaded clay nanotubes are added to breast cell cultures for toxicity tests. Halloysite functionalization with LbL polyelectrolyte multilayers remarkably decrease nanotube self-toxicity. MTT measurements performed with a neoplastic cell lines model system (MCF-7) as function of the resveratrol-loaded nanotubes concentration and incubation time indicate that drug-loaded halloysite strongly increase of cytotoxicity leading to cell apoptosis.Vergaro, Viviana; Lvov, Yuri M; Leporatti, StefanoVergaro, Viviana; Lvov, Yuri M; Leporatti, Stefan

Controlled biocide release from smart delivery systems: materials engineering to tune release rate, biointeractions, and responsiveness

Over the past 70 years, drug delivery systems (DDSs) have contributed to the development of precision medicine by delivering therapeutic molecules in a controlled manner. The literature reports several systems such as micelles, exosomes, liposomes, nanogels, metallic nanoparticles (NPs), polymers and copolymers, microemulsions, and so on, which are useful for medical and pharmaceutical purposes. These materials are used for targeted drug delivery to tissues and organs; for the sensitive detection of key biological molecules; for the more precise and safer imaging of diseased tissues; and for developing novel therapeutics for treating cancer, diabetes, pain, asthma, allergy, infection, and more. Many of these smart systems have aroused great interest in the agricultural sector. Derived from research focusing on the development of DDSs in the biomedical field, controlled release formulations (CRFs) of biocides have been used in the protection of agricultural crops and wood materials. Biocide is a general term describing an active substance used as a disinfectant, antiseptic, or preservative that has an important role in managing pathogens. These substances can deter, render harmless,inhibit the growth of, or kill harmful and unwanted microorganisms such as molds, bacteria, algae, insects, and rodents. Therefore, biocides are used in everyday life to protect, clean, and disinfect and are useful for different applications in a variety of industrials fields such as health care, food, pharmaceutical industries, and agriculture