The shielding effect of phospholipidic bilayers on zinc oxide nanocrystals for biomedical applications

Zinc oxide nanocrystals (ZnO NCs), thanks to their unique properties, are receiving much attention for their use in nanomedicine, in particular for therapy against cancer [1]. To be efficiently employed as diagnostic and therapeutic (yet theranostic) tools [2], highly dispersed, stable and non-toxic nanoparticles are required. In the case of ZnO NCs, there is still a lack of knowledge about cytotoxicity mechanisms and stability in the biological context, as well as immunological response and haemocompatible features. Most of these above-mentioned behaviours strongly depends on physico-chemical and surface properties of the nanoparticles. We thus propose a novel approach to stabilize the ZnO NCs in various biological media, focusing on NC aggregation and biodegradation as a function of the surface functionalization. We synthesized bare ZnO NCs, amino-propyl functionalized ones, and lipid bilayer-shielded NCs, and we characterized their morphological, chemical and physical properties. The stability behavior of the three different samples was evaluated, comparing their biodegradation profiles in different media, i.e. organic solvents, water, and different simulated and biological fluids. The studies aim to investigate how the particle surface functionalizations, and thus chemistry and charge, could influence their hydrodynamic size, zeta potential and consequent aggregation and degradation in the different solvents. We demonstrated that bare and amino-functionalized ZnO NCs strongly and rapidly aggregate when suspended in both simulated and biological media. Long-term biodegradation analysis showed small dissolution into potentially cytotoxic Zn-cations, also slightly affecting their crystalline structure. In contrast, high colloidal stability and integrity was retained for lipid-shielded ZnO NCs in all media, rendering them the ideal candidates for further theranostic applications [3]. [1] P. Zhu, Z. Weng, X. Li, X. Liu, S. Wu Adv. Mater. Interfaces 3 (2016) 1500494. [2] E. Lim, T. Kim, S. Paik, S. Haam, Y. Huh, and K. Lee, Chem. Rev. 115 (2015) 327−394. [3] B. Dumontel, M. Canta, H. Engelke, A. Chiodoni, L. Racca, A. Ancona, T. Limongi, G. Canavese and V. Cauda, J. Mater. Chem. B, under review The support from ERC Starting Grant – Project N. 678151 “Trojananohorse” and Compagnia di Sanpaolo are gratefully acknowledged

Lipid-coated zinc oxide nanocrystals as innovative ROS-generators for photodynamic therapy

Photodynamic Therapy (PDT) is a medical treatment that combines the administration of a nontoxic drug, called photosensitizer (PS), with light irradiation of the targeted region. It has been proposed as a new cancer therapy, promising better selectivity and fewer side-effects compared to traditional chemo- and radio-therapies. PSs indeed can accumulate specifically within the region of interest so that when the light is directly focused only in that region the therapeutic effect is highly localized. Traditional PSs, like chlorins and porphyrins, suffer from several drawbacks such as aggregation in biological media and poor biocompatibility. Thus, the development of innovative photosensitizers able to overcome these issues is crucial to the therapeutic action of PDT. Among the others, nanostructured Zinc Oxide (ZnO) has been recently proposed as new therapeutic agent and PS thanks to its semiconducting properties, biocompatible features, and ease of functionalization [1]. Nevertheless, further efforts are needed in order to improve its colloidal stability in biological media and to unravel the effective therapeutic mechanism. Here, we propose the synthesis and characterization of lipid-coated ZnO nanoparticles as new photosensitizer for cancer PDT [2]. First, by Dynamic Light Scattering (DLS) experiments, we show that the lipid-coating increases the colloidal stability of the ZnO NPs in Phosphate buffered saline (PBS). Then, using Electron Paramagnetic Resonance (EPR) coupled with the spin-trapping technique, we demonstrate and characterize the ability of bare and lipid-coated ZnO NPs to generate Reactive Oxygen Species (ROS) in water only when remotely actuated via light irradiation. Interestingly, our results aware that the surface chemistry of the NPs greatly influence the type of photo-generated ROS. Finally, we show that our NPs are effectively internalized inside human epithelial carcinoma cells (HeLa) via a lysosomal pathway and that they are able to generate ROS inside cancer cells. [1] B. Dumontel, M. Canta, H. Engelke, A. Chiodoni, L. Racca, A. Ancona, T. Limongi, G. Canavese and V. Cauda, ‎J. Mater. Chem. B. under revision. [2] A. Ancona, H. Engelke, N. Garino, B. Dumontel, W.Fazzini and V. Cauda, to be submitted. The support from ERC Starting Grant – Project N. 678151 “Trojananohorse” is gratefully acknowledged

Biomimetic Non-Immunogenic Nanoassembly for the Antitumor Therapy

Nanoassembly (1) for inducing apoptosis in cancer cells comprising: a core (2) comprising at least a nanoparticle of a nano structured and semiconductor metal oxide, said nanoparticle being monocrystalline or polycrystalline; a shell (3) formed by a double phospholipid layer and proteins derived from an extracellular biovesicole chosen between an exosome, an ectosome, a connectosome, an oncosome and an apoptotic body, and an oncosome, said core (2) being enclosed inside said shell (3); and a plurality of targeting molecules (4, 4', 4") of said cancer cells, preferably monoclonal antibodies (4, 4', 4"), said molecules (4, 4', 4") being anchored to the external surface of said biovesicole

Enhanced Biostability and Cellular Uptake of Zinc Oxide Nanocrystals Shielded with Phospholipid Bilayer

The widespread use of ZnO nanomaterials for biomedical applications, including therapeutic drug delivery or stimuli-responsive activation, as well as imaging, imposes a careful control over the colloidal stability and long-term behaviour of ZnO in biological media. Moreover, the effect of ZnO nanostructures on living cells, in particular cancer cells, is still under debate. This paper discusses the role of surface chemistry and charge of zinc oxide nanocrystals, of around 15 nm in size, which influence their behaviour in biological fluids and effect on cancer cells. In particular, we address this problem by modifying the surface of pristine ZnO nanocrystals (NCs), rich of hydroxyl groups, with positively charged amino-propyl chains or, more innovatively, by self-assembling a double-lipidic membrane, shielding the ZnO NCs. Our findings show that the prolonged immersion in simulated human plasma and in the cell culture medium leads to highly colloidally dispersed ZnO NCs only when coated by the lipidic bilayer. In contrast, the pristine and amine-functionalized NCs form huge aggregates after already one hour of immersion. Partial dissolution of these two samples into potentially cytotoxic Zn2+ cations takes place, together with the precipitation of phosphate and carbonate salts on the NCs’ surface. When exposed to living HeLa cancer cells, higher amounts of lipid-shielded ZnO NCs are internalized with respect to the other samples, thus showing a reduced cytotoxicity, based on the same amount of internalized NCs. These results pave the way for the development of novel theranostic platforms based on ZnO NCs. The new formulation of ZnO shielded with a lipid-bilayer will prevent strong aggregation and premature degradation into toxic by-products, and promote a highly efficient cell uptake for further therapeutic or diagnostic functions

Engineered extracellular vesicles as a reliable tool in cancer nanomedicine

Fast diagnosis and more ecient therapies for cancer surely represent one of the huge tasks for the worldwide researchers’ and clinicians’ community. In the last two decades, our understanding of the biology and molecular pathology of cancer mechanisms, coupled with the continuous development of the material science and technological compounds, have successfully improved nanomedicine applications in oncology. This review argues on nanomedicine application of engineered extracellular vesicles (EVs) in oncology. All the most innovative processes of EVs engineering are discussed together with the related degree of applicability for each one of them in cancer nanomedicines