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      Nanotoxicology and Nanosafety: Safety-by-Design and Testing at a Glance


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          This review offers a systematic discussion about nanotoxicology and nanosafety associated with nanomaterials during manufacture and further biomedical applications. A detailed introduction on nanomaterials and their most frequently uses, followed by the critical risk aspects related to regulatory uses and commercialization, is provided. Moreover, the impact of nanotoxicology in research over the last decades is discussed, together with the currently available toxicological methods in cell cultures (in vitro) and in living organisms (in vivo). A special focus is given to inorganic nanoparticles such as titanium dioxide nanoparticles (TiO 2NPs) and silver nanoparticles (AgNPs). In vitro and in vivo case studies for the selected nanoparticles are discussed. The final part of this work describes the significance of nano-security for both risk assessment and environmental nanosafety. “Safety-by-Design” is defined as a starting point consisting on the implementation of the principles of drug discovery and development. The concept “Safety-by-Design” appears to be a way to “ensure safety”, but the superficiality and the lack of articulation with which it is treated still raises many doubts. Although the approach of “Safety-by-Design” to the principles of drug development has helped in the assessment of the toxicity of nanomaterials, a combination of scientific efforts is constantly urgent to ensure the consistency of methods and processes. This will ensure that the quality of nanomaterials is controlled and their safe development is promoted. Safety issues are considered strategies for discovering novel toxicological-related mechanisms still needed to be promoted.

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          Cellular uptake of nanoparticles: journey inside the cell

          Cellular association and trafficking of nanoscale materials enables us to both understand and exploit context-dependent phenomena in various disease states, their pathogenesis, and potential therapeutic approaches. Nanoscale materials are increasingly found in consumer goods, electronics, and pharmaceuticals. While these particles interact with the body in myriad ways, their beneficial and/or deleterious effects ultimately arise from interactions at the cellular and subcellular level. Nanoparticles (NPs) can modulate cell fate, induce or prevent mutations, initiate cell–cell communication, and modulate cell structure in a manner dictated largely by phenomena at the nano–bio interface. Recent advances in chemical synthesis have yielded new nanoscale materials with precisely defined biochemical features, and emerging analytical techniques have shed light on nuanced and context-dependent nano-bio interactions within cells. In this review, we provide an objective and comprehensive account of our current understanding of the cellular uptake of NPs and the underlying parameters controlling the nano-cellular interactions, along with the available analytical techniques to follow and track these processes.
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            Characterization techniques for nanoparticles: comparison and complementarity upon studying nanoparticle properties

            Combined and carefully selected use of experimental techniques – understanding nanoparticle properties and optimizing performance in applications. Nanostructures have attracted huge interest as a rapidly growing class of materials for many applications. Several techniques have been used to characterize the size, crystal structure, elemental composition and a variety of other physical properties of nanoparticles. In several cases, there are physical properties that can be evaluated by more than one technique. Different strengths and limitations of each technique complicate the choice of the most suitable method, while often a combinatorial characterization approach is needed. In addition, given that the significance of nanoparticles in basic research and applications is constantly increasing, it is necessary that researchers from separate fields overcome the challenges in the reproducible and reliable characterization of nanomaterials, after their synthesis and further process ( e.g. annealing) stages. The principal objective of this review is to summarize the present knowledge on the use, advances, advantages and weaknesses of a large number of experimental techniques that are available for the characterization of nanoparticles. Different characterization techniques are classified according to the concept/group of the technique used, the information they can provide, or the materials that they are destined for. We describe the main characteristics of the techniques and their operation principles and we give various examples of their use, presenting them in a comparative mode, when possible, in relation to the property studied in each case.
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              Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation.

              Adult somatic tissues have proven difficult to expand in vitro, largely because of the complexity of recreating appropriate environmental signals in culture. We have overcome this problem recently and developed culture conditions for adult stem cells that allow the long-term expansion of adult primary tissues from small intestine, stomach, liver and pancreas into self-assembling 3D structures that we have termed 'organoids'. We provide a detailed protocol that describes how to grow adult mouse and human liver and pancreas organoids, from cell isolation and long-term expansion to genetic manipulation in vitro. Liver and pancreas cells grow in a gel-based extracellular matrix (ECM) and a defined medium. The cells can self-organize into organoids that self-renew in vitro while retaining their tissue-of-origin commitment, genetic stability and potential to differentiate into functional cells in vitro (hepatocytes) and in vivo (hepatocytes and endocrine cells). Genetic modification of these organoids opens up avenues for the manipulation of adult stem cells in vitro, which could facilitate the study of human biology and allow gene correction for regenerative medicine purposes. The complete protocol takes 1-4 weeks to generate self-renewing 3D organoids and to perform genetic manipulation experiments. Personnel with basic scientific training can conduct this protocol.

                Author and article information

                Int J Environ Res Public Health
                Int J Environ Res Public Health
                International Journal of Environmental Research and Public Health
                28 June 2020
                July 2020
                : 17
                : 13
                : 4657
                [1 ]Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal; zielinska-aleksandra@ 123456wp.pl (A.Z.); beafecosta@ 123456gmail.com (B.C.); mariavferreira00@ 123456gmail.com (M.V.F.); diogodmigueis@ 123456gmail.com (D.M.); jessicamslouros@ 123456gmail.com (J.M.S.L.)
                [2 ]Institute of Human Genetics, Polish Academy of Sciences, Strzeszyńska 32, 60-479 Poznań, Poland
                [3 ]CREA-Research Centre for Food and Nutrition, Via Ardeatina 546, 00178 Rome, Italy; alessandra.durazzo@ 123456crea.gov.it (A.D.); massimo.lucarini@ 123456crea.gov.it (M.L.)
                [4 ]Department of Gastroenterology, Dietetics and Internal Diseases, Poznan University of Medical Sciences, Przybyszewskiego 49, 60-355 Poznań, Poland; piotr.eder@ 123456op.pl
                [5 ]Laboratory of Biomaterials and Nanotechnology, University of Sorocaba—UNISO, Sorocaba 18023-000, Brazil; marco.chaud@ 123456prof.uniso.br
                [6 ]Center for Biomedical Engineering, Department of Medicine, Brigham and Women& Hospital, Harvard Medical School, 65 Landsdowne Street, Cambridge, MA 02139, USA; m.a.j.morsink@ 123456student.utwente.nl (M.M.); n.g.a.willemen@ 123456student.utwente.nl (N.W.); pattypharma@ 123456gmail.com (P.S.)
                [7 ]Translational Liver Research, Department of Medical Cell BioPhysics, Technical Medical Centre, Faculty of Science and Technology, University of Twente, 7522 NB Enschede, The Netherlands
                [8 ]Department of Developmental BioEngineering, Faculty of Science and Technology, Technical Medical Centre, University of Twente, 7522 NB Enschede, The Netherlands
                [9 ]Nanomedicine and Nanotechnology Laboratory (LNMed), Institute of Technology and Research (ITP), University of Tiradentes (Unit), Av. Murilo Dantas, 300, Aracaju 49010-390, Brazil
                [10 ]Tiradentes Institute, 150 Mt Vernon St, Dorchester, MA 02125, USA
                [11 ]Department of Pharmacy, University of Napoli Federico II, 80131 Napoli, Italy
                [12 ]CEB—Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal
                Author notes
                [* ]Correspondence: asantini@ 123456unina.it (A.S.); ebsouto@ 123456ff.uc.pt (E.B.S.); Tel.: +39-81-253-9317 (A.S.); +351-239-488-400 (E.B.S.)
                Author information
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                : 09 May 2020
                : 23 June 2020

                Public health
                nanotoxicology,nanosafety,nanomaterials,risk assessment,toxicity tests,nanoparticles,human health,safety-by-design,biological systems


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