ME T H O D S I N MO L E C U L A R B I O L O G Y

Series EditorJohn M. Walker

School of Life and Medical SciencesUniversity of HertfordshireHatfield, Hertfordshire, UK

For further volumes:http://www.springer.com/series/7651

For over 35 years, biological scientists have come to rely on the research protocols andmethodologies in the critically acclaimedMethods in Molecular Biology series. The series wasthe first to introduce the step-by-step protocols approach that has become the standard in allbiomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagentsneeded to complete the experiment, and followed by a detailed procedure that is supportedwith a helpful notes section offering tips and tricks of the trade as well as troubleshootingadvice. These hallmark features were introduced by series editor Dr. John Walker andconstitute the key ingredient in each and every volume of the Methods in Molecular Biologyseries. Tested and trusted, comprehensive and reliable, all protocols from the series areindexed in PubMed.

Nanoparticles in Biologyand Medicine

Methods and Protocols

Second Edition

Edited by

Enrico Ferrari

College of Science, School of Life Sciences, University of Lincoln, Brayford Pool, Lincoln, Lincolnshire, UK

Mikhail Soloviev

Centre for Biomedical Sciences, Department of Biological Sciences, Royal Holloway University of London,Egham, Surrey, UK

EditorsEnrico FerrariCollege of ScienceSchool of Life SciencesUniversity of LincolnBrayford PoolLincoln, Lincolnshire, UK

Mikhail SolovievCentre for Biomedical SciencesDepartment of Biological SciencesRoyal Holloway University of LondonEgham, Surrey, UK

ISSN 1064-3745 ISSN 1940-6029 (electronic)Methods in Molecular BiologyISBN 978-1-0716-0318-5 ISBN 978-1-0716-0319-2 (eBook)https://doi.org/10.1007/978-1-0716-0319-2

© Springer Science+Business Media, LLC, part of Springer Nature 2020This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material isconcerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproductionon microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation,computer software, or by similar or dissimilar methodology now known or hereafter developed.The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply,even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulationsand therefore free for general use.The publisher, the authors, and the editors are safe to assume that the advice and information in this book are believed tobe true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty,expressed or implied, with respect to the material contained herein or for any errors or omissions that may have beenmade. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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Preface

The modern fascination with micro- and nano-sized materials can be traced back to the1960s and 1970s when the first few attempts to use nanoparticles for controlled drugdelivery were reported. Since then, the use of nanoparticles in biology and medicine hasflourished, with a number of drug-containing nanoparticle products eventually reaching themarket in the past decade. Since the early 1990s, the number of publications on nanopar-ticles, their properties, and their applications in chemistry, material sciences, physics, medi-cine, and life sciences has grown exponentially. As there is no sign of slowing down, thisupdated second edition to the volume first published in 2012 is essential.

The term “nanoparticles” refers to a very special intermediate category between atomsor molecules and bulk material. While the term could be applied to any particle with adimension of less than 1 μm, it is more commonly used to describe particles ranging fromapproximately 1–100 nm. The uniquely small size of nanoparticles, which is of the sameorder as the wavelength of the electron wave function, results in quantum confinementeffects and consequently in the unique chemical, physical, and optical properties of nanos-caled materials. These are especially prominent for smaller clusters made of between ~100and ~200 atoms and having dimensions of about 3 nm, referred to as nanoclusters, wherequantum confinement effects are particularly prominent. Physical properties of nanoparti-cles and nanoclusters differ substantially from both bulk and molecular properties of thesame material. As the surface-to-volume ratio of a particle increases with the shrinking ofparticle size, an increasing number of individual atoms become exposed on the surface of aparticle compared to the number of atoms or molecules remaining buried inside it.

The range of materials explored for their unique nanoparticles properties is vast andincludes noble metals and semiconductors, inorganic and organic compounds, polymersand biopolymers, lipids and surfactants, proteins and peptides, to name just a few. The hugerange of chemical properties, further expanded through the nanoscale dimensions and theadditional options related to derivatization of nanoparticles for life science applications,brings about an immense array of opportunities which continue to grow. In biologylaboratories, nanoparticles are now routine reagents applied to imaging (quantum dots),transfection, and in vitro labeling of biomolecules or cells. They are also key components ofmany diagnostic kits, for example those based on lateral flow assays, which exploit theunique optical properties of gold nanoparticles. In vivo, nanoparticles represent anextremely promising tool for medical imaging (magnetic nanoparticles and nano-bubbles)and drug delivery. Nanoparticles can significantly improve the therapeutic value of impor-tant drugs, such as those used in cancer therapy. Nano-carriers have proven successful inseveral contexts because they can adsorb or encapsulate large amounts of those drugs thatwould otherwise have poor pharmacokinetics or too elevated toxicity if used on their own.Due to the nano-carriers small size, the embedded drug can still diffuse well in tissues,undergo endocytosis, and reach the target effectively. Also, due to the many nanoparticlechemistries available, it is increasingly possible to modulate the desired biodistribution of thepayload drug, derivatize the carrier with antibodies to promote accumulation in targettissues, and even combine the therapeutic effect of the drug with medical diagnostic ability(theragnostic).

v

This volume presents a wide range of methods for synthesis, surface modification,characterization, and application of nano-sized materials (nanoparticles) in life science andmedical fields (mostly for drug delivery and diagnostics). The chapters are organized in threeparts: (Part I) Synthesis of nanoparticles and their applications in biology and medicine(Chapters 1–13); (Part II) Focus on nanoparticle derivatization, bio-interface, and nano-toxicity (Chapters 14–20), and (Part III) Nanoparticle characterization and advancedmethods development (Chapters 21–29).

While maintaining the chapters of the previous edition that are still current and ofgeneral interest to life scientists, biomedical professionals, material scientists, and those withphysics and chemistry backgrounds, the new edition includes updates relevant to fastevolving areas of study. The second edition also includes background material that helpsin building a fundamental understanding of the nanoscale world (Chapters 1, 26, and 27).The book contains typical and detailed examples of methods suitable for use with gold(Chapters 1, 5, 14–17, 19–21, 23, and 26), silver (Chapters 2, 17, 19, 24, and 28–29),titanium oxide (Chapters 28 and 29), semiconductors (Chapter 25), silica-based nanoma-terials (Chapters 4, 5, and 9), methacrylate-based nanomaterials (Chapters 12 and 13),peptide and protein-based nanoparticles (Chapters 3 and 5–6), as well as a range of ferrousand magnetic materials (Chapters 13, 18, 20, and 27). Analytical science practitioners willhave a field day reading about the use of traditional UV-Vis-NIR, FTIR, and fluorescencespectrometry techniques (Chapters 2, 5–7, 9, 11, 15–16, 19, 24–26, and 29), light scatter-ing, Zeta potential, and other particle analyzers (Chapters 4–6, 8–9, 11–12, 15, 24, 26, and28–29), and TEM, STEM, and their variations (Chapters 5–6, 9–12, 24, and 26). Theapplications of particle tracking analysis are described in Chapter 22, and the ingeniouslyinventive use of traditional bright and dark field microscopy and flow cytometry for trackingnanoparticles is described in Chapters 28 and 29. A wide range of traditional biochemicalassays (Chapters 3, 5, 8, 10–12, and 17), immunoaffinity methods (Chapters 9, 12, and 18),and in vitro (Chapters 2–8, 10–12, 16, 18–19, and 26) and in vivo applications (Chapters 5,9, and 20) are also described. High precision quantification, combined with completechemical analyses, is described in Chapter 21; a range of nanoscaled electrochemical assaysand nano(mechanical) impact studies can be found in Chapters 23 and 24. Directed andhierarchical immobilization methods, designed to overcome limitations of traditionaluncontrolled processes of absorption of biopolymers, such as proteins onto gold nanopar-ticles, are now available (Chapters 16 and 17); these are designed to fully preserve theproven structure and function of the immobilized proteins. Their functional and structuralanalyses are described in Chapters 15, 16, and 25. Nanomaterials provide useful and uniquetools for the delivery of bioactive molecules such as nucleic acids, for example with hya-luronic acid nanoparticles for transfection of MicroRNA (Chapter 7), or the synthesis oflipid:peptide:DNA nanoparticles (Chapter 6). One additional focus in this edition is on thestudy of the bio-interface that determines the specific and complex interactions betweennanoparticles and living organisms. The book describes a range of typical and advancedmethods for the derivatization of nanoparticles with active biomolecules (Chapters 15–17)and the applications of molecule-nanoparticle interaction assays for diagnostic purposes orfor the determination of nanotoxicity effects (Chapters 18–20). For the first time, we alsoconsidered computational modeling and simulation of nanoparticle-biomolecule interac-tions, along with the other wet-lab methods, and included a step-by-step practical guideaimed at those who want to learn how to use molecular dynamics simulations (Chapter 14).

All methods chapters are written and presented with the view to help users to learn thesetechniques, to be able to repeat, and, if necessary, to modify the key technologies and adapt

vi Preface

them to particular needs. This volume will help beginners become familiar with thisfascinating field of research and will provide scientists at all levels of expertise with easy-to-follow practical advice needed tomake, modify, and analyze nanoparticles of their choice andto use them in a wide range of biomedical and pharmaceutical applications includingfunctional protein studies, drug delivery, immunochemistry, imaging, and many others.

We would like to express our gratitude to all the authors who have contributedinvaluably to this volume.

Lincoln, UK Enrico FerrariEgham, UK Mikhail Soloviev

Preface vii

Contents

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vContributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiii

PART I SYNTHESIS OF NANOPARTICLES AND THEIR APPLICATIONS

IN BIOLOGY AND MEDICINE

1 Procedures for the Synthesis and Capping of Metal Nanoparticles . . . . . . . . . . . . 3Claudia Gutierrez-Wing, J. Jesus Velazquez-Salazar,and Miguel Jose-Yacaman

2 Antimicrobial Applications of Silver Nanoparticlesto E. coli Colony Biofilms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21James P. McEvoy, Kayra Genc, Priya Loi, and William J. Walker

3 Elastins-Based Antimicrobial Particles for Deliveryof Bioactive Compounds. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29Raul Machado, Andre da Costa, Ana Margarida Pereira,Jose Carlos Rodriguez-Cabello, and Margarida Casal

4 Biomimetic Lipid Polymer Nanoparticles for Drug Delivery. . . . . . . . . . . . . . . . . . 45Ana Maria Carmona-Ribeiro

5 Crotamine Cell-Penetrating Nanocarriers: Cancer-Targetingand Potential Biotechnological and/or Medical Applications . . . . . . . . . . . . . . . . . 61Mirian A. F. Hayashi, Joana Darc Campeiro, Lucas Carvalho Porta,Brian Szychowski, Wendel Andrade Alves, Eduardo B. Oliveira,Irina Kerkis, Marie-Christine Daniel, and Richard L. Karpel

6 Preparation of Lipid–Peptide–DNA (LPD) Nanoparticlesand Their Use for Gene Transfection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91Fan Zhang and Hao-Ying Li

7 Preparation of Hyaluronic Acid-Based Nanoparticlesfor Macrophage-Targeted MicroRNA Delivery and Transfection. . . . . . . . . . . . . . 99Neha N. Parayath and Mansoor M. Amiji

8 Formulation and Characterization of AntithrombinPerfluorocarbon Nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111Alexander J. Wilson, Qingyu Zhou, Ian Vargas, Rohun Palekar,Ryan Grabau, Hua Pan, and Samuel A. Wickline

9 Biomedical In Vivo Studies with ORMOSIL NanoparticlesContaining Active Agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121Sona Gandhi and Indrajit Roy

10 Preparation of Spray-Dried Nanoparticles for EfficientDrug Delivery to the Lungs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139Hao-Ying Li and Fan Zhang

ix

11 Synthesis and Evaluation of Airway-Targeted PLGA-PEGNanoparticles for Drug Delivery in Obstructive Lung Diseases . . . . . . . . . . . . . . . 147Neeraj Vij

12 Synthesis and Evaluation of Dendrimers for AutophagyAugmentation and Alleviation of Obstructive Lung Diseases . . . . . . . . . . . . . . . . . 155Neeraj Vij

13 Synthesis of Poly(2-Hydroxyethyl Methacrylate) (PHEMA)-BasedSuperparamagnetic Nanoparticles for Biomedical and PharmaceuticalApplications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165Rajesh K. Saini, Jaya Bajpai, and Anil K. Bajpai

PART II FOCUS ON THE NANOPARTICLES DERIVATIZATION,BIO-INTERFACE, AND NANOTOXICITY

14 The Molecular Dynamics Simulation of Peptideson Gold Nanosurfaces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177Danilo Roccatano

15 Protein Immobilization on Gold Nanoparticles:Quantitative Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199Evan Decker, Chunsheng Bai, Lauren Nelless, Enrico Ferrari,and Mikhail Soloviev

16 Oriented Immobilization on Gold Nanoparticlesof a Recombinant Therapeutic Zymogen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213Elina Dosadina, Celetia Agyeiwaa, William Ferreira, Simon Cutting,Abdullah Jibawi, Enrico Ferrari, and Mikhail Soloviev

17 Directed and Modular Protein Immobilization on Goldand Silver Nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227Angela Saccardo, Wenwei Ma, Mikhail Soloviev, and Enrico Ferrari

18 In Vitro Labeling Mesenchymal Stem Cells with SuperparamagneticIron Oxide Nanoparticles: Efficacy and Cytotoxicity . . . . . . . . . . . . . . . . . . . . . . . . 235Jasmin

19 RNA Quantification Using Noble Metal Nanoprobes: SimultaneousIdentification of Several Different mRNA Targets UsingColor Multiplexing and Application to Chronic MyeloidLeukemia Diagnostics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251Pedro Viana Baptista

20 Assessment of Toxicity of Nanoparticles Using Insectsas Biological Models . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 269Yan Zhou, Yan Chen, Aracely Rocha, Carlos J. Sanchez, and Hong Liang

PART III NANOPARTICLES CHARACTERIZATION AND ADVANCED

METHODS DEVELOPMENT

21 Absolute Quantification of Gold Nanoparticles with FemtomolarAccuracy Using Inductively Coupled Plasma AtomicEmission Spectroscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 283Lee-Anne McCarthy, Andrew Dye, and Enrico Ferrari

x Contents

22 Nanoparticle Tracking Analysis for Multiparameter Characterizationand Counting of Nanoparticle Suspensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 289Duncan Griffiths, Pauline Carnell-Morris, and Matthew Wright

23 Nanoparticle Bridges for Studying Electrical Properties of OrganicMolecules and Gas Sensor Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305Klaus Leifer, Syed Hassan Mujtaba Jafri, and Yuanyuan Han

24 Nanoparticle Characterization Through Nano-ImpactElectrochemistry: Tools and Methodology Development . . . . . . . . . . . . . . . . . . . . 327Kevin A. Kirk, Tulashi Luitel, Farideh Hosseini Narouei,and Silvana Andreescu

25 Photostability of Semiconductor Quantum Dots in Responseto UV Exposure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 343Julian Bailes

26 Gold Nanoclusters, Gold Nanoparticles, and Analytical Techniquesfor Their Characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 351German Plascencia-Villa, Ruben Mendoza-Cruz, Lourdes Bazan-Dıaz,and Miguel Jose-Yacaman

27 Combining Nanoparticles with Colloidal Bubbles: A Short Review . . . . . . . . . . . 383Ekaterina Litau

28 Combination of Dark-Field and Confocal Microscopyfor the Optical Detection of Silver and Titanium Nanoparticlesin Mammalian Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395Robert Martin Zucker and William K. Boyes

29 Detection of Silver and TiO2 Nanoparticles in Cellsby Flow Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 415Robert Martin Zucker and William K. Boyes

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 437

Contents xi

Contributors

CELETIA AGYEIWAA • Centre for Biomedical Sciences, School of Biological Sciences, RoyalHolloway University of London, Egham, Surrey, UK

WENDEL ANDRADE ALVES • Centro de Ciencias Naturais e Humanas, Universidade Federaldo ABC, Santo Andre, SP, Brazil

MANSOOR M. AMIJI • Department of Pharmaceutical Sciences, School of Pharmacy,Northeastern University, Boston, MA, USA

SILVANA ANDREESCU • Department of Chemistry and Biomolecular Science, ClarksonUniversity, Potsdam, NY, USA

CHUNSHENG BAI • College of Science, University of Lincoln, Lincoln, UKJULIAN BAILES • Centre for Biomedical Sciences, School of Biological Sciences, Royal Holloway

University of London, Egham, Surrey, UKANIL K. BAJPAI • Bose Memorial Research Laboratory, Department of Chemistry,

Government Autonomous Science College, Jabalpur, MP, IndiaJAYA BAJPAI • Bose Memorial Research Laboratory, Department of Chemistry, Government

Autonomous Science College, Jabalpur, MP, IndiaPEDRO VIANA BAPTISTA • Departamento de Ciencias da Vida, Faculdade de Ciencias

e Tecnologia, Universidade Nova de Lisboa, UCIBIO, Caparica, PortugalLOURDES BAZAN-DIAZ • Instituto de Investigaciones en Materiales (IIM), Universidad

Nacional Aut�onoma de Mexico (UNAM), Mexico City, TX, USAWILLIAM K. BOYES • Neurological and Endocrine Toxicology Branch, Public Health and

Integrated Toxicology Division, Center for Public Health and Environmental Assessment,Office of Research and Development U.S. Environmental Protection Agency, ResearchTriangle Park, NC, USA

ANA MARIA CARMONA-RIBEIRO • Biocolloids Laboratory, Departamento de Bioquımica,Instituto de Quımica, Universidade de Sao Paulo, Sao Paulo, SP, Brazil

PAULINE CARNELL-MORRIS • Malvern Panalytical Ltd., Malvern, UKMARGARIDA CASAL • CBMA (Centre of Molecular and Environmental Biology), Department

of Biology, University of Minho, Braga, Portugal; IB-S (Institute of Science and Innovationfor Sustainability), University of Minho, Braga, Portugal

YAN CHEN • Materials Science andMechanical Engineering, Texas A&MUniversity, CollegeStation, TX, USA

SIMON CUTTING • Centre for Biomedical Sciences, School of Biological Sciences, RoyalHolloway University of London, Egham, Surrey, UK

ANDRE DA COSTA • CBMA (Centre of Molecular and Environmental Biology), Departmentof Biology, University of Minho, Braga, Portugal; IB-S (Institute of Science and Innovationfor Sustainability), University of Minho, Braga, Portugal

MARIE-CHRISTINE DANIEL • Department of Chemistry and Biochemistry, University ofMaryland Baltimore County, Baltimore, MD, USA

JOANA DARC CAMPEIRO • Departamento de Farmacologia, Universidade Federal de SaoPaulo (UNIFESP), Sao Paulo, SP, Brazil

EVAN DECKER • Centre for Biomedical Sciences, School of Biological Sciences, Royal HollowayUniversity of London, Egham, Surrey, UK

xiii

ELINA DOSADINA • Centre for Biomedical Sciences, School of Biological Sciences, RoyalHolloway University of London, Egham, Surrey, UK

ANDREW DYE • College of Science, University of Lincoln, Lincoln, UKENRICO FERRARI • College of Science, School of Life Sciences, University of Lincoln, Brayford

Pool, Lincoln, Lincolnshire, UKWILLIAM FERREIRA • Centre for Biomedical Sciences, School of Biological Sciences, Royal

Holloway University of London, Egham, Surrey, UKSONA GANDHI • Department of Chemistry, University of Delhi, Delhi, IndiaKAYRA GENC • Centre for Biomedical Sciences, School of Biological Sciences, Royal Holloway,

University of London, Egham, Surrey, UKRYAN GRABAU • The USF Health Heart Institute, University of South Florida, Tampa, FL,

USADUNCAN GRIFFITHS • Malvern Panalytical Ltd., Malvern, UKCLAUDIA GUTIERREZ-WING • Instituto Nacional de Investigaciones Nucleares, Ciencias

Aplicadas-Tecnologıa de Materiales, La Marquesa Ocoyoacac, Edo. de Mexico, MexicoYUANYUAN HAN • Division of Applied Materials Science, Department of Engineering

Sciences, Uppsala University, Uppsala, SwedenMIRIAN A. F. HAYASHI • Departamento de Farmacologia, Universidade Federal de Sao Paulo

(UNIFESP), Sao Paulo, SP, BrazilSYED HASSAN MUJTABA JAFRI • Division of Applied Materials Science, Department of

Engineering Sciences, Uppsala University, Uppsala, Sweden; Department of ElectricalEngineering, Mirpur University of Science and Technology, Mirpur Azad Jammu andKashmir, Pakistan

JASMIN • Nucleo Multidisciplinar de Pesquisa em Biologia, Universidade Federal do Rio deJaneiro, Duque de Caxias, Rio de Janeiro, Brazil

ABDULLAH JIBAWI • Ashford and St. Peter’s Hospitals NHS Foundation Trust, Surrey, UKMIGUEL JOSE-YACAMAN • Department of Physics and Astronomy, University of Texas at San

Antonio, San Antonio, TX, USA; Department of Applied Physics and Material Science,Northern Arizona University, Flagstaff, USA

RICHARD L. KARPEL • Department of Chemistry and Biochemistry, University of MarylandBaltimore County, Baltimore, MD, USA

IRINA KERKIS • Laboratory of Genetics, Butantan Institute, Sao Paulo, SP, BrazilKEVIN A. KIRK • Department of Chemistry and Biomolecular Science, Clarkson University,

Potsdam, NY, USAKLAUS LEIFER • Division of Applied Materials Science, Department of Engineering Sciences,

Uppsala University, Uppsala, SwedenHAO-YING LI • Biomanufacturing Research Centre, School of Mechanical and Electronic

Engineering, Soochow University, Suzhou, ChinaHONG LIANG • Materials Science and Mechanical Engineering, Texas A&M University,

College Station, TX, USAEKATERINA LITAU • Royal Holloway University of London, Egham, Surrey, UKPRIYA LOI • Centre for Biomedical Sciences, School of Biological Sciences, Royal Holloway,

University of London, Egham, Surrey, UKTULASHI LUITEL • Department of Chemistry and Biomolecular Science, Clarkson University,

Potsdam, NY, USAWENWEI MA • College of Science, University of Lincoln, Lincoln, UK

xiv Contributors

RAULMACHADO • CBMA (Centre of Molecular and Environmental Biology), Department ofBiology, University of Minho, Braga, Portugal; IB-S (Institute of Science and Innovationfor Sustainability), University of Minho, Braga, Portugal

LEE-ANNE MCCARTHY • College of Science, University of Lincoln, Lincoln, UKJAMES P. MCEVOY • Centre for Biomedical Sciences, School of Biological Sciences, Royal

Holloway, University of London, Egham, Surrey, UKRUBEN MENDOZA-CRUZ • Instituto de Investigaciones en Materiales (IIM), Universidad

Nacional Aut�onoma de Mexico (UNAM), Mexico City, TX, USAFARIDEH HOSSEINI NAROUEI • Department of Chemistry and Biomolecular Science, Clarkson

University, Potsdam, NY, USALAUREN NELLESS • Centre for Biomedical Sciences, School of Biological Sciences, Royal

Holloway University of London, Egham, Surrey, UKEDUARDO B. OLIVEIRA • Departamento de Bioquımica e Imunologia, Universidade de Sao

Paulo, Ribeirao Preto, SP, BrazilROHUN PALEKAR • Department of Biomedical Engineering, Washington University in St

Louis, St. Louis, MO, USAHUA PAN • The USF Health Heart Institute, University of South Florida, Tampa, FL, USANEHA N. PARAYATH • Department of Pharmaceutical Sciences, School of Pharmacy,

Northeastern University, Boston, MA, USA; Clinical Research Division, Fred HutchinsonCancer Research Center, Seattle, WA, USA

ANA MARGARIDA PEREIRA • CBMA (Centre of Molecular and Environmental Biology),Department of Biology, University of Minho, Braga, Portugal; IB-S (Institute of Scienceand Innovation for Sustainability), University of Minho, Braga, Portugal

GERMAN PLASCENCIA-VILLA • Department of Biology, The University of Texas at San Antonio(UTSA), San Antonio, TX, USA

LUCAS CARVALHO PORTA • Departamento de Farmacologia, Universidade Federal de SaoPaulo (UNIFESP), Sao Paulo, SP, Brazil

DANILO ROCCATANO • School of Mathematics and Physics, University of Lincoln, Lincoln, UKARACELY ROCHA • Materials Science and Mechanical Engineering, Texas A&M University,

College Station, TX, USAJOSE CARLOS RODRIGUEZ-CABELLO • Bioforge (Group for Advanced Materials and

Nanobiotechnology), Universidad de Valladolid, Valladolid, Spain; Networking ResearchCentre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Valladolid,Spain

INDRAJIT ROY • Department of Chemistry, University of Delhi, Delhi, IndiaANGELA SACCARDO • College of Science, University of Lincoln, Lincoln, UKRAJESH K. SAINI • Bose Memorial Research Laboratory, Department of Chemistry,

Government Autonomous Science College, Jabalpur, MP, IndiaCARLOS J. SANCHEZ • Materials Science and Mechanical Engineering, Texas A&M

University, College Station, TX, USAMIKHAIL SOLOVIEV • Centre for Biomedical Sciences, Department of Biological Sciences,

Royal Holloway University of London, Egham, Surrey, UKBRIAN SZYCHOWSKI • Department of Chemistry and Biochemistry, University of Maryland

Baltimore County, Baltimore, MD, USAIAN VARGAS • The USFHealth Heart Institute, University of South Florida, Tampa, FL, USAJ. JESUS VELAZQUEZ-SALAZAR • Department of Physics and Astronomy, University of Texas at

San Antonio, San Antonio, TX, USA

Contributors xv

NEERAJ VIJ • Department of Pediatrics and Pulmonary Medicine, The Johns HopkinsUniversity School of Medicine, Baltimore, MD, USA; 4Dx Limited, Los Angeles, CA, USA;VIJ BIOTECH & PRECISION THERANOSTICS INC, Baltimore, MD, USA

WILLIAM J. WALKER • Centre for Biomedical Sciences, School of Biological Sciences, RoyalHolloway, University of London, Egham, Surrey, UK

SAMUEL A.WICKLINE • The USFHealth Heart Institute, University of South Florida, Tampa,FL, USA

ALEXANDER J. WILSON • The USF Health Heart Institute, University of South Florida,Tampa, FL, USA

MATTHEW WRIGHT • Malvern Panalytical Ltd., Malvern, UKFAN ZHANG • Laboratory of Molecular Biology, Medical Research Council, Cambridge, UKQINGYU ZHOU • College of Pharmacy, University of South Florida, Tampa, FL, USAYAN ZHOU • Materials Science and Mechanical Engineering, Texas A&M University,

College Station, TX, USAROBERT MARTIN ZUCKER • Reproductive and Developmental Toxicology Branch,

Public Health and Integrated Toxicology Division, Center for Public Healthand Environmental Assessment, Office of Research and Development U.S. EnvironmentalProtection Agency, Research Triangle Park, NC, USA

xvi Contributors