Antibacterial, plasmonic, and toxic properties of engineered nanoparticles

ANTIBACTERIAL, PLASMONIC, AND TOXIC PROPERTIES OF ENGINEERED NANOPARTICLES A Thesis presented to the Faculty of the Graduate School University of Missouri In Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy by Trang Ha Dieu Nguyen Drs Mengshi Lin, Azlin Mustapha Thesis Supervisors DECEMBER 2016 © Copyright by Trang Ha Dieu Nguyen 2016 All rights reserved The undersigned, appointed by the dean of the Graduate School, have examined the thesis entitled ANTIBACTERIAL, PLASMONIC AND TOXIC PROPERTIES OF ENGINEERED NANOPARTICLES Presented by Trang Ha Dieu Nguyen, a candidate for the degree of Doctor of Philosophy, and hereby certify that, in their opinion, it is worthy of acceptance Mengshi Lin, Ph.D., Food Science Program Azlin Mustapha, Ph.D., Food Science Program Bongkosh Vardhanabhuti, Ph.D., Food Science Program Chong He, Ph.D., Department of Statistics ACKNOWLEDGEMENTS In the heat of moment, when many ideas for the acknowledgment part of my dissertation come along, I was so excited This dissertation took me four and a half years and it has given me much of challenges and joyfulness This work would not have been possible without the guidance and supports of all the following individuals First and foremost, I would like to thank my two wonderful advisors, Drs Mengshi Lin and Azlin Mustapha Dr Lin is a facilitator and a mentor who abundantly helped and offered me invaluable assistance, supports and encouragements during my time in Mizzou I believe he spent hundreds of hours editing my manuscripts, even in the weekends Dr Mustapha inspired me by her enthusiasm, energy, and excellent knowledge I cannot fulfill my dissertation without her advice and guidance when I had problems with my experiments She is the person who has proven that a female scientist can balance well between her family and career! I would like to show my gratitude to my committee members, Dr Vardhanabhuti and Dr He through the journey I appreciate their valuable suggestions and comments, especially Dr V for her co-authorship in one of my manuscripts I also would like to express my thanks to Dr Koc, Dr Clarke, Dr Gruen, Dr Elmore, Dr Alexander for letting me help them in teaching assistance I have learned from them teaching methods, curricula and appropriate manner when interacting with students ii I thank my past and present lab members and from different labs for being wonderful colleagues to work with Special thanks are given to Zhang Zhong for his co-authorship and assistance I acknowledge the Vietnam International Education Development under the Ministry of Education and Training for financial support in the first two years of my study Finally, I owe my deepest gratitude to my family in Vietnam and my husband’s family in the US for endless love, support, and encouragement, without which I could not finish my work My husband and my sons Yanni and Raphael are the greatest gifts that God brought to me Thank you all for being with me, loving me unconditionally, being my support and inspiring me every single day of my life iii TABLE OF CONTENTS ACKNOWLEDGEMENTS ii LIST OF TABLES vii LIST OF FIGURES viii ABSTRACT xi CHAPTER Introduction 1.1 Background 1.2 Objectives CHAPTER Literature review 2.1 Nanomaterials and current uses in foods and consumer products 2.2 Surface enhanced Raman spectroscopy (SERS) and its enhancement mechanisms 13 2.3 Applications of SERS for Food Adulterant Detection 17 2.4 SERS substrates 19 2.5 Antibacterial properties of inorganic ENPs 20 2.6 Mechanisms of antimicrobial properties of inorganic ENPs 26 2.7 Physical and chemical properties of inorganic ENPS affect their antimicrobial activities 29 2.8 Cytotoxicity of inorganic NPs to human cells 32 2.9 Mechanisms of cytoxocity of ENPS 35 2.10 Physical and chemical properties of NMs associated with nanotoxicity 37 2.11 Toxicity Assessment of NPs 40 2.11.1 Cell uptake 40 2.11.2 Cell viability 41 2.11.3 Cell functions 42 CHAPTER 49 Use of Graphene and Gold Nanorods as Substrates for Detection of Pesticides by Surface Enhanced Raman Spectroscopy 49 iv 3.1 Introduction 51 3.2 Materials and Methods 53 3.2.1 Preparation of chemicals 53 3.2.2 Gold film silicon substrate 54 3.2.3 Synthesis of gold nanorods 54 3.2.4 Fabrication of SERS substrates 55 3.2.5 SERS measurements 56 3.2.6 Data analysis 56 3.3 Results and Discussions 58 CHAPTER 70 Use of Aminothiophenol as an Indicator for the Analysis of Silver Nanoparticles in Consumer Products by Surface-Enhanced Raman Spectroscopy (SERS) 70 4.1 Introduction 72 4.2 Materials and Methods 75 4.2.1 Materials 75 4.2.2 Characterization of Ag NPs in the products 76 4.2.3 Determine Ag NPs in tested products 76 4.2.4 Conjugation of PATP onto Ag NPs 77 4.2.5 Detection of Ag NPs Using SERS and PATP-Ag NPs conjugation 77 4.2.6 Data Analysis 78 4.3 Results and Discussion 78 CHAPTER 88 Toxicity of Graphene Oxide on Intestinal Bacteria, and Caco-2 Cells 88 5.1 Introduction 89 5.2 Materials and Methods 91 5.2.1 Characterization of GO 91 5.2.2 Preparation of Bacterial strains 92 5.2.3 Effect of GO on the growth of E coli, L acidophilus, and B animalis 92 5.2.4 Mammalian cell study 93 5.2.5 MTT reduction assay 94 5.2.6 WST-8 proliferation assay 95 5.2.7 Scanning electron microscopy (SEM) analysis 95 v 5.2.8 Transmission electron microscopy (TEM) analysis 96 5.2.9 Statistical analysis 96 5.3 Results and Discussion 97 CHAPTER 110 Antibacterial Properties of Selenium Nanoparticles and Their Toxicity on Caco-2 Cells 110 6.1 Introduction 111 6.2 Materials and Methods 112 6.2.1 Chemicals, bacterial strains mammalian cells 112 6.2.2 Characterization of Se NPs 113 6.2.3 Preparation of bacterial strains 114 6.2.4 Synthesis of Se NPs 114 6.2.5 Effect of Se NPs on the growth of bacterial strains 115 6.2.6 Mammalian cell study 115 6.2.7 MTT reduction assay 116 6.2.8 WST-8 proliferation assay 116 6.2.9 Scanning electron microscopy (SEM) 117 6.2.10 Transmission electron microscopy (TEM) 117 6.2.11 Statistical analysis 118 6.3 Results and Discussion 118 6.3.1 Characterization of Se NPs 118 6.3.2 Antibacterial effects of Se NPs on pathogenic bacteria 121 6.3.3 Cytotoxic effect of Se NPs on Caco-2 cells 126 CHAPTER 130 Conclusions and Future Plans 130 Appendix 133 References 134 VITA 172 vi LIST OF TABLES Table Page Table Food and Food-related products that claim to contain nanoparticles Table Antibacterial effect of inorganic ENPs against different microorganisms 21 Table In vitro cytotoxicity effects of graphene materials and Se NPs 33 Table Band assignment of major peak in SER spectra form three pesticides* 64 Table Limit of detection of using G-Au-AuNRS substrate for detection of azinphosmethyl, carbaryl, and phosmet 69 Table Total concentration of silver and Ag NPs, average size, and the intensity of SERS spectra acquired from five commercial products 86 Table GO characteristics, values presented means ± SD from triplicate measurements 99 Table The zeta-potential values of selenium nanoparticles 120 Table Zeta potential values of bacteria strains 126 vii LIST OF FIGURES Figure Page Figure 2-1 Schematic of a surface-enhanced light scattering process (Schatz and others 2006; Alonso-González and others 2012) 14 Figure 3-1 Structure of substrates: (a) graphene-Au-AuNR (G-Au-AuNR); (b) grapheneAuNR (G-AuN); (c) Au-AuNR 55 Figure 4-1 SERS spectra of PATP, PATP mixed with AgNO3, PATP with 30 nm Ag NP3 79 Figure 4-2 Comparisons of enhancement effects from Ag NPs 81 Figure 4-3 Concentration-dependent SERS spectra (part of full scale) of Ag NPs with PATP (10 mg/mL) as an indicator (A); the linear relation between Raman intensity and Ag NPs concentration (B) 82 Figure 4-4 SERS spectra four five Ag NPs-containing dietary and antimicrobial products Negative controls were prepared using the solvent of PATP (methanol) 84 Figure 4-5 Characterization of Ag NPs in the dietary supplements and antimicrobial products (A) Dietary supplement; (B) nasal spray; (C) dietary supplement; (D) dietary supplement; (E) dietary supplement 85 Figure 5-1 UV–vis absorption spectrum of GO aqueous dispersion (A) FTIR spectrum of dried graphite oxide sample (B) 98 Figure 5-2 TEM images of GO aqueous dispersion 98 viii Peng D, Zhang J, Liu Q, Taylor EW 2007 Size effect of elemental selenium nanoparticles (Nano-Se) at supranutritional levels on selenium accumulation and glutathione S-transferase activity J Inorg Biochem 101(10):1457-63 Persson H, Købler C, Mølhave K, Samuelson L, Tegenfeldt JO, Oredsson S, Prinz CN 2013 Fibroblasts cultured on nanowires exhibit low motility, impaired cell division, and DNA damage Small 9(23):4006-16 Pham VT, Truong VK, Quinn MD, Notley SM, Guo Y, Baulin VA, Al Kobaisi M, Crawford RJ, Ivanova EP 2015 Graphene induces formation of pores that kill spherical and rod-shaped bacteria ACS nano 9(8):8458-67 Pi J, 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Science and Technology at the Shanghai Ocean University in 2010 She worked as a faculty in the Department of Food Analysis, Institute of Biotechnology and Food technology, Hochiminh University of Industry, Vietnam for two years Trang joined the Food Science Department of University of Missouri to pursue her Ph.D degree from fall 2012 to 2016 172 ... by the dean of the Graduate School, have examined the thesis entitled ANTIBACTERIAL, PLASMONIC AND TOXIC PROPERTIES OF ENGINEERED NANOPARTICLES Presented by Trang Ha Dieu Nguyen, a candidate for... Cytotoxicity of inorganic NPs to human cells 32 2.9 Mechanisms of cytoxocity of ENPS 35 2.10 Physical and chemical properties of NMs associated with nanotoxicity 37 2.11 Toxicity... DNA, and lipids can be damaged (Sanchez and others 2011) Release of metal ions and toxicity The hypothesis of releasing of metal ions by NPs is usually tested by comparing the toxicity of metal