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Copper/N-Acetylcysteine Coated Iron Oxide Nanoparticles Synthesis, Characterization, and Antimicrobial Activities

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Copper/N-Acetylcysteine Coated Iron Oxide Nanoparticles Synthesis, Characterization, and Antimicrobial Activities University of Central Florida STARS Electronic Theses and Dissertations, 2020- 2020 Copper/N-acetylcysteine Coated Iron Oxide Nanoparticles Synthesis, Characterization, and Antimicrobial Activities Danya Belnour University of Central Florida Part of the Medical Biotechnology Commons Find similar works at: https://stars.library.ucf.edu/etd2020 University of Central Florida Libraries http://library.ucf.edu This Masters Thesis (Open Access) is brought to you for free and open access by STARS. It has been accepted for inclusion in Electronic Theses and Dissertations, 2020- by an authorized administrator of STARS. For more information, please contact [email protected]. STARS Citation Belnour, Danya, "Copper/N-acetylcysteine Coated Iron Oxide Nanoparticles Synthesis, Characterization, and Antimicrobial Activities" (2020). Electronic Theses and Dissertations, 2020-. 174. https://stars.library.ucf.edu/etd2020/174 COPPER/ N-ACETYLCYSTEINE COATED IRON OXIDE NANOPARTICLES SYNTHESIS, CHARACTERIZATION, AND ANTIMICROBIAL ACTIVITY By DANYA BELNOUR B.S. University of Central Florida, 2018 A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in the Burnett School of Biomedical Sciences in the College of Medicine at the University of Central Florida Orlando, Florida Summer Term 2020 Major Professor: Swadeshmukul Santra © 2020 Danya Belnour ii ABSTRACT In recent years, there is a growing interest in developing metal based antimicrobial nanomaterials suitable for agricultural and biomedical applications. For centuries, Copper (Cu) biocide has been used for protecting a wide variety of crops from devastating bacterial and fungal diseases. However, prolonged and aggressive use of Cu led to the development of resistance and accumulation in soil. The latter has been linked to aquatic toxicity due to soil Cu leaching. Furthermore, copper build up in soil causes phytotoxicity and reduces uptake of micronutrient Zn through the root system. In biomedical field, Cu has been historically used as an antimicrobial agent in wound dressing. In healthcare facilities, Cu based touch surfaces are shown to significantly reduce antimicrobial infection rates. Emerging biomedical applications include wound healing, sensing and even potential use of nano-Cu as cancer therapeutic. However, Cu cytotoxicity is a serious concern. There is a strong need for developing advanced Cu based composite material that will retain antimicrobial properties as reduced Cu load until a suitable alternative becomes available. In this thesis, the objective is to develop a Cu coated Iron Oxide nanocomposite. The idea is to distribute Cu over the high surface area of biocompatible Iron Oxide nanoparticle (IONP) to improve Cu bioavailability. In the design of Cu-IONP nanocomposite, we have introduced N-Acetyl Cysteine (NAC, an antioxidant biomolecule) that anchors IONP to Cu. The composite was synthesized using a co-precipitation technique. Characterization of the Cu/NAC-IONP nanocomposite was done using Atomic Absorption Spectroscopy (AAS), Dynamic Light Scattering, Fourier Transform Infrared Spectroscopy (FTIR), Scanning electron microscopy and X-Ray Photoelectron Spectroscopy. Characterization iii results support the formation of composite and NAC as a bidentate ligand conjugating Cu to IONP. Antimicrobial activity of Cu/NAC-IONP nanocomposite was studied using ASTM published protocol. The Cu/NAC-IONP nanocomposite shows higher Colony Forming Unit Percent reduction when compared with NAC-Iron Oxide and no treatment. This suggests that antimicrobial activity of Cu is retained in the nanocomposite. Additionally, AAS study revealed an interesting property of the nanocomposite that the Cu release is strongly dependent on incubation temperature. Cu ion release is increased with the increase in incubation temperature. This new finding may lead to design of a delivery system where Cu release can be controlled by tuning temperature. iv This thesis is dedicated to my loving parents Iman Tunali and Khalid Banur, and my siblings Mohammed, Seraj and Shehab Belnour. Your infinite love and support are the drive the keeps me going. I am grateful for each one of you. v ACKNOWLEDGMENTS First and Foremost, my deepest gratitude goes to Dr. Maria Campos. Her sincere mentorship through explaining concepts in material chemistry, recommendation of experiment design, and data analysis training are the reason this project is possible. Thank you for continues support, guidance, and advice. I am honored and proud to have been one of your mentees. Also, my sincerest and deepest thanks to my thesis adviser Dr. Swadeshmukul Santra for this amazing opportunity and guidance. I am grateful to have had the opportunity to learn from you and be part of your group. Thank you to my thesis committee member Dr. Ellen Kang and Dr. Michal Masternak for their kindness, support and valuable advice throughout the projects. In addition, I would like to thank Dr. Hajeewaka Mendis for Microbiology training, and Dr. Matthew Rex for Atomic Absorption Spectroscopy and FTIR instrument training. I am thankful to all my lab mates, teachers, and friends for their support and encouragements through the years. Finally, thank you to Material characterization Facility (MCF) and NanoScience Technology Center (NSTC) at the University of Central Florida for characterization instruments access and instrument training. I would like to acknowledge the Citrus Research and Development Foundation (CRDF project # 18-020) for providing partial support and the UCF Materials Innovation for Sustainable Agriculture (MISA) center for the technical support. vi TABLE OF CONTENTS LIST OF FIGURES ....................................................................................................................... ix LIST OF TABLES ........................................................................................................................ xii LIST OF ABBREVIATIONS ...................................................................................................... xiii CHAPTER ONE: INTRODUCTION ............................................................................................. 1 1.1: Nanotechnology in Agriculture ........................................................................................... 1 1.2: Antimicrobial Nanomaterials for Agriculture Purposes ...................................................... 2 1.3: Nanomaterial Design ........................................................................................................... 4 1.3.1: Iron Oxide Nanomaterial .............................................................................................. 5 1.3.2: Copper and N-acetylcysteine Nanomaterial Coating .................................................... 7 CHAPTER TWO: NANOMATERIALS CHARACTERIZATION ............................................ 12 2.1: Characterizations Techniques ............................................................................................ 12 2.1.1: Atomic Absorption Spectroscopy (AAS) ................................................................... 13 2.1.2: Dynamic Light Scattering (DLS) ................................................................................ 14 2.1.3: Fourier Transform Infrared Spectroscopy (FTIR) ...................................................... 15 2.1.4: Scanning Electron Microscopy (SEM) ....................................................................... 15 2.1.5:X-ray Photoelectron Spectroscopy (XPS) ................................................................... 16 CHAPTER THREE: METHODOLOGY ..................................................................................... 17 3.1: Nanomaterial Synthesis Procedure .................................................................................... 17 3.1.1: Iron Oxide Nanoparticle (IONP) Preparation ............................................................. 17 3.1.2: N-acetylcysteine-Iron Oxide Nanoparticle (NAC-IONP) Preparation ....................... 18 vii 3.1.3: Copper/N-acetylcysteine- Iron oxide Nanoparticle (Cu/NAC-IONP) Preparation .... 19 3.2: Antimicrobial Studies ........................................................................................................ 23 3.2.1: Determination of Antimicrobial Activity of Nanomaterials ....................................... 23 3.2.1.A: Antimicrobial Activity of NAC-IONP and Cu/NAC-IONP on Escherichia coli 24 3.2.1.B: Antimicrobial Activity of NAC-IONP and Cu/NAC-IONP on Pseudomonas syringea ............................................................................................................................. 25 3.2.2: Quantification of Copper Release ................................................................................... 26 CHAPTER FOUR: RESULTS & DISCUSSION......................................................................... 28 CHAPTER FIVE: CONCLUSION............................................................................................... 55 APPENDIX: SUPPLEMENTERY INFORMATION .................................................................. 56 LIST OF REFERENCES .............................................................................................................. 61 viii LIST OF FIGURES Figure 1:Nanomaterial design and possible interaction of molecules. A: Iron oxide (IONP), B: N- Acetylcysteine-Iron Oxide (NAC-IONP) and Copper/N-acetylcysteine-Iron Oxide (Cu/NAC-IONP). “Created with BioRender.com.” .........................................................
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