Clemson University TigerPrints All Dissertations Dissertations 8-2013 Functionalized nanomaterials and their biological applications Pallavi Vedantam Clemson University, [email protected] Follow this and additional works at: https://tigerprints.clemson.edu/all_dissertations Part of the Microbiology Commons Recommended Citation Vedantam, Pallavi, "Functionalized nanomaterials and their biological applications" (2013). All Dissertations. 1194. https://tigerprints.clemson.edu/all_dissertations/1194 This Dissertation is brought to you for free and open access by the Dissertations at TigerPrints. It has been accepted for inclusion in All Dissertations by an authorized administrator of TigerPrints. For more information, please contact [email protected]. FUNCATIONALIZED NANOMATERIALS AND THEIR BIOLOGICAL APPLICATIONS A Dissertation Presented to the Graduate School of Clemson University In Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy Microbiology by Pallavi Vedantam August 2013 Accepted by: Dr Tzuen-Rong J. Tzeng, Committee Chair Dr Pu-Chun Ke Dr Lesly Temesvari Dr Ramakrishna Podila ABSTRACT Bionanomaterials have been used in drug delivery, cancer therapy and biodiagnosis of pathogens based on their size and surface functionalization. In this present work, different kinds of nanoparticles (NPs), their cellular interactions, cytotoxicity profiles, and finally role of gold nanoparticles (GNPs) in biodetection of E. coli was investigated. Firstly, cytotoxicity profiles of commercial, laser ablation, and green synthetic NPs were studied. Induction of apoptosis in cancer cells was found to be size dependent. The plain 80 nm GNPs and AgNPs enhanced toxic effects in cancer cells when compared to 20 nm ones. Apoptotic profiles of ALB- or FBS-coated NPs were significantly low in cancer cells when compared to plain NPs. The FBS-coated NPs were relatively bigger. Whereas, green synthetic NPs prepared from floral extracts of Tacoma stans and Tagetus erecta caused significant increase in cell membrane damage in cancer cells when compared to commercial GNPs. The phytochemicals in the extract were found to have a synergistic effect on pathogens like E. coli, Staphylococcus aureus and Enterococcus faecalis when used with antibiotics like tetracycline, ampicillin and vancomycin. This effect can be capitalized in developing NPs as effective drug carriers. Next, use of commercial GNPs as diagnostic tools to detect competitive binding in DU-145 in the presence of UTI-causing E. coli ORN178 was studied. GNPs functionalized with D-mannose (Mn) showed competitive binding between Mn-GNPs and E. coli ORN178 when presented together with DU-145 cells. Cytotoxicity assays of plain and Mn-GNPs showed significant decrease in viability of DU-145 cells. The ii plain/Mn 20 nm GNPs were taken up more by the cell when compared to the 200 nm ones. The protein-coated GNPs were found to be stable in culture medium. This competitive binding can be further developed to prevent/detect recurrent UTI in DU-45 cells. Lastly, primary and fine sugar specificity of fimbrial lectins of E.coli ORN178 and E. coli 13762 to D-mannose and Neuαc(α2-3)-Gal-(β1-4)Glc–Paa functionalized GNPs showed that E. coli ORN178 binds specifically only to Mn-GNPs and E. coli 13762 to the latter. Hence, adhesin-specific adhesion shows great potential for designing NPs to specifically bind to microorganisms as biodiagnostic tools. iii DEDICATION It is true that not even a blade of grass moves without His Will. This task would not be possible without the Divine blessings, guidance and grace of my most beloved guru and sakha, Bhagavan Sri Sathya Sai Baba. His Divine love helped me overcome every obstacle in this endeavor. I humbly dedicate this dissertation to Him and thank Him for everything. iv ACKNOWLEDGMENTS First and foremost, I would like to immensely thank Dr. Jeremy Tzeng for giving me an opportunity in pursuing PhD. But for his constant guidance and support I could not have completed this task. His continuous encouragement and sense of humor has definitely been instrumental in completing this project. More than anything his patience and outlook towards research has taught me a lot and shaped me as a better person. I would like to thank my committee members, Dr. Robert Latour for constructive feedback, Dr. Pu-Chun Ke for his invaluable suggestions, Dr. Lesly Temesvari for her inputs and Dr. Ramakrishna Podila for readily agreeing to be on my committee and for his stimulating scientific ideas. I would like to thank Dr. Wheeler for all the help he has extended during my education in the past six years. I am also thankful to Dr. Charlie Rice who had given me an opportunity to work in his lab. I am thankful to the staff of the Department of Biological Sciences for all their services. I am grateful to Dr. Thompson Mefford, Department of Material Sciences, Clemson University for help with instrumentation. I would like to extend my gratitude to Dr. R Sai Sathish, Sri Sathya Sai University for providing me the samples for research. I would like to express my gratitude to Dr. Guohui Huang who not only trained me in cell culture but also helped me design experiments and prepare the cells from time to time. v I would like to most importantly thank John Abercrombie who has been very patient, cooperative and resourceful in helping me overcome unforeseen obstacles in my research and teaching. I am grateful to all the past and present lab mates and undergraduate students who have helped me immensely. I am extremely thankful to almighty for gifting me friends like Dr. Pooja Puneet and Dr. Sally Wahba for being a part of my life from the beginning to the end of this educational pursuit. Their friendship has kindled optimism and enthusiasm in me to reach the finish line! I would like to express my heartfelt gratitude to my beloved sister, Indira Vedantam, for all her support and endless love. Lastly, I would like to thank my mother who has been the backbone of my life and co-advisor in checking the progress of my research from time to time! I am forever indebted to her for all her prayers, patience, compassion, blessings, sacrifices and love. vi LIST OF PUBLICATIONS 1. Vedantam, P., George Huang, and T. R. Jeremy Tzeng. “Size-Dependent Cellular Toxicity and Uptake of Commercial Colloidal Gold Nanoparticles in Du-145 Cells”. Cancer Nanotechnology, (2013): 4(1-3): 13-20. 2. Vedantam P., T. R. J. Tzeng, A. K. Brown, R. Podila, A. Rao, and K. Staley. “Binding of Escherichia coli to Functionalized Gold Nanoparticles”. Plasmonics 7, no. 2 (2012): 301-308. 3. J. Zeng, Y. Denga, P Vedantam, T. Tzeng, X. Xuan. “Magnetic separation of particles and cells in ferrofluid flow through a straight microchannel using two offset magnets”. Journal of Magnetism and Magnetic materials. 2013. dx.doi.org/10.1016/j.jmmm.2013.07.021 4. Podila, R., P. Vedantam, P. C. Ke, J. M. Brown, and A. M. Rao. "Evidence for Charge-Transfer-Induced Conformational Changes in Carbon Nanostructure–Protein Corona." The Journal of Physical Chemistry C 116, no. 41 (2012): 22098-22103. 5. Zeng, J., Chen, C., P. Vedantam., Tzeng, Tzuen-Rong, Xuan, Xiangchun. “Magnetic concentration of particles and cells in ferrofluid flow through a straight microchannel using attracting magnets”. Microfluidics and Nanofluidics, 2012: p. 1-7. 6. Patel, S., Daniel S., P. Vedantam, Tzuen-Rong T., Shizhi Q., and Xiangchun X. “Microfluidic Separation of Live and Dead Yeast Cells Using Reservoir-Based Dielectrophoresis.” Biomicrofluidics 6, no. 3 (2012): 034102-12. 7. Zeng, Jian, Chen Chen, P. Vedantam, Vincent Brown, Tzuen-Rong J Tzeng, and Xiangchun Xuan. “Three-Dimensional Magnetic Focusing of Particles and Cells in Ferrofluid Flow through a Straight Microchannel.” Journal of Micromechanics and Microengineering 22, no. 10 (2012): 105018. 8. Zhu, J., Robert C., Gyunay K., P. Vedantam, Tzuen-Rong J., and Xiangchun X. “Continuous-Flow Particle and Cell Separations in a Serpentine Microchannel Via Curvature-Induced Dielectrophoresis.” Microfluidics and Nanofluidics 11, no. 6 (2011): 743-752. 9. Badal, S., S. A. Williams, G. Huang, S. Francis, P. Vedantam, O. Dunbar, H. Jacobs, T. J. Tzeng, J. Gangemi, and R. Delgoda. “Cytochrome P450 1 Enzyme Inhibition and Anticancer Potential of Chromene Amides from Amyris Plumieri.” Fitoterapia 82, no. 2 (2011): 230-236. vii TABLE OF CONTENTS Page TITLE PAGE .................................................................................................................... i ABSTRACT ..................................................................................................................... ii DEDICATION ................................................................................................................ iv ACKNOWLEDGMENTS ............................................................................................... v LIST OF PUBLICATIONS ........................................................................................... vii LIST OF TABLES .......................................................................................................... xi LIST OF FIGURES ....................................................................................................... xii CHAPTER - I. REVIEW OF LITERATURE ........................................................................ 1 Synthesis .................................................................................................. 1 Ligands ..............................................................................................