Progress 10/01/11 to 09/30/16
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Several undergraduate and graduate students have participated in this project and received training. How have the results been disseminated to communities of interest?10 peer-reviewed journal articlesand several meeting abstracts have been published. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
We successfully synthesized heterobifunctional polymer coated magnetic nanoparticles that have nitroDOPA as a stable anchoring agent and were bio-functionalized with sialic-acid glycoconjugate (Neu5Ac(α2-3)Gal(β1-4)Glcβ-sp) (GM3-MNPs) using click chemistry. Our GM3-MNPs specifically interacted only with ETEC strain EC K99 as confirmed through fluorescence microscopy and transmission electron microscopy. When such EC K99 and GM3-MNPs nanoparticle mixtures were exposed to alternative magnetic fields, a 3-log reduction in CFU of EC K99 was achieved due to GM3-MNPs mediated energy transfer. Moreover, we have demonstrated that when mixtures of EC K99 (target bacteria) and EC O157:H7 (non-target bacteria) were subjected to such GM3-MNPs mediated energy transfer, a 2-log reduction in CFU of EC K99 was observed but only and a limited 1-log reduction in CFU of EC O157 (K88) was observed in a heterogeneous environment. Further, the preliminary cytotoxicity assay results on human cell-line proved the highly biocompatible nature of our nanoparticles system. Thus, our proof-of-concept nanoparticle system can effectively serve as novel non-antibiotic multivalent carriers, which could find applications in detection and capturing of pathogenic multi-drug resistant bacterial strains from active physiological body fluids. Our systems can especially reduce/treat gastro-intestinal tract infections caused by ETEC pathogens in farm animals and humans since specific bacterial-nanoparticle aggregates can be effectively flushed out from the body system because of high peristaltic flows without disturbing the normal gut microflora that is usually destroyed when antibiotics are used. This system can also be employed as potent anti-adhesion agents that can block/inhibit specific cellular responses by competitively preventing the attachment of bacterial pathogens onto specific eukaryotic cell-surface receptors and thereby reducing the infection load.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
1. Time domain detection and differentiation of single particles and cells with a radio frequency interferometer
Z Wang, Y Raval, TR Tzeng, B Booth, B Flaherty, D Peterson, J Moore, D Rosenmann, R Divan, G Yu, Pingshan Wang
2016 IEEE Topical Conference on Biomedical Wireless Technologies, Networks, and Sensing Systems (BioWireleSS), Jan. 24; 77-80
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
2. Highly stable multi-anchored magnetic nanoparticles for optical imaging within biofilms
R.C. Stone, B.D. Fellows, B.Qi, D. Trebatowski, B. Jenkins, Y. Raval, T.R. Tzeng, T.F. Bruce, T. McNealy, M.J. Aust6in, T.C. Monson, D.L. Huber, O.T. Mefford
Journal of Colloid and Interface Science, DOI: 10.1016/j.jcis2015.08.12. Dec. 1; 459: 175-182
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Progress 10/01/14 to 09/30/15
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?Journal article and conference posters. What do you plan to do during the next reporting period to accomplish the goals?The mechanisms of killing using bacterial adhesin-specific superparamagnetic iron oxide nanoparticles and potential toxicities associated with the use of these nanoparticles will be characterized using various biological assays.
Impacts
What was accomplished under these goals?
We successfully synthesized heterobifunctional polymer coated magnetic nanoparticles that have nitroDOPA as a stable anchoring agent and were bio-functionalized with sialic-acid glycoconjugate (Neu5Ac(α2-3)Gal(β1-4)Glcβ-sp) (GM3-MNPs) using click chemistry. Our GM3-MNPs specifically interacted only with ETEC strain EC K99 as confirmed through fluorescence microscopy and transmission electron microscopy. Also, a 2-log reduction in CFU of EC K99 was achieved due to GM3-MNPs induced bacterial aggregation. Moreover, intracellular ATP assays demonstrated that the 2-log reduction in CFU of EC K99 was not due to inherent toxicity of the nanoparticles. Further, the preliminary cytotoxicity assay results on human cell-line proved the highly biocompatible nature of our nanoparticles system. Thus, our proof-of-concept nanoparticle system can effectively serve as novel non-antibiotic multivalent carriers, which could find applications in detection and capturing of pathogenic multi-drug resistant bacterial strains from active physiological body fluids. Our systems can especially reduce/treat gastro-intestinal tract infections caused by ETEC pathogens in farm animals and humans since specific bacterial-nanoparticle aggregates can be effectively flushed out from the body system because of high peristaltic flows without disturbing the normal gut microflora that is usually destroyed when antibiotics are used. This system can also be employed as potent anti-adhesion agents that can block/inhibit specific cellular responses by competitively preventing the attachment of bacterial pathogens onto specific eukaryotic cell-surface receptors and thereby reducing the infection load.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Synthesis and application of glycoconjugatefunctionalized magnetic nanoparticles as potent anti-adhesion agents for reducing enterotoxigenic Escherichia coli infections
Nanoscale, 2015, 7, 8326-8331
Yash S. Raval, Roland Stone, Benjamin Fellows, Bin Qi, Guohui Huang, O. Thompson Mefford, and Tzuen-Rong J. Tzeng
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Progress 10/01/13 to 09/30/14
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest? Journal article and conference posters. What do you plan to do during the next reporting period to accomplish the goals? Continue to optimize adhesin-specific nanoparticles/devices for inactivation/detection of drug-resistant pathogens.
Impacts
What was accomplished under these goals?
We successfully synthesized heterobifunctional polymer coated magnetic nanoparticles that have nitroDOPA as a stable anchoring agent and were bio-functionalized with sialic-acid glycoconjugate (Neu5Ac(α2-3)Gal(β1-4)Glcβ-sp) (GM3-MNPs) using click chemistry. The GM3-MNPs were characterized by employing different techniques and their adhesin specificity was determined using aggregation assays. Our GM3-MNPs specifically interacted only with ETEC strain EC K99 as confirmed through fluorescence microscopy and transmission electron microscopy. Also, a 2-log reduction in CFU of EC K99 was achieved due to GM3-MNPs induced bacterial aggregation. Moreover, intracellular ATP assays demonstrated that the 2-log reduction in CFU of EC K99 was not due to inherent toxicity of the nanoparticles. Thus, our proof-of-concept nanoparticle system can effectively serve as novel non-antibiotic multivalent carriers, which could find applications in detection and capturing of pathogenic multi-drug resistant bacterial strains from active physiological body fluids. Our systems can especially reduce/treat gastro-intestinal tract infections caused by ETEC pathogens in farm animals and humans since specific bacterial-nanoparticle aggregates can be effectively flushed out from the body system because of high peristaltic flows without disturbing the normal gut microflora that is usually destroyed when antibiotics are used. This system can also be employed as potent anti-adhesion agents that can block/inhibit specific cellular responses by competitively preventing the attachment of bacterial pathogens onto specific eukaryotic cell-surface receptors and thereby reducing the infection load. Furthermore, this nanoparticle system can also be utilized for targeted magnetic hyperthermia treatment of bacterial infections, especially those that are resistant to multiple antibiotics. Future work involves utilizing multi-anchored functional groups for improving stability of nanoparticles in biological fluids and performing anti-adhesion assays in cell-lines and on small animals.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Exploiting Magnetic Asymmetry to Concentrate Diamagnetic Particles in Ferrofluid Microflows
JJ Wilbanks, G Kiessling, J Zeng, C Zhang, TR Tzeng, X Xuan
Journal of Applied Physics 115 (4), 044907-044907-7
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Progress 01/01/13 to 09/30/13
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest? Journal articles and conference posters and presentations. What do you plan to do during the next reporting period to accomplish the goals? We would like to fabricate and characterize additional nanoparticles for inactivation of other food borne pathogens, e.g.,Campylobacter jejuni. We would also like to explore the use of carbohydrate-coated magnetic nanoparticles for microfluidic bacterial separation from blood.
Impacts
What was accomplished under these goals?
Adherence is among the earliest event in many bacterial infections. These interactions are required for extracellular colonization and/or internalization. This colonization is usually mediated by bacterial adhesins on the surface of bacteria that recognize and bind to specific receptor moieties of host cells. This adhesin-receptor binding event could activate cascades of signal transduction important in the pathogenic process and host-defense. Understanding of this adhesin-receptor interaction is of great value in order to develop effective prevention, detection, diagnostic, and treatment methods. The use of carbohydrates as capturing agents in biosensor development represents a more specific, stable, and economical approach than the conventional immunoassays or PCR detection methods that are more costly and incapable of differentiating non-viable from viable targets. By the same token, carbohydrates functionalized nanoparticles could mimic host cell surface and mediate bindings of pathogens to such nanoparticles. Such interactions could be utilized to physically remove pathogens from hosts without the use of antibiotics. In this present work, the PI and his team have functionalized nanoparticles with receptor molecules recognized by specific bacterial adhesins and demonstrated that iron oxide nanoparticles functionalized with Neu5Ac(α2-3)-Gal-(β1-4)Glc- could bind specifically to E. coli K99, the main causative agent of bloody diarrhea in young calves, lambs, and pigs, to form nanoparticle-bacterium aggregates. When subjected to magnetic fields, such aggregated bacteria could be killed via localized hyperthermia without compromising the viability of non-targets. The cytotoxicity of such functionalized nanoparticles has been demonstrated to be minimal. In addition, the cytotoxicity profiles of commercial, laser ablation, and green synthetic nanoparticles were studied. The PI and his team have also demonstrated the manipulation of particles in microfluidic devices. Such devices will lay the foundation for development of biosensors.
Publications
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2013
Citation:
Functionalized Nanomaterials and Their Biological Applications, Pallavi Vedantam, August, 2013
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Magnetic concentration of particles and cells in ferrofluid flow through a straight microchannel using attracting magnets
J Zeng, C Chen, P Vedantam, TR Tzeng, X Xuan
Microfluidics and nanofluidics 15 (1), 49-55
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Development of luminescent pH sensor films for monitoring bacterial growth through tissue
F Wang, Y Raval, H Chen, TRJ Tzeng, JD DesJardins, JN Anker
Advanced healthcare materials
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Magnetic separation of particles and cells in ferrofluid flow through a straight microchannel using two offset magnets
J Zeng, Y Deng, P Vedantam, TR Tzeng, X Xuan
Journal of Magnetism and Magnetic Materials 346, 118-123
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Size-dependent cellular toxicity and uptake of commercial colloidal gold nanoparticles in DU-145 cells
P Vedantam, G Huang, TRJ Tzeng
Cancer Nanotechnology 4 (1-3), 13-20
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: The objectives of this project are to 1. Develop new technologies for characterizing fundamental nanoscale processes 2. Construct and characterize self-assembled nanostructures 3. Develop devices and systems incorporating microfabrication and nanotechnology 4. Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems 5. Produce education and outreach materials on nanofabrication, sensing, systems integration and application risk assessment. The PI and his team have fabricated several nanomaterials/devices to study nanoscale phenomena for applications in pathogen detection/inactivation and risk assessment. The findings have been disseminated to the scientific community via seminars, international conference, and manuscripts as well as a developed web site. PARTICIPANTS: Tzuen-Rong Tzeng, associate professor serving as the PI on this project, is responsible for overseeing the research activities, research designs, data analysis and communication. Pallavi Vedantam, PhD candidate, who serves as an investigator is responsible for designing and carrying out the designed experiments for characterizing adhesin-specific gold nanoparticles for binding to foodborne pathogens and the isolation and detection of pathogens in microfluidic channels. Mr. Yash Raval, PhD graduate student, who serves as an investigator is responsible for designing and carrying out the designed experiments for adhesin-specific iron-oxide nanoparticle mediated hyperthermia inactivation of foodborne pathogens. List of Collaborators: Dr. Xiangchun Xuan, Assistant Professor, Mechanical Engineering, Clemson University, collaborated on projects related to microfluidic manipulation of particles. xcxuan@clemson.edu Dr. Thompson Mefford, Assistant Professor, Materials Science & ENGRG, Clemson University, collaborated on projects related to adhesin-specific iron-oxide nanoparticles. mefford@clemson.edu Dr. Elizabeth Carraway, Professor, Environmental ENGR & Earth Sci, Clemson University. ecarraw@clemson.edu Dr. Mark Schlautman, Professor, Environmental ENGR & Earth Sci, Clemson University, mschlau@clemson.edu TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Adherence is among the earliest events in many bacterial infections. These interactions are required for extracellular colonization and/or internalization. This colonization is usually mediated by bacterial adhesins on the surface of bacteria that recognize and bind to specific receptor moieties of host cells. This adhesin-receptor binding event could activate cascades of signal transduction important in the pathogenic process and host-defense. Understanding of this adhesin-receptor interaction is of great value in order to develop effective prevention, detection, diagnostic, and treatment methods. Nanotechnology offers new opportunities for the nanoscale investigation of adhesin-receptor interactions. We envision that adhesin specific nanoparticles and biosensors will break new ground in study of pathogen-host interactions, drug discovery, drug delivery, and biosensing. To achieve such goals, the fundamentals of adhesin-receptor interactions must be examined. In this present work, the PI and his team have functionalized nanoparticles with receptor molecules recognized by specific bacterial adhesins to demonstrate that such nanoparticles could be utilized to eliminate pathogens from host without the need for antibiotics that many pathogens are now resistant to. The PI and his team have also utilized the same principle to develop highly sensitive biosensors capable of detecting the targeted pathogens in low number. The PI has received a resource grant from the Consortium for Functional Glycomics allowing the PI to tap into their carbohydrate resources enabling the PI to functionalize various nanomaterials for the proposed studies. In this report period, the PI has requested 3 carbohydrates, i.e., Tr117, Tr32, and Tr33, for functionalization of iron-oxide nanoparticles and is currently evaluating their antimicrobial property against specific foodborne pathogen via localized hyperthermia. The use of carbohydrates as capturing agents in biosensor development represents a more specific, stable, and economical approach than the conventional immunoassays or PCR detection methods that are more costly and incapable of differentiating non-viable from viable targets.
Publications
- 1. Zeng J., C. Chen, P. Vedantam, T.-R. Tzeng, X. Xuan (2012) Magnetic concentration of particles and cells in ferrofluid flow through a straight microchannel using attracting magnets. Microfluidics and Nanofluidics, Epub ahead of print, Dec. 2012
- 2. Zeng, J., C. Chen, P. Vedantam, V. Brown, T.-R. Tzeng, X. Xuan (2012). Three-dimensional magnetic focusing of particles and cells in ferrofluid flow through a straight microchannel. Journal of Micromechanics and Microengineering, Epub ahead of print: 22, 105018
- 3. Patel, S., D. Showers, P. Vedantam, V. Brown, T.-R. Tzeng, Q. Shizhi, X. Xuan (2012). Microfluidic separation of live and dead yeast cells using reservoir-based dielectrophoresis (rDEP). Biomicrofluidics, Epub ahead of print, Vol. 6, Issue 3, Sept. 2012
- 4. Vedantam, P., T.-R. Tzeng (2012). Binding of Escherichia coli to functionalized gold nanoparticles. Plasmonics, Epub ahead of print, Vol. 7, Number 2, 301-308
- 5. Xuan, P., Y. Zhang, T.-R. Tzeng, X.F. Wan, F. Luo (2012). A quantitative structure-activity relationship (QSAR) study on glycan array data to determine the specificities of glycan-binding proteins, Glycobiology, Vol. 2, Issue 4, 552-560
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: The objectives of this project are to 1. Develop new technologies for characterizing fundamental nanoscale processes 2. Construct and characterize self-assembled nanostructures 3. Develop devices and systems incorporating microfabrication and nanotechnology 4. Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems 5. Produce education and outreach materials on nanofabrication, sensing, systems integration and application risk assessment. The PI and his team have fabricated several nanomaterials/devices to study nanoscale phenomena for applications in pathogen detection/inactivation and risk assessment. The findings have been disseminated to the scientific community via seminars, international conference, and manuscripts as well as a developed web site. PARTICIPANTS: Dr. Tzuen-Rong Jeremy Tzeng, PI, Assistant Professor, Biological Sciences, Clemson University; Dr. Xiangchun Xuan, Co-investigator, Assistant Professor, Mechanical Engineering, Clemson University; Dr. Feng Luo, Co-investigator, Assistant Professor, School of Computing, Clemson University; Dr. Elizabeth Carraway, Co-investigator, Associate Professor, environmental Engineering & Earth Science, Clemson University; Dr. Mark Schlautman, Co-investigator, Associate Professor, environmental Engineering & Earth Science, Clemson University; Dr. Nader Jalili, Co-investigator, Associate Professor, Mechancial & Industrial Engineering, Northeastern University; Ms. Pallavi Vedantam, PhD candidate, Biological Sciences, Clemson University. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Adherence is among the earliest event in many bacterial infections. These interactions are required for extracellular colonization and/or internalization. This colonization is usually mediated by bacterial adhesins on the surface of bacteria that recognize and bind to specific receptor moieties of host cells. This adhesin-receptor binding event could activate cascades of signal transduction important in the pathogenic process and host-defense. Understanding of this adhesin-receptor interaction is of great value in order to develop effective prevention, detection, diagnostic, and treatment methods. Nanotechnology offers new opportunities for the nanoscale investigation of adhesin-receptor interactions. We envision that adhesin specific nanoparticles and biosensors will break new ground in study of pathogen-host interactions, drug discovery, drug delivery, and biosensing. To achieve such goals, the fundamentals of adhesin-receptor interactions must be examined. In this present work, the PI and his team have functionalized nanoparticles with receptor molecules recognized by specific bacterial adhesins to demonstrate that such nanoparticles could be utilized to eliminate pathogens from host without the need for antibiotics that many pathogens are now resistant to. The PI and his team have also utilized the same principle to develop highly sensitive biosensors capable of detecting the targeted pathogens in low number. The PI has received a resource grant from the Consortium for Functional Glycomics allowing the PI to tap into their carbohydrate resources enabling the PI to functionalize various nanomaterials for the proposed studies. The use of carbohydrates as capturing agents in biosensor development represents a more specific, stable, and economical approach than the conventional immunoassays or PCR detection methods that are more costly and incapable of differentiating non-viable from viable targets.
Publications
- Vedantam, P., K. Staley, T.-R. Tzeng (2011). Binding of functionalized gold nanoparticles to Escherichia coli. Plasmonics, accepted
- Xuan, P., Y. Zhang, T.-R. Tzeng, X.F. Wan, F. Luo (2011). A quantitative structure-activity relationship (QSAR) study on glycan array data to determine the specificities of glycan-binding proteins, Glycobiology, accepted
- Zhu, J., R.C. Canter, G. Keten, P. Vedantam, T.J. Tzeng, and X. Xuan, (2011). Continuous-flow particle and cell separations in a serpentine microchannel via curvature-induced dielectrophoresis, Microfluidics and Nanofluidics, Epub ahead of print, July 07, 2011
- Tzeng, T.-R., Y.R. Cheng, R. Saeidpourazar, S.S. Aphale, N. Jalili, (2011). Adhesin-Specific Nanomechnical Cantilever Biosensors for Detection of Microorganisms, Journal of Heat Transfer, 133, Issue 1 Adhesin-Specific Nanoparticles for Removal of Enteropathogens, International Symposium on Applications of Nanotechnology and Biosensors in Agriculture and Food, Zhejiang University, Hangzhou, China, April 14-16, 2011
- SHOWERS, D., BROWN, V., LIANG, L., TZENG, T. J., XUAN, X. Dielectrophoretic Separation of Live and Dead Yeast Cells in Microfluidic Reservoirs, 64th Annual Meeting of the APS Division of Fluid Dynamics, Baltimore, MD, Log number DFD11-2011-000123 (November 2011)
- ZENG, J., TZENG, T. J., XUAN, X. Diamagnetic Cell Focusing in Ferrofluid Microchannel Flows, 64th Annual Meeting of the APS Division of Fluid Dynamics, Baltimore, MD, Log number DFD11-2011-000119 (November 2011)
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