Enhancing Antibacterial Efficacy using Protein Nanoparticles Leslie Tan Zheng Yu Tan Jing Chong Erik...
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Enhancing Antibacterial Efficacy using Protein Nanoparticles
Leslie Tan Zheng Yu Tan Jing Chong Erik WarnquistVarun Kulkarni
Retrieved from: http://www.eng.uci.edu/files/images/gallery/Protein_Nanoparticle_Structure.jpg
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Introduction Pesticides are used to eradicate
Agrobacterium tumefaciens High percentage of pesticide does not
reach the target species. Result in water and soil pollution. Threatens biodiversity.
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Introduction Usage of nanoparticles as drug carrier
for pesticides Increase in therapeutic efficacy Increasing localisation to diseased sites Decrease in side effect Protein Nanoparticle are biodegradable,
metabolisable and non-antigenic Does not accumulate in tissue
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Objective To compare the effectiveness of
antibiotic loaded albumin nanodroplets against antibiotic loaded albumin nanofibre on A. tumefaciens, grown both in vitro and in vivo.
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Hypothesis The two delivery techniques will be
comparable, through both qualitative and quantitative means
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Variables
• Method of drug deliveryIndependent
• Efficacy of drug delivery systemDependent
• Type of bacteria (A.tumefaciens)
• Volume and types of antibiotic - tetracycline and ampicillin
• Agrobacterium volume• Sizes of potato strips• Temperature and humidity
Controlled / constant
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Equipment Electrospinning apparatus Scanning electron microscope (SEM) Homogenizer Incubator Environmental chamber Spectrophotometer
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Materials• Bovine Serum Albumin• Alcohol• A. tumefacians• Potato strips• Diffusion assays• tetracycline and ampicillin
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Preparation of albumin nanodroplets
Emulsification• Aqueous Bovine Serum Albumin is turned
into an emulsion at room temperature and in oil
• A homogenizer is used to make the emulsion homogeneous. There is a high dispersion of particles
• Emulsion is added to pre-heated oil• Albumin nanoparticles are separated by
desolvating agent eg. Alcohol
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Preparation of albumin nanofibers
Electrospinning• Solution inside a syringe exposed to
initial electric field• Electric field increases in charge• Point is reached where attractive forces
of charges exceeds surface tension• The fibers are projected onto a
grounded collector
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Antibiotic loading - nanodroplets• Incubating nanoparticles in antibiotic
solution• Antibiotic contained in nanoparticles• Done at protein's isoelectric point
Minimum solubility and maximum absorption BSA: pH of 4.4• Larger amount of antibiotic loaded• Antibiotic entrapment efficacy measured
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Antibiotic loading - nanofibresAntibiotics mixed in albumin
solution Homogenous solution Hypothesis that spinning solution will
result in the non polymer antibiotics also being spun
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Effectiveness of antibiotic-loaded nanoparticles• Protein nanoparticles digested by
proteases to release antibiotics• Antibiotic-loaded nanoparticles are
subjected to:• A.tumefacians agar plates discs• A.tumefacians-potato strips
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Timeline (HCI)Form droplets
w/ specific concentration
and temp.
Load droplets with antibiotics
Test droplets Send for characterization
Examine results and modify
original solution
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Timeline (AOS)Form solution with specific
concentration
Spin solution
Test fibersSend for characterization
Examine results and modify
original solution
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References Buschle-Diller, G., Cooper, J., Xie, Z., Wu, Y.,
Waldrup, J., & Ren, X. (2007). Release of antibiotics from electrospun bicomponent fibers. Cellulose, 14(6), 553- 562
Collins, A. (2001). Agrobacterium tumefaciens. Department of Plant Pathology, University of North Carolina State. Retrieved September 19, 2010 from: http:/www.cals.ncsu.edu/course/pp728/Agrobacterium/Alyssa_Collins_profile.htm
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Frenot, A., & Chronakis, I.S. (2003). Polymer nanofibers assembled by
electrospinning. Current Opinion in Colloid and Interface Science, 8(1), 64-75.
Hyuk, Y.S., Taek, G.K., & Park, T.G. (2009). Surface-functionalized electrospun
nanofibers for tissue engineering and drug delivery. Advanced Drug Delivery Reviews, 61(12), 1033-1042.
Jahanshahi, M. & Babaei, Z. (2008). Protein nanoparticle: A unique system as drug delivery vehicles. African
Journal of Biotechnology, 7(25), 4926-4934.
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Knee, M., & Nameth, S. (2007). Horticulture and Crop Science: Bacteria. The Ohio State University, Horticulture Department. Retrieved September 12, 2010 from : http://www.hcs.ohio-state.edu/hcs300/bact.htm
Kratz, F. (2008). Albumin as a drug carrier: Design of prodrugs, drug conjugates and nanoparticles. Journal of Controlled Release, 132(3), 171-183.
McManus, P. (2007). Antibiotic Use in Plant Disease Control. Fruit Pathology: University of Wisconsin-Madison. Retrieved September 13, 2010 from: http://www.plantpath.wisc.edu/fpath
/antibiotic-use.htm
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M.R., Jahanshahi, M., & Najafpour, G.D. (2006). Production of biological nanoparticles from bovine serum albumin for drug delivery. African Journal of Biotechnology, 5(20),
1918-1923.