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

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.

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

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.

Hypothesis The two delivery techniques will be

comparable, through both qualitative and quantitative means

Variables• Method of drug deliveryIndependen

t

• 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

Equipment Electrospinning apparatus Scanning electron microscope (SEM) Homogenizer Incubator Environmental chamber Spectrophotometer

Materials• Bovine Serum Albumin• Alcohol• A. tumefacians• Potato strips• Diffusion assays• tetracycline and ampicillin

Preparation of albumin nanodropletsEmulsification• 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

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

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

Antibiotic loading - nanofibresAntibiotics mixed in albumin

solution Homogenous solution Hypothesis that spinning solution will

result in the non polymer antibiotics also being spun

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

Timeline (HCI)Form droplets

w/ specific concentration

and temp.

Load droplets with antibiotics

Test droplets Send for characterization

Examine results and

modify original solution

Timeline (AOS)Form solution with specific

concentration

Spin solution

Test fibersSend for characterization

Examine results and

modify original solution

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

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.

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

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.