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doctoral thesis

Preparation and Analysis of Bacteria Surface-Modified with Polymer Nanoparticles

Bader, Lisa Patricia Elisabeth  
2022

Cell-based drug carriers present an interesting approach to improve the targeted delivery of therapeutic drugs in vivo. Specifically, bacterial cells possess interesting properties such as motility and sensing that allow some strains to selectively target tumor areas. Immobilization of abiotic materials on the cell surface of such bacterial cells can therefore enable the delivery of therapeutic compounds at the desired location. The work presented in this thesis explores the fabrication and characterization of bacteria-nanoparticle biohybrids as potential carriers for drug delivery applications. Chapter 1 presents a review of the scientific literature that covers the different approaches to immobilize abiotic materials on the surface of bacteria. It highlights the advantages of using cells as carriers for the transport of therapeutics. An effective drug delivery agent should present great motility, viability, biodistribution and biocompatibility with human cells. Examples of various chemical approaches to surface-modify bacteria are presented. Chapter 2 systematically explores the immobilization of polystyrene nanoparticles on the surface of Escherichia coli (E. coli) and Salmonella typhimurium (Salmonella) cells. We compared the nanoparticle immobilization using four different conjugations strategies (electrostatic, covalent and ligand-ligand) and characterized the bio-hybrids systems using flow cytometry and confocal microscopy. Two different microscopy techniques were used: the first was based on 3D-reconstruction of z-stacks of single bacterium to investigate position of nanoparticles on the membrane; the second allowed for a large screening of bacteria followed by image processing determining the area of bacteria covered by nanoparticles. We observed that the efficiency of the surface modification is correlated to the initial nanoparticle/cell ratio and that non-covalent strategies allowed for increased number of biohybrids formed and nanoparticle attached. This study provides an overview of the variety of conjugation strategies (and their conjugation efficiencies) available for fabricating nanoparticle-decorated bacteria and presents us an opportunity to better understand and design bacteria-based target-specific drug delivery systems. Chapter 3 presents the characterization of the previously developed bacteria-nanoparticle biohybrids as potential carriers for nanoparticle transport. The viability of the various conjugates was determined by flow cytometry indicating a decrease in cell viability with increasing numbers of nanoparticles immobilized on the membrane. The motility was studied to investigate the impact of nanoparticle attachment on the bacterial velocity as well as the capacity to transport the cargo. Bacteria were able to transport nanoparticles when prepared with low ratios of nanoparticle/cell.

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Type
doctoral thesis
DOI
10.5075/epfl-thesis-8982
Author(s)
Bader, Lisa Patricia Elisabeth  
Advisors
Klok, Harm-Anton  
Jury

Prof. Esther Amstad (présidente) ; Prof. Harm-Anton Klok (directeur de thèse) ; Prof. Alexandre Persat, Prof. Inge Herrmann, Dr. Jordan Vacheron (rapporteurs)

Date Issued

2022

Publisher

EPFL

Publisher place

Lausanne

Public defense year

2022-07-08

Thesis number

8982

Total of pages

106

Subjects

Escherichia coli

•

Salmonella typhimurium

•

polystyrene nanoparticles

•

biohybrid microswimmers

•

surface-modification

•

in vitro characterization

•

motility

EPFL units
LP  
Faculty
STI  
School
IMX  
Doctoral School
EDMX  
Available on Infoscience
June 30, 2022
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/188795
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