Functionalised Gold Nanoparticles For

Controlling Pathogenic Bacteria

Diagnosis And Treatment Of Bacterial Infections

Contents

Introduction
Antibiotic Drug Resistance
Gold nanoparticles (GNPs)
Diagnosis of bacteria using the color change phenomenon
Different steps in the use of GNPs
Other diagnosis techniques
Destruction of bacteria using GNPs delivering chemical
payloads
Enhancement of photodynamic therapies using GNPs
Hyperthermic destruction of bacteria using GNPs
Combination of photothermal and chemical effects for
pathogen destruction
Conclusion

Introduction
Pathogenic bacteriaarebacteriathat can causeinfection.
Antibioticsare a type
ofanti-microbial
used specifically againstbacteria.

Antibiotics revolutionized medicine

in
the 20th century and have together
with vaccination lead to the near
eradication of diseases such as
tuberculosisin the developed world

Www.Tumblr.com/BacteriaC

Antibiotic Drug Resistance

Their effectiveness and easy access led to overuse, especially
in live-stock raising, prompting bacteria to develop resistance.
This
has
led
to
widespread
problems
withantimicrobialandantibiotic resistance, so much as to
prompt theWorld Health Organizationto classify antimicrobial
resistance as a "serious threat.
Bacterial tolerance is not just due to resistance, but also to
the formation of persistent cells that have gone into a
dormant state where they are no longer sensitive to
antibiotics. On the molecular level, this process is controlled
by a number of advancedcytotoxinsproduced by the
bacteria themselves in order to survive.

Antibiotic Drug Resistance

http://evolution.berkeley.edu

The increasing number of bacterial

strains that are resistant to available
pharmaceutical compounds is a vital
issue for public health.

Innovative
approaches
will
be
required to improve the methods for
both diagnosis and destruction of
these organisms.

Emerging infectious diseases and the

increase in incidence of drug
resistance
among
pathogenic
bacteria have made the search for
new antimicrobials inevitable.

Here, we consider the possible role that

can be played by technologies based on
gold nanoparticles.

Vancomycin Resistance

Gold nanoparticles (GNPs)

Nanoparticles usually ranging in dimension from 1-100 nanometers (nm)

have properties unique from their bulk equivalent.
With the decrease in the dimensions of the materials to the atomic level,
their properties change. The nanoparticles possess unique physico-chemical,
optical and biological properties which can be manipulated suitably for
desired applications .

The nanoparticles are finding important applications in the field of medicine.

The small size and the high surface to volume ratio i.e., large surface area of
the nanoparticles enhances their interaction with the microbes to carry out a
broad range of probable antimicrobial activities.

Au particles are particularly and extensively exploited in

organisms because of their biocompatibility.
Gold nanoparticles (Au) generally are considered to be biologically
inert but can be engineered to possess chemical or photothermal
functionality.
GNPs undergo a localized surface plasmon resonance, or electron
oscillation, with light, which can be exploited to generate a color
change or localized heating, amongst other possibilities.

Diagnosis of bacteria using the colorchange phenomenon

Nanoparticle-based methods have the potential to be rapid,
easy to use, and inexpensive, while at the same time,
maintaining a high level of accuracy.
Tiny Gold Particles can easily slip through Cell membranes,
making them good candidates to deliver drugs directly to
target cells.
Nanoparticles enter cells by taking advantage of a route
normally used in vesicle-vesicle fusion, a crucial process
that allows signal transmission between neurons.
The detection of bacterial DNA by probes based on GNPs
originated in the pioneering work of Mirkin et al.,who
demonstrated that as little as 10 fmol of an analyte
oligonucleotide could be detected by exploiting the
agglomeration phenomenon of GNPs.

 Suspensions of isolated gold nanospheres

have a strong localized surface plasmon
resonance with green light.
To a lesser extent, they also absorb blue light
as a result of interband electronic transitions
in the gold.
A GNP suspension will thus appear red to the
eye, which corresponds to the wavelengths
of any light that is not absorbed.
Now, far less red light is transmitted and the
resulting color is blue or purple. This change
in optical properties can be exploited for
both diagnostic and therapeutic purposes.

Different steps in the use of

GNPs
GNPs must be first nucleated from a suitable precursor under tightly
controlled conditions to give the desired particle size and shape.
Afterwards, GNPS typically are functionalized (i.e. coated) with one or more
molecules that will provide desired chemical properties, such as molecular
recognition or suppression of an immune system response.
GNP targeting to a desired cell or location. Attachment of an antibody that
is specific for the desired target.
At their target location, GNPs are activated to perform their function;
shown here is their illumination with a laser.
The result of GNP activation is verified, as performed here by using a
yellow fluorescent dye (propidium iodide) that is indicative of cell death.

Other diagnosis techniques

Other transduction modalities, besides color change, are also possible. GNPs have been used
to improve the sensitivity of quartz crystal microbalances (QCMs), or various modes of
fluorescence detection.
Wang et al. has used a QCM functionalized with GNPs for the detection of DNA from a
particular E. coli strain.
Two sizes of GNPs were used to increase the sensitivity.
First, GNPs of 18 nm diameter were immobilized onto the QCM surface and served as a
platform to support ssDNA that bound specifically to the biotinylated DNA of the target
bacteria presenting in the analyte liquid.
The use of this gold layer allowed a greater number of ssDNA molecules to be bound to the
QCM, thus increasing the sensitivity of the sensor.
Once the biotinylated DNA from the target organism had bound to the sensor,
avidinfunctionalized GNPs of 70 nm diameter were added in order to further amplify the signal.

This scheme allowed the detection of bacteria without requiring an enrichment of the sample.

Destruction of bacteria
using GNPs delivering
chemical payloads
Usage of Nanoparticles in drug delivery is enhanced due to the
stability, selectivity or functionality of the pharmaceutical
molecule.
Surfaces of GNPs are particularly suitable to serve as a stable and
non-toxic platform, on which pharmaceutical compounds can be
attached to and subsequently deployed for medical treatment.
Vancomycin conjugated to GNPs has increased its efficacy more
than 50-fold to destroy vancomycin-resistant enterococci
(Enterococcus faecium and Enterococcus faecalis).
Gram-positive bacteria : The underlying mechanism involves the
binding of vancomycin to the D-Ala-D-Ala moieties of the peptide
unit that is present on the bacterial wall of the which leads to
inhibition of the cellular processes of the organism.
Gram-negative bacteria : Mechanism involve physical attachment
of the conjugates to the bacterium and usually double the
required dose.

The vancomycin molecules on

each GNP(red barrels) bind to
the glycosides (hexagons) on
the surface of a vancomycin
resistant bacterium via the AlaD-Ala moieties of the peptide
unit.
The net effect of vancomycincoated GNPs is to bring a large
number of vancomycin
molecules into close proximity
to the organism, from which
they can disrupt its function.
The ability to bind to and/or
penetrate the cell wall and, in
doing so they are able to
deliver a large number of
antibiotic molecules into a
highly localized volume.

Enhancement of
photodynamic therapies
using GNPs
Photodynamic therapy (PDT) refers to the use of light activated
agents to destroy cancer cells or pathogenic organisms.
Dye molecules (the photosensitizer) interact with light and
with the normal ambient triplet oxygen molecules to produce
highly reactive singlet oxygen, which in turn can be used to
destroy target organisms by oxidation.
There have been attempts to target the photosensitizer
molecules to the site of pathogen infection using an antibody
specific to the pathogen to avoid systemic phototoxicity.
GNPs have been considered as stabilizers for antimicrobial
photosensitizers (toluidine blue or methylene blue) which is
found to be stable in air, water and PBS resulting in a four-fold
increase in the bactericidal effect in S. aureus and E. coli.

Hyperthermic destruction of
bacteria using GNPs

GNPs can be used to deliver a localized hyperthermic effect

because of their plasmon resonance.
Active targeting of GNPs involve attaching molecules that
specifically bind to the surface of the target organism.
Laser irradiation can then be directed to the localized region of
the infection and at a specific frequency and power to provide
both minimum damage to the intervening tissue and
maximum excitation of the plasmon resonance.
Usage of either continuous or pulse-mode radiation will
generates heat (photothermal effect) at the site of the GNPs
resulting in hyperthermic destruction of bacteria.
E.g. GNPs were conjugated to monoclonal anti-protein A, which
binds to the peptidoglycan section of the bacterial wall. When
strong laser light was applied to in vitro suspensions of
bacteria in the presence of the conjugate, death of the
bacteria was observed.

Combination of photothermal and

chemical effects for pathogen
destruction

1.
2.
3.
.

GNPs are functionalized to achieve more than one purpose,

basically chemical payloads and hyperthermia-inducing
properties.
Here vancomycin is conjugated with polygonal GNPs which
increases the efficacy of vancomycin in three ways:
it improves the delivery of the drug payload
its improves binding to the pathogen
it confers an ability to bind to Gram-negative organisms
Once the conjugates are bound to their target, photothermal lysis
of the bacteria can be induced using the plasmon resonance
phenomenon.
E.g. lysozymes that were loaded into GNP microcapsules irradiated
by light, these capsules heat up and burst, thus releasing their
payload which then destroyed a suspension of Microccus
lysodeikticus.

Conclusion

GNPs provide a versatile platform for a range of emerging

antibacterial technologies, in which the unique optical
properties, chemical stability, and ability of GNPs to be
conjugated to a wide variety of molecules can be exploited.

GNPs can be used to improve the sensitivity and speed of

techniques to detect or diagnose bacteria.
GNPs have been employed both as stabilizing and
photothermal agents in the form of conjugates with
antimicrobial agents and antibodies resulting in improved
antimicrobial activities into the target cells.
Targeted plasmon-induced hypothermia also can be
achieved using GNPs.
Hybrid combination strategies in which plasmonic heating is
used to liberate a chemical payload, constitute a further
development of these concepts.