Article 1 Prussian blue Nanoparticles and its Analogues as New-Generation T1-Weighted

MRI Contrast Agent for Cellular Imaging

Abstract

There are insufficient achievements in the field of cancer diagnosis and treatment for new dual agents,
which would provide health care specialists the ability to simultaneously image patients’ cancerous
tissues as well as treat the diseases. Prussian blue (ferric hexacyanoferrate) is a nontoxic FDA approved
compound used clinically as an antidote for thallium and radioactive cesium poisoning. In this thesis
development of simple methods for the synthesis of biocompatible Prussian blue nanoparticles (PBNPs)
and its analogues as well as their applications for magnetic resonance imaging (MRI) contrast agents and
drug delivery have been studied. The extensive magnetic properties investigations show that Prussian
blue nanoparticles and gadolinium doped analogue nanoparticles significantly shorten the T1 relaxation
time in aqueous solution and in HeLa cells treated with PBNPs, demonstrating their potential use as MRI
contrast agents. Although the relativity values of Prussian blue nanoparticles are approximately an order
of magnitude lower than the typical commercial Gd3+-based T1 contrast agents but it is found to be
comparable to the values obtained for the MnO nanoparticles-based T1 agents. In order to provide high
contrast, gadolinium doped Prussian blue nanoparticles (Gd-PBNPs) were prepared. It was also found that
the Gd-PBNPs can shorten the T1 relaxation time significantly and provide potential use for clinical
applications. In order for Prussian blue and its analogues nanoparticles to be concurrently utilized as drug
delivery agents they must be biocompatible and capable of crossing the plasma membrane. Therefore,
Prussian blue nanoparticles and related analogues were synthesized and functionalized by carboxylic
acids such as citric acid as capping agents to control size distribution. To study the intracellular uptake of
Prussian blue and analogue nanoparticles, their surfaces were functionalized separately with the small
molecule dyes such as 5-carboxyfluorescein and Alexa Fluor® 350 cadaverine, as well as the anticancer
agent. Confocal fluorescence imaging of HeLa cells treated with the functionalized nanoparticles shows
fluorescent signals in the cells suggesting intracellular uptake of the Prussian blue and Gd-PB
nanoparticles. The HeLa cells internalized Prussian blue nanoparticles and gadolinium-containing
Prussian blue nanoparticles could also enhance the T1 MRI contrast. The results clearly show that these
nanoparticles can be used as an effective T1 contrast agent for cellular imaging. Functionalized Prussian
blue nanoparticles and related analogues with both MRI contrast and drug delivery capabilities may
become powerful dual agents for simultaneous cancer treatment and assessment of treatment
effectiveness.

Reference:

Shokouhimehr, Mohammadreza. Prussian blue nanoparticles and its analogues as new-

generation T1-weighted MRI contrast agents for cellular imaging. Diss. Kent State University,
2010.
Article 2 Platinum-Monolayer Shell on AuNi0.5Fe Nanoparticle Core Electro catalyst with
High Activity and Stability for the Oxygen Reduction Reaction

Abstract

We describe a simple method for preparing multimetallic nanoparticles by in situ decomposition of the
corresponding Prussian blue analogue, which is adsorbed on carbon black. The example involves the
AuNi0.5Fe core of the PtML/Au1Ni0.5Fe core-shell electro catalyst for the oxygen reduction reaction.
The core contains 3-5 surface atomic layers of Au, which play an essential role in determining the activity
and stability of the catalyst.ThePtML/AuNi0.5FeelectrocatalystexhibitedPtmassand specific activities of
1.38 A/mgPt and 1.12 × 10-3 A/cm2Pt, respectively, both of which are several times higher than those of
commercial Pt/C catalysts. Its all-noble-metal mass activity (0.18 A/mgPt, Au) is higher than or
comparable to those of commercial samples. Stability tests showed an insignificant loss in activity after
15 000 triangular-potential cycles. We ascribe the high activity and stability of the PtML/AuNi0.5Fe
electro catalyst to its hierarchical structural properties, the Pt-core interaction, and the high
electrochemical stability of the gold shell that precludes exposure to the electrolyte of the relatively active
inner-core materials.

Reference:

Gong, Kuanping, Dong Su, and Radoslav R. Adzic. "Platinum-monolayer shell on AuNi0. 5Fe
nanoparticle core electro catalyst with high activity and stability for the oxygen reduction
reaction." Journal of the American Chemical Society 132.41 (2010): 14364-14366.

Article 3 Electrically Triggered Release of a Small Molecule Drug from a Polyelectrolyte

Multilayer Coating

Abstract

Electrically triggered drug delivery represents an attractive option for actively and remotely controlling
the release of a therapeutic from an implantable device (e.g., a “pharmacy-on-a-chip”).Here we report the
fabrication of Nano scale thin films that can release precise quantities of a small molecule drug in
response to application of a small, anodic electric potential of at least þ0.5 V versus Ag/AgCl. Films
containing negatively charged Prussian Blue (PB) nanoparticles and positively charged gentamicin, a
small hydrophilic antibiotic, were fabricated using layer-by-layer (LbL) assembly. When oxidized, the PB
nanoparticles shift from negatively charged to neutral, inducing dissolution of the film. Films with
thicknesses in the range 100-500 nm corresponding to drug loadings of 1-4 μg/cm2 were characterized.
We demonstrate control over the drug dosage by tuning the film thickness as well as the magnitude of the
applied voltage. Drug release kinetics ranging from triggered burst release to on/off, or pulsatile release,
were achieved by applying different electric potential profiles .Finally, the in vitro efficacy of the released
drug was confirmed against Staphylococcus aureus bacteria. Given the versatility of an external electrical
stimulus and the abilityoftheLbLassemblytoconformallycoatavarietyofsubstratesregardlessofsize, shape,
or chemical composition, we maintain that electrically controlled release of a drug from an LbL-coated
surface could have applications in both implantable medical devices and transdermal drug delivery
systems.
Reference

Schmidt, Daniel J., Joshua S. Moskowitz, and Paula T. Hammond. "Electrically triggered release of a
small molecule drug from a polyelectrolyte multilayer coating." Chemistry of Materials 22.23 (2010):
6416-6425.

Article 4 In situ Controllable Growth of Prussian Blue Nanocubes on Reduced Graphene

Oxide: Facile Synthesis and Their Application as Enhanced Nanoelectrocatalyst for H2O2
Reduction

Abstract

As a single-atom thick carbon material with high surface area and conductivity, graphene provides an
ideal platform for designing composite nanomaterials for high-performance electrocatalytic or
electrochemical devices. Herein, we demonstrated a facile strategy for controllably growing high-quality
Prussian blue Nanocubes on the surface of reduced grapheneoxide(PBNCs/rGO), which represents a new
type of graphene/transition metal complex heterostructure. The merit of this method is that the composite
nanomaterials could be produced directly from GO in an in situ wet-chemical reaction,where the
reduction of GO and the deposition of PBNCs occurred simultaneously. The obtained composite
nanomaterials were characterized by transmission electron microscopy (TEM), Xray photoelectron
spectroscopy(XPS),X-raydiffraction(XRD),thermogravimetricanalysis(TGA),Ramanspectroscopy,and
electrochemical techniques. It was found that uniform PBNCs with controlled size and good dispersion
were directly grown on the surface of graphene nanosheets. Moreover, we also investigated the
performance of PBNCs/rGO nanocomposites as amperometric sensor toward reduction of H2O2. Such a
sensor showed a rapid and highly sensitive response to H2O2 with a low detection limit (45 nM), which
might find promising applications in developing a new type of enzyme less biosensor.

Reference

Cao, Linyuan, et al. "In situ controllable growth of Prussian blue nanocubes on reduced graphene oxide:
facile synthesis and their application as enhanced nanoelectrocatalyst for H2O2 reduction." ACS applied
materials & interfaces 2.8 (2010): 2339-2346.

Article 5 Amperometric immunosensor based on multiwalled carbon nanotubes/Prussian

blue/nanogold-modified electrode for determination of α-fetoprotein

Abstract

In this article, a conspicuously simple and highly sensitive amperometric immunosensor

based on the sequential electrodeposition of Prussian blue (PB) and gold nanoparticles
(GNPs) on multiwalled carbon nanotube (MWCNT)-modified glassy carbon electrode (GCE)
surface is proposed for the detection of α-fetoprotein (AFP). By comparison with PB, the
MWCNT/PB composite film had been proven to show much better electrochemical stability
and a larger response current. The electrodeposited GNP film can be used not only to immobilize
biomolecules but also to avoid the leakage of PB and to prevent shedding MWCNT/PB composite film
from electrode surface. The performance and factors influencing the performance of the immunosensor
were investigated. Under optimal experimental conditions, the proposed immunosensor for AFP was
observed with an ultralow limit of detection (LOD) equal to 3 pg/ml (at 3δ), and the linear working range
spanned the concentrations of AFP from 0.01 to 300 ng/ml. Moreover, the immunosensor, as well as a
commercially available kit, was examined for use in the determination of AFP in real human serum
specimens. More significant, the assay mentioned here is simpler than the traditional enzyme-linked
immunosorbent assay (ELISA), and an excellent correlation of levels of AFP measured was obtained,
indicating that the developed immunoassay could be a promising alternative approach for detection of
AFP and other tumor markers in the clinical diagnosis.

Reference:

Jiang, Wen, et al. "Amperometric immunosensor based on multiwalled carbon nanotubes/Prussian

blue/nanogold-modified electrode for determination of α-fetoprotein." Analytical biochemistry 407.1
(2010): 65-71.