Nazmun Nahar PHA 6146 April 09, 2023

Engineering gold nanoparticles for molecular diagnostics and biosensing

Abstract:

In the field of research, nanoparticles as well as nanomedicines have become significant

inventions in the field of science and technology in the last few years. In biomedical research,

researchers are most commonly using AuNPs, which are both unbitten and secured nanoparticles

developed over the past 20 years. Meanwhile, researchers are experiencing some challenges in

the way of delivering drugs to specific sites, inadequate clinical trials for nanoparticles, efficient

manufacturing processes for commercial production and also some other major problems

researchers are facing. This gold nanoparticle is capable of either absorbing or scattering visible

and ultraviolet light into the electromagnetic field and can be used as a biosensor in the visible

and ultraviolet regions of the electromagnetic field. Develop a mechanism that metal component

combined with inorganic or organic, protein, lipid and polymer these are recognized elements for

detection strategies based on molecular complementarity and antigen-antibody affinity, that

gives sensing platforms [Naresh & Lee & Stater et al.,]. Moreover, it has some iconic

physicochemical properties, such as its shape, size, solubility, stability, and low toxicity, which

make it an excellent tool for detection and diagnosis. Researchers use AuNPs as "small magic

balls" in biomedical research. As an example, AuNPs are synthesized and amalgamated into

antibodies, proteins, and biomolecules. Additionally, AuNPs release Au+ ions that lyse cancer

cells and prevent DNA replication, thus destroying them. NPs have proved effective during the

COVID-19 pandemic in order to facilitate diagnosis as well as early detection of SARA-CoV-2,

therapeutics, imaging and vaccines against Coronavirus during this pandemic situation.

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Background:

The modern technology marvel of nanotechnology. Nanotechnology is the use of materials in

molecular or atomic level, usually 1nm to 100 nm. Due to its compact size and shape, surface

area-to-volume ratio, molecular charge, conductivity, strength, molecular weight, technologies

and therapeutic methods can be created using

nanotechnology particular, chemotherapy for

cancer uses it in the diagnosis, treatment, and

prevention of disorders. Nanomaterials can be

classified based on organic, inorganic, carbon

based and composite [Naresh & Lee, 2021; Figure-1: a model of Gold nanoparticles. Freitas De
Freitas, L., Varca, G., Dos Santos Batista, J., &
Stater et al., 2021]. There are some toxicity Benévolo Lugão, A. (2018).

issues associated with nanoparticles and need

more careful use with them. Some instances of the use of nanotechnology in medicine include

Drug delivery to specific cells or tissues using nanoparticles, which can improve treatment

efficacy while reducing adverse effects by the no exposure of healthy cells.

Manufactured gold nanoparticles is used for biosensing and molecular diagnostics [Ferreira et

al., 2020], A number of techniques, such as chemical synthesis, laser ablation, or electrochemical

deposition, can be used to create engineered gold nanoparticles. It is possible to generate

nanoparticles in certain sizes and shapes, which can then be functionalized with biological

molecules to provide particular biosensors for a range of analytes. There are numerous ways to

functionalize gold nanoparticles with biological molecules, including physical adsorption,

covalent interaction, and self-assembly.

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Nanotechnology can be utilized to make nano scaffolds that can be used to produce new tissues

in organs where there has been an injury. Eventually, it can be created to only target cancer cells,

preserving healthy cells while precisely delivering medication or radiation to the tumor site. In

general, nanotechnology has the power to transform medical care by enabling earlier and more

precise diagnosis and more effective patient treatments.

Gold nanoparticles are used in photothermal therapy (PTT), which uses light, usually in the near-

infrared (NIR) region to create heat that kills cancer cells. Because gold nanoparticles absorb

light at particular wavelengths, they can be utilized to heat up and kill cancer cells. This method

is being investigated as a potential cancer treatment for many cancer types.

AuNP’s ability to interact with bacterial cell membranes and alter their structure and function is

the main factor contributing to their antibacterial effectiveness. When AuNPs engage with the

phospholipid bilayer inside the bacterial cell membrane, it damages the membrane and kills the

bacterium and stops DNA replication. The generation of reactive oxygen species (ROS) such as

superoxide ions and hydrogen peroxide by AuNPs is another way that they exhibit antibacterial

action. Bacterial cell death can result from oxidative damage caused by these ROS to the cell

membrane and intracellular components.

AuNPs can be used during the pandemic to treat COVID-19 by preventing viral replication.

AuNPs can bind with the spike proteins on the SARS-CoV-2 virus's surface and stop them from

binding to the receptors on host cells, preventing the virus from entering the host cells. AuNPs

can also retire the viral RNA polymerase, which is necessary for viral replication.

In conclusion, due to Nanotechnology, researchers can produce nanomaterials for the treatment

of complicated diseases like cancer using different methods. By this technology, it is now

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possible to detect cancer in its early stage and start treatment. Still, there are some drawbacks of

this new invention, and researchers are working to overcome these issues.

Literature review

1. Biological applications of gold nanorods

Gold nanoparticles are important materials for sensing, photothermal treatment, and imaging

because they have optical characteristics that can be changed by altering their form, size, or

environment. Because of their large surface-to-volume ratio and sensitivity to their surroundings'

dielectric properties, gold nanoparticles exhibit distinctive optical characteristics. Shifts in the

colorimetric and extinction spectrum may be seen as a result of these features, which can be

influenced by variations in dielectric constants. Since they scatter visible light and have

associated plasmons, gold nanoparticles are utilized as sensors.

Electrochemical, photochemical, and electrochemical processes are used to create gold nanorods.

By sticking mostly to the side sides of the nanoparticle, CTAB promotes rod production. Gold

nanoparticles are now practical materials for advances in biology and medicine, although

cytotoxicity is still a problem. By

covering them with PEG, gold nanorods

have been discovered to lessen the

cytotoxic effects on human leukemia

cells. This is probably because to

desorption from their surface or free

CTAB from inadequately filtered

Figure-2: Schematic outlining the bioconjugation of gold
nanorods to antibodies via amide bond formation between free
primary amines on the nanorod surface and carboxylic acids
present on the antibodies.

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solutions. It is believed that receptor- mediated endocytosis will obscure the effects of gold

nanoparticles.

Due to the absorbing and scattering characteristics, gold nanorods already have drawn more

interest as instruments for biological sensing, imaging, drug delivery and therapies. Development

of gold nanorods coated with Au 2 S/AuAgS for tracking refractive index changes due to target

binding [Huang et al.,]. Hyper-Rayleigh scattering spectroscopy was utilized to measure HIV-1

DNA with great sensitivity and selectivity utilizing gold nanorods [Darbha et al.,]. A possible

method to find rare or highly infectious microbes is to employ single-molecule detection using

gold nanorod sensors and F1-ATPase motors.

Gold nanorods have attracted interest as drug delivery systems, although DNA release is caused

by photoinduced morphological changes in the nanorods. For remote gene expression regulation,

Chen et al. Attached are gold nanorods to the thiolated gene of an enhanced green fluorescent

protein. Gold nanorods have uses in nanomedicine, nanobiotechnology, drug release,

photothermal therapy, and optical switches to regulate gene expression. Compared to traditional

chemotherapies, photothermal treatment is less damaging to human tissues.

2. Nanomedicine for Cancer Immunotherapy:

Using Gold Nanoparticles and CT Imaging to Monitor Cancer-Specific T-Cells in Vivo." During

course of the investigation, mice models of breast and melanoma cancer were utilized.

After injecting mice with AuNPs that were coated in cancer-specific peptides, CT imaging was

used to monitor the distribution of the particles. According to the findings, gold nanoparticles

never aggregated when they were found in tissue, but they did so when they were found in

malignant tumors. After being isolated, the mouse T-cells were likewise treated with the identical

gold nanoparticles that had been used on the other cells. These tagged T-cells were injected into

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the mice in the form of a solution. Using CT imaging, the researchers were able to track the

tagged T-cells as they moved closer and closer to the tumors they were studying. They found that

the tagged T-cells aggregated in the tumors and had the ability to target the cancer cells precisely;

this demonstrates the efficacy of cancer

immunotherapy.

Results Description
Gold nanoparticles Accumulated in tumors but not in healthy
tissues
Labeled T-cells Accumulated in tumors and targeted cancer
cells
Optimal imaging time 24 hours after injection provided the best
visualization of labeled T-cells

Gold nanoparticles toxicity The optimal time for tagged T-cell

visualization was 24 hours following
injection.

The researchers also conducted a study to determine the optimal time to photograph the tagged

T-cells after the injection of the substance. They found that imaging was most successful 24

hours following the injection since this allowed for the clearest picture of the tagged T-cells that

were present within the tumors. This discovery was made. The possible toxicity of the metal

nanoparticles was also investigated by the researchers, who found that the mice were unaffected

in any way by their exposure to the metal nanoparticles.

Overall, the results of the study demonstrated that gold nanoparticles have the potential to serve

as an effective tool for monitoring the progress of cancer immunotherapy by making it possible

to track cancer-specific T-cells in vivo. This approach offers the possibility of improving the

results of cancer treatments by providing data in real-time regarding the distribution and

effectiveness of T-cells that are particular to the disease being treated [Meir et all., 2015]

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3.Gold nanoclusters with enhanced tunable fluorescence as bioimaging probes

Nanotechnology provides a variety of functional nanoparticles appropriate for imaging and

diagnostic probes, such as fluorescent gold clusters (FGCs), which exhibit stable and controllable

visible emission, a small hydrodynamic diameter, good biological compatibility, and are

predicted to revolutionize medical diagnostic tools. The absorption of surface plasmons by FGCs

varies with size and shape from visible to NIR, while the absorption of surface plasmons by gold

nanoparticles varies with size

and shape from visible to NIR.

FGCs were synthesized

utilizing a strong reducing

agent based on borohydride, a

capping agent based on thiol,

etching of gold nanoparticles

with suitable ligands, and

reduction of gold salt with a

weaker reducing agent in the Figure-3: (a) Most successful schemes toward functional
fluorescent gold clusters suitable for bioimaging application. (b)
Time line of the first report of different approaches for synthesis
presence of a strapping agent. of fluorescent gold clusters.

The most successful FGC synthesis methods include thiol-based reduction and stabilizing,

protein-based reduction and stabilizing, phosphate salt reduction and stabilization, as well as

thiolated stabilization. Protein-capped fluorescent gold clusters (FGCs) are most promising for

bioimaging probes, such as BSA capped red emitting FGCs, folate-functionalized FGCs,

Herceptin conjugated BSA capped FGCs, gadolinium functionalized BSA capped FGCs, insulin
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capped red fluorescent FGCs, human transferrin stabilized FGCs, and graphene oxide-based

composites with FGCs.

Gold nanoclusters (FGCs) are used as fluorescent probes, peptide stabilized FGCs, biological

labels, and for in vivo tumor imaging. Signal transduction is necessary for both the detection of

important biomarkers and the capacity to view where these biomarkers are distributed throughout

the body [Ferreira et al., 2020]. According to recent research, FGCs can effectively reduce

background signal because of their large two-photon absorption cross sections. FGCs-based

imaging probes are effective at giving meaningful information while being less hazardous than

other fluorescent nanoprobes. These compounds can be synthesized with a compact

hydrodynamic diameter, red or NIR emission, controllable visible emission, powerful emission,

functionalization, and imaging using a conventional fluorescence microscope. A deeper

understanding of this process requires further research

4. Gold nanoparticle-mediated photothermal therapy: applications and opportunities for

multimodal cancer treatment

Nanoparticle-mediated Photothermal Treatment (PTT) selectively hyperthermias tumor tissue

with laser light. Due to their easy gold-thiol conjugation, superior optical characteristics, and

passive tumor cell accommodation, gold-based nanoparticles (AuNPs) are the leading

therapeutic platform. AuNP-mediated Photothermal Treatment in conjunction with therapies,

may treat metastatic lesions and malignant cells beyond the irradiation region. PEG-coated

AuNPs are the only PTT AuNPs in clinical studies and have outstanding clinical safety.

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Gold nanoparticles (AuNPs) with chemotherapy may enhance cancer treatment by boosting drug

infiltration, cellular uptake, and protein

expression on cancer cell membranes.

Based on laser intensity and temperature,

researchers have hypothesized two

processes for light-induced oligonucleotide

release from AuNP surfaces. PTT releases

oligonucleotides from AuNP surfaces and

endosomes, improving gene control. PTT

and immunotherapy prevent tumor recurrence and metastasis.

AuNPs are optical contrast agents in optical coherence tomography, and photoacoustic imaging

for image-guided PTT. Photoacoustic imaging can monitor tissue thermal dosage during PTT,

allowing researchers to tune irradiation settings to limit cell death. PA imaging detects lymph

node micrometastases, whereas MRTI works Figure-4: (a) The heat generated by AuNPs in
response to NIR light (b) PTT can lead to either
cellular necrosis or apoptosis.
better on static tissues. AuNP-mediated PTT

may enhance imaging and therapy outcomes. Based on laser power and temperature, it may

release conjugated or encapsulated compounds at sick locations, chemo-sensitize cancer cells,

and immunotherapy, and modify cellular apoptosis or necrosis. The authors emphasize the

importance of conducting additional research and development work in order to fully understand

the potential of metal nanoparticles in the treatment of cancer. [Xu et al., 2022]

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5. Nanoparticle-based imaging of inflammatory bowel disease

Chronic irritable bowel disease (IBD) can affect the GI system. Nanotechnology and imaging

techniques can diagnose IBD early, monitor disease activity, and track medication results at the

cellular and molecular levels. This article discusses nano-imaging and IBD, and IBD develops

from inflammation and immune cell infiltration. Previous research suggests a connection

between incorrect immune responses, environmental variables, and the microbiota in the gut;

nevertheless, the specific cause of inflammatory bowel disease is still unknown [Hamidi N, et

al.,]

IBD imaging uses PET/CT, PET/MRI, PET/SPECT, CT enterography, MRE, and SPECT.

CT is recommended for

its inexpensive, rapid

scan time, and get

available. CT requires a

Barium-sulfate contrast

materials for GI tract

and IBD detection.

MRE and MREC are

rarely used for IBD due

to extended scan times, Figure-5: Representative images of inflammatory bowel disease (IBD) imaging as a
function of imaging modality.

and more expensive. IBD

sites are full of inflammatory cells.

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Molecular imaging scaffolds lipid-based nanoparticles (NPs) are tiny and pass through the

intestinal wall and are attached to the vector target. Lipid-based NPs can directly conjugate the

targeted moiety with CT/MR metal or SECT/PET radionuclides. Dendrimers may be heavy.

Nuclear medicine molecularly images cancer, but IBD is ignored. Philips PET/CT increases PET

sensitivity by 10. In preclinical studies, PEG-gold NP-targeted CT imaging agents target

intravascular inflammation and cancer. Gold NPs attenuate CT signals better than commercial

iodinated contrast agents.

Gold nanoparticles (NPs) may measure IBD gut wall inflammation. Spectral CT, dual energy,

and k-edge subtraction enable CT signal deconvolution. Inflammation-targeted MRI agents

include Gd, Mn, SPIO, and Fe NPs. The first FDA-approved liver iron oxide NP, AMI-25 or

Feridex, is biocompatible and low-toxic.

6.DNA-tailored plasmonic nanoparticles for biosensing applications

Metal nanoparticles (PNPs) have plasmonic characteristics , resulted in it can be biosensor

probes and optical nanomaterial building blocks. AuNPs or AgNPs interact with various

biological molecules via electrostatic binding as well as through functional groups such as thiols

and amines. DNA has become a potential nanostructure template [Watson-Crick et al.,]. PNPs

offer intriguing optical features but cannot recognize biomolecular targets for detection.

Bioplasmonic hybrids from DNA-modified PNPs may amplify signals, recognize targets, and

assemble highly organized nanostructures. First created oligonucleotide-modified AuNPs for

plasmonic color-change detection of biomolecules [Mirkin and colleagues et ai.,]. Au-S

chemistry is used to modify AuNPs with thiolated DNA. Label-free colorimetric sensing

strategies have been developed without the need to modify DNA to Au. Studies on SERS for

detecting biomolecules and enhancing signals have been carried out. Mirkin, Dong, Yu, and

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Moskovits created SERS sensors for DNA, protein, and adenosine utilizing AuNPs, stained

Silver and DNA labeled with a Raman dye. Based on a dye-labeled DNA target, Nam and

colleagues developed a high-yield synthesis technique for nanodumbbell structures that are

SERS-active core-shell structures composed of gold and silver. For more reliable biosensing

applications, fluorescence should be addressed. The Gedded researcher group devised a rapid

and sensitive microarray and MEF-based DNA detection technique using low-power

microwaves, whereas Lakowicz group used MEF-active surface-bound silver NPs. SPR imaging

biosensors can analyze chemical interactions in real time, but tiny fluctuations in reflection

efficiency trend limit them. Colloidal AuNPs can be employed for multiplexed parallel analysis

of high-throughput samples and SPR imaging biosensor constraints. Keating and colleagues

boosted DNA detection to 1 fM using NP-enhanced SPR imaging. Corn Researchers group used

enzyme-modified surfaces as well as nanoparticles coated with DNA in order to enhance SPR

imaging using NPs. Optimized conjugation procedures can build nanostructured probes from

NPs and biomolecules. DNA is being used to construct new nanostructures, including plasmonic

nanostructures, which increase signals without amplification cycles. The lack of high-yield

synthetic processes prevents the widespread use of these materials.

7. Semiconducting polymer nanoparticles for amplified photoacoustic imaging:

Semiconducting

polymer nanoparticles

(SPNs) are novel

photonic materials

with biological

applications. They

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recently improved photoacoustic (PA) imaging sensitivity and resolution. The PA imaging

technique combines optical excitation and ultrasonic detection to penetrate deep into tissues. PA

imaging relies on contrast agents since their absorption and photothermal conversion dictate PA

signal sensitivities. Melanin and lipids are PA imaging endogenous contrast agents (ECAs)

[Jiang & Pu, 2017; Yao & Wang, 2011 et al.,]. These compounds have low aqueous solubility,

photothermal stability, photostability, biosafety, and phototoxicity. SPNs are used for

fluorescence imaging, biological analysis, in vivo afterglow imaging, and distribution imaging.

Hence, high-sensitivity and specificity exogenous imaging agents for PA imaging are needed.

SPNs are used as PA imaging nanoplatforms in this study.

Photothermal conversion and photostabilities make SPNs adaptable nanoplatforms. By rationally

designing their -conjugated backbones, SPNs may be made to satisfy specific imaging needs.

Nevertheless, Semiconducting polymer nanoparticles (SPNs) have been created as versatile

nanotheranostic platforms for tumor PA imaging and PTT. PTT has performed when PA signal

has strongest and had the best tumor therapeutic impact. SPNs show potential in PA imaging, but

they must be addressed before clinical use. Biodegradable amphiphilic polymers can bundle SPs

to create SPNs.

8.Smart dual-mode fluorescent gold nanoparticle agents

Biomedical investigations use fluorophore-mediated molecular sensing, and gold nanoparticles

(GNPs) are best for biomarker-specific, highly sensitive sensing and imaging. Discussing GNP

fluorescence alteration approaches. mediated molecular sensing and imaging in biomedical

research. Discussing GNP fluorescence alteration approaches. CT, MRI, PET, ultrasound, and

optical imaging give additional in-vivo information. Fluorophore-mediated optical biosensing

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with biomarker targets is cheap, sensitive, and straightforward to utilize. GNPs are excellent for

biosensing since they are chemically inert and nontoxic.

The surroundings, frequency of incoming light, GNP, and fluorophore characteristics define a

nanoparticle's EM field's fluorescence emission intensity. GNP form, size, and distance from a

fluorophore affect fluorescence. Quantifying the optical link between the GNP and fluorophore

and employing plasmon field strength mathematical models can achieve this. Larger GNPs decay

faster and have a greater plasmon field beyond the particle surface. Radiative decay, absorption,

and intrinsic quantum yield influence a fluorophore's plasmon field-affected quantum yield.

This improves biosensing and bioimaging specificity depending on the fluorophore and GNP;

inserting fluorophores near a GNP may restore fluorescence. Spacers should be straight and

hydrophilic to preserve GNP-fluorophore spacing. The fluorophore/GNP & Silver combination

may be engineered to quench and intensify fluorescence, creating a more sensitive molecular

beacon [Jeong H. Kim et al.,]. GNP contrast agents may detect biomarkers in biofluids, cell

cultures, and animal models.

Figure-7: Theoretical estimation of fluorescence output (Φ) of various fluorophores, affected by a 10, 30, or 50 nm
GNP, with respect to the distance from the GNP surface. The black, horizontal dashed line at Φ =1 shows the
fluorescence level without GNP.

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Hydrophilic GNP surfaces prevent biological adsorption. Immobilizing fluorophore-conjugated

spacers and short polymers together improves distance control, while changing the solution's

ionic strength prevents aggregation. High GNP concentration alters inter-GNP fluorescence.

Fluorophore-mediated molecular imaging and sensing using GNPs increases specificity and

sensitivity. Theoretical estimate and experimental optimization may help produce effective

optical contrast agents.

Conclusion & Future Direction

Throughout this literature review, different studies have been summarized to show that AuNPs

have the ability to diagnose and biosensing of different diseases, especially cancer cells. The

gold nanoparticles have different physicochemical properties especially size range 1 nm to 100

nm, unique optical properties, and surface plasma resonance biocompatibility and

biodegradability for accurate diagnostics and biosensing and target drug delivery. Additionally,

gold nanoparticles have some limitations associated with adverse reactions. In the further

proposal, I will try to focus on playing with the size of my formulation to reduce toxicity

associated with gold nanoparticles.

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