MMG 445 Basic Biotechnology eJournal

Practical Applications of Carbon Nanotubes in Medicine

Brian Martin
The past two decades have seen great technological advancements in the fields of optics, biochemistry, and physics, allowing the fundamentals of our own human biology to be understood
and controlled. At the forefront of this great understanding lies a tiny structure made of carbon
called a nanotube. At one one-thousandth the
width of a human hair, these promising nanostructures can be employed for various uses on
the cellular level, including the delivery of therapeutic molecules into cells. Many studies have
demonstrated that peptides, medicinal molecules,
and nucleic acids, when bonded to carbon nanotubes, are delivered considerably more safely
and effectively into cells than by traditional methods.
Addresses
Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI
Corresponding author: marti781@msu.edu
MMG445 Basic Biotechnology eJournal, 2006
This review comes from a themed issue based on
current advances in the fields of microbiology,
biotechnology, and pharmacology. It fulfills in part
the assignment of the contributing author in MMG
445, Basic Biotechnology, Department of
Microbiology and Molecular Genetics, Fall
semester, 2006.
Editors George M. Garrity and Terry L. Marsh
Board of Trustees, Michigan State University. All
rights reserved.

List of Abbreviations
CNT Carbon Nanotube
SWNT Single-Walled Nanotubes
MWNT Multi-Walled Nanotubes
FMDV Foot-and-Mouth Disease Virus
mAb Monoclonal Antibody
ELISA Enzyme-Linked Immunosorbent
Assay
AmB Amphotericin B
FITC Fluorescein Isothiocyanate

Introduction
In recent years, immense advancement has
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been made in the field of nanotechnology.

This emerging field will indefinitely become
a critical facet of many areas including
chemistry, biology, electronics, and optics,
and will provide unique opportunities for researchers to innovate in unimaginable ways.
Specifically, nanotechnology has nearly limitless potential in biomedical applications, as
our own biology is essentially a very complex system of nano-machines.
Thus emerges the field of nanomedicine,
which can be defined as the manipulation
and control of our own biological processes
through materials constructed on a nanoscale, approximately one one-thousandth the
width of a human hair. [1] This ability to reverse engineer our own biology allows us to
diagnose, cure, and prevent diseases with incredible accuracy not achievable through
conventional means. These treatments are
designed to be safer, more efficient, and
cause fewer side effects than traditional
medicine. [2] Specifically, CNTs, or carbon nanotubes, have been used to deliver
therapeutic molecules and micro-structures
to targeted cells and organs in a safe manner, which generates a low immunogenic response and is generally low in toxicity. [3]
This review will focus on recent developments in nanomedicine, specifically those
which utilize carbon nanotubes in the delivery of genes, medicinal compounds, and
peptides

ABCs of CNTs
Carbon nanotubes, originally discovered by
Iijima in 1991, are structures made up of
thin sheets of benzene ring carbons rolled up
into the shape of a seamless cylinder and are
often capped on at least one end by a spherical buckyball structure. These light, chemiMMG 445 Basic Biotechnology eJournal 2006 2:[pp pp]

Carbon nanotubes in medicine 27

cally stable cylinders conduct heat better

than diamond and are one of the strongest
materials in existence [4]. CNTs amazing
physical properties have astounded researchers from nearly every field of science,
providing them with a promising new tool
full of potential.
Two types of CNTs have been researched
for use in biomedical applications. The first
is a SWNT, or single-walled nanotube, consisting of a single sheet of carbon benzene
rings wrapped into the shape of a cylinder
[5]. The second is a MWNT, or multi-walled
nanotube, which is similar to a SWNT, but
contains multiple concentric layers of carbon sheets [6].
While pristine CNTs are insoluble in all solvents, there are several methods used to increase their biocompatibility. Strong acids
can be utilized to oxidize CNTs, improving
their solubility by adding carboxylic groups
to their structure. A more practical method is
to add functional groups such as ammonium
groups to the CNT walls and tips to increase
solubility [3].

CNTs in Drug Delivery

Drug delivery has been a major area of focus
for researchers aiming to improve the efficacy of therapeutic molecules. Some obstacles these researchers are trying to overcome
include poor drug distribution among cells,
unwanted damage to healthy tissue, toxicity,
and lack of the ability to select a particular
cell type for treatment [6]. However, researchers have found that CNTs possess the
characteristics to serve as a model drug delivery system, avoiding common barriers
typical of traditional drug delivery methods.
Many beneficial molecules can be bonded to
the walls and tips of these soluble CNTs, including peptides, nucleic acids, and various
drug molecules [7]. Important characteristics, such as the ability to easily cross cellular membranes, greatly enhance the potential
of CNTs for therapeutic uses.

SWNTs translocated easily into the cytoplasm or nucleus of a cell through its cell
membrane, without producing any toxic effects. Furthermore, the researchers were able
to covalently bond a peptide to the nanotube,
which was also easily absorbed. This peptide triggers the G protein function, which is
an important factor of signal transduction
[8].
As CNTs possess the ability to carry molecules of interest across the cytoplasmic
membrane and nuclear membrane without
producing a toxic effect, they prove to be a
very safe and effective drug delivery method
[8]. One example of this can be demonstrated by the delivery of the antibiotic amphotericin B, which is used in the treatment
of fungal infections. One major obstacle in
traditional delivery of this drug lies in the
fact that it has a low solubility and causes
membrane leakage in eukaryotic cells [9].
This obstacle has hindered the usefulness of
the drug when delivered through traditional
methods, such as encapsulation, due to the
low therapeutic index and the need for slow
delivery. Wu et. al postulated that delivery
of this antibiotic by means of CNTs would
reduce the amount of antibiotic necessary
resulting in improved potency reduced toxicity [9].
Using acid-oxidized MWNTs, the researchers covalently bonded AmB and the fluorescent marker FITC to uniformly distributed
wires on the carbon wall containing free
amino groups. They then studied the uptake
of these MWNT-AmB molecules into the
cell compared with an equal dosage of AmB
alone [9]. The results were exactly what
they had postulated, illustrating the immense
reduction in toxicity when AmB was delivered bonded to MWNTs in comparison with
AmB alone. When in the presence of AmB
alone at the highest levels tested, 40% of
cells died. Yet, with the same amount of
AmB attached to MWNTs, none of the cells
were reported dead, thus proving MWNTs
ability to reduce toxicity (Figure 1.) [9].

A recent study found that water soluble

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MMG 445 Basic Biotechnology eJournal 2006 2:[pp pp]

28 Martin

CNT, it can also be encapsulated inside of

the structure [11].

Figure 1. Percentage of dead cells in the

presence of MWNT-AmB and AmB. Cells in the
presence of AmB were significantly more likely to
die than those in the presence of AmB-MWNT.
From Wu et. al, 2005 Angewandte Chemie International Edition 44 (39): 6358-6362.

The researchers found that because the

nanotube-AmB complex can pass rapidly
through the cell membrane, it does not disrupt the membrane core the way that AmB
alone does [9].
Finally, the researchers tested the effectiveness of nanotube antibiotic delivery in antifungal activity against three species of
pathogenic fungi. The researchers tested this
using AmB bonded to MWNTs as well as
SWNTs, CNTs alone, and AmB alone. Surprisingly, the AmB that was bound to the
SWNTs and MWNTs were significantly
more effective as an antifungal agent than
was the AmB alone at equal dosage (Table
1.) [9]. This may be caused by increased
efficacy of the drug due to higher solubility.
Overall, this research demonstrates the value
of CNTs in drug delivery. Bound to CNTs,
molecules of therapeutic value can be administered safely and more effectively than
with previous delivery methods.

Gene Delivery by CNTs

Not only are nanotubes practical in the delivery of medicinal molecules, they can also
be utilized to deliver genes directly into the
cell and across the nuclear membrane. Gao
et. al found that not only can DNA molecules be attached to the tips and walls of a
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Currently, viral vectors for gene delivery are

in use and achieve high gene expression.
However, this method of delivery is far from
perfect, as viral vectors can be immunogenic
and can cause inflammation and oncogenic
effects [12]. Functionalized CNTs can
provide a safe nonviral vehicle for the delivery of DNA molecules into mammalian
cells, since these DNA-CNT structures are
produced under strict conditions in a cellfree manner [12].
In one study, Singh et. al tested the functionality of a SWNT-DNA complex as a
nonviral vector to deliver plasmid DNA into
an A549 cell. The team used the 7.2 kb
plasmid pCMV-Bgal in combination with a
single walled nanotube. This research concluded that the CNT-gene structure lead to
gene expression levels ten times higher than
that of naked DNA alone [12].
Not only was this complex very effective as
a vehicle for gene delivery, it was concluded
to be of low toxicity, soluble in aqueous solution, stable for long-term storage, and
highly modifiable for specific gene delivery
needs.

CNT Delivery of Peptides

Researchers from multiple universities in
France and Italy collaborated to test the immunogenic response of a peptide covalently
bonded to a SWNT, being the first to characterize such an interaction. Pantarotto et. al
performed this using a peptide from the
foot-and-mouth disease virus. This bonding
occurred on the SWNT wall at uniformly
distributed wires containing free amino
groups [10].
Pantarotto et. al tested the immunological
reactivity of the FMDV-NT structure and
free FMDV peptides with the specific mAb
21 x 27 using a Biacore3000 instrument,
which utilizes surface plasmon resonance
technology. This measures the increase inmass as an FMDV-NT or free FMDV pep
MMG 445 Basic Biotechnology eJournal 2006 2:[30 31]

Carbon nanotubes in medicine 29

Minimum inhibitory concentration (MIC) [micrograms mL]-1
CNT

C. parapsilosis

C. albicans

C. neoformans

AmB

20

>80

SWNT-NH3+

>80

>80

>80

MWNT-AmB

1.6

6.4

0.8

SWNT-AmB

1.6

13.8

0.8

Table 1. Antifungal activity of CNT-AmB conjugates. The antifungal activity of AmB, AmB bound to
CTNs, and CTNs alone against three species of pathogenic fungi. MIC refers to the minimum concentration
of each compound that displayed visible antifungal effects. The concentrations listed for both AmB-NT complexes refer to the amount of AmB in the complex. Both AmB-NT structures are significantly more effective
as an antifungal agent than was AmB alone in equal dosage. From Wu et. al, 2005 Angewandte Chemie International Edition 44 (39): 6358-6362.

tide solution passes over and reacts with a

fixed mAb. The Biacore 3000 test indicated
that the FMDV-nanotube structure reacted
in the same way with the antibody as the
free peptides. It also indicated that acetylated nanotubes alone did not generate a significant response from the mAb. A slower
association rate and increased response was
observed with the FMDV-NT in comparison
to the free FMDV peptides due to the increased molecular weight of the structure
(Figure 2.) [10].

mAb 21 x 27. From Hoebeke et. al, 2003 J. Am.

Chem. Soc. 125 (20): 6160-6164.

Similar tests using the ELISA method

agreed with the data from the Biacore3000.
Preliminary experiments by the same researchers demonstrated a strong humoral
immune response from mice immunized
with the FMDV peptideNT [10].
The results of this experiment signify a large
step in the utilization of CNTs for biomedical applications. Not only did researchers
find that binding molecules to carbon nanotubes increase the efficacy of the ELISA
technique, they also found that the CNTpeptide complexes prove to be incredibly
safe and effective in treating immunological
diseases. This novel method of vaccine delivery has the potential to improve or even
replace traditional methods.

Conclusion

Figure 2. FMDV-NT and Free Peptide Response Rates with mAb. The Biacore 3000 test
illustrates the response rate of FMDV-NT (bold
black line), free FMDV peptide (grey line), and
acetylated NT (thin black line) interaction with

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With the prospect of gene therapy, cancer

treatments, and innovative new answers for
life-threatening diseases on the horizon, the
science of nanomedicine has become an
ever-growing field that has an incredible
ability to bypass barriers previously thought
unavoidable. The properties and characteristics of CNTs are still being researched heavily and scientists have barely begun to tap
the potential of these structures. Single and
multiple-walled carbon nanotubes have alMMG 445 Basic Biotechnology eJournal 2006 2:[pp pp]

30 Martin

ready proven to serve as safer and more effective alternatives to previous drug delivery
methods. They can pass through membranes, carrying therapeutic drugs, vaccines,
and nucleic acids deep into the cell to targets
previously unreachable. They serve as ideal
non-toxic vehicles which, in some cases, increase the solubility of the drug attached, resulting in greater efficacy and safety. Overall, recent studies regarding CNTs have
shown a very promising glimpse of what lies
ahead in the future of medicine.

Acknowledgements
I would like to thank Dr. George Garrity and Dr. Terry
Marsh for providing me with the opportunity to
enhance my skills as a scientific writer and assimilate
myself into the modern community of scientific
research. I would also like to thank my reviewers for
their time and constructive criticism.

5.

6.

Bianco A, Kostarelos K, Partidos C,

Prato M: Biomedical applications of
functionalized carbon nanotubes.
Chemical Communications December
2004, 571-577.
This article reviewed experiments done by
the authors themselves, covering a broad
but useful range of topics. I enjoyed reading
this article during my research.
7.

Lin Y, Taylor S, Huaping L, et al: Advances toward bioapplications of

carbon nanotubes. Journal of Materials
Chemistry January 2004.
This review article goes in depth on the applications of nanotubes, while giving a comprehensive look at the growing field of
nanomedicine and biosensors. Very interesting background information.
8.

References and recommended

reading
Papers of special significance that have been
published within the period of review are highlighted as follows:
of significance
of special significance
1.

Kralj M, Pavelic K: Medicine on a small

scale. EMBO Reports 4 2003, 11: 10081012.

2.

Leary S, Liu C, Apuzzo M: Toward the

emergence of nanoneurosurgery:
part III-nanomedicine: targeted nanotherapy, nanosurgery, and progress
toward the realization of nanoneurosurgery. Neurosurgery June 2006,
58(6): 1009-1026.
This article provides a very good basis for
the basic understanding of targeted nanotherapy, including the targeting of specific
cells by nano-structures containing medicines.
Bianco A, Kostarelos K, Prato M: Applications of carbon nanotubes in
drug delivery. Current Opinion in
Chemical Biology December 2005, 9(6):
674-679.
I found useful visuals as well as important
explanations in this article that gave me a
good grip on my topic. This article is very
thorough.

Dresselhaus M, Dresselhaus G, Eklund

P, Saito R: Carbon nanotubes. Physics
World January 1998.

Pantarotto D, Briand JP, Prato M, Bianco A: Translocation of bioactive

peptides across cell membranes by
carbon nanotubes. Chemical Communications 2004, 16-17.

9.

Wu W, Wieckowski S, Klumpp C, et al:

Targeted delivery of amphotericin B
to cells by using functionalized carbon nanotubes. Angewandte Chemie
International Edition September 2005,
44(39): 6358-6362.
This was an extremely useful primary article
on the use of CNTs in delivering the antifungal agent AmB. I was astonished to see the
results of this experiment.
10. Hoebeke J, Graff R, Briand JP, et al:
Synthesis, structural characterization, and immunological properties of
carbon nanotubules functionalized
with peptides. J. Am. Chem. Soc.
2003, 125(20): 6160-6164.
This primary article was invaluable in the understanding of solubility in nanotubes. It also
provided excellent support for my topic in detailing the process and benefits of attaching
peptides to CNTs.

3.

4.

Jamieson V: Carbon nanotubes roll on.

Physics World June 2000.

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11. Gao H, Kong Y, Cui D: Spontaneous

insertion of DNA oligonucleotides
into carbon nanotubes. Nano Letters
2003, 3(4): 471-473.
12. Singh R et. al: Binding and condensation of plasmid DNA onto functionalized carbon nanotubes: toward the
construction of nanotube-based gene
delivery vectors. J. Am. Chem. Soc.
2005, 127 (12): 4388-4396.
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