Biology of Blood and Marrow Transplantation 12:87-91 (2006)

䊚 2006 American Society for Blood and Marrow Transplantation

1083-8791/06/1201-0116$32.00/0
doi:10.1016/j.bbmt.2005.10.004

Bionanotechnology Progress and Advances

Warren C. W. Chan

University of Toronto, Institute of Biomaterials & Biomedical Engineering, Terrence Donnelly Center for Cellular
and Biomolecular Research, Materials Science and Engineering, Toronto, Ontario, Canada

Correspondence and reprint requests: Warren C.W. Chan, PhD, University of Toronto, Institute of Biomaterials
& Biomedical Engineering, Terrence Donnelly Center for Cellular and Biomolecular Research, Materials Science
and Engineering, 160 College St., Toronto, ON, M5S 3E1, Canada (e-mail: warren.chan@utoronto.ca).

ABSTRACT
Advances in the nanotechnology research have provided a new set of research tools, materials, structures, and
systems for biological and medical research and applications. These nanotechnologies include the application
of fluorescent quantum dots for optical imaging, the design of metallic nanoparticle surfaces for ultrasensitive
biomolecular fingerprinting, and the use of nanostructures as hyperthermia agents for cancer therapy. Unlike
conventional technologies, unique properties can be incorporated into nanometer-size particles, structures,
and systems simply by changing their size, shape, and composition. Because of the tunable properties,
biologists and clinicians could custom-design a material for a specific research need. In this review article, we
highlight the recent advances and progress in Bionanotechnology research as well as provide future perspective
on this integrative field.
© 2006 American Society for Blood and Marrow Transplantation

KEY WORDS
Bionanotechnology ● Quantum dots ● Metallic nanostructures ● Multifunctional nanodevices

INTRODUCTION ing and therapeutics for treating cancer), and, because

of these demonstrations, new avenues and promises
Nanotechnology has rapidly emerged as an impor-
have been created for nanotechnology research.
tant field of research with potential effects in both
Terms such biomedical nanotechnology, bionanotechnol-
diagnostics and therapeutics. Nanotechnology is also
ogy, and nanomedicine are used to describe this hybrid
expected to accelerate fundamental biomedical re-
field.
search via the creation of novel state-of-the-art tools. There seems to be a natural fit between nanotech-
We loosely define nanotechnology research as the crea- nology and biology. With nanotechnology, a large set
tion, design, and manipulation of structures or parti- of materials with distinct properties (optical, electrical,
cles with dimensions smaller than 100 nm. Within this or magnetic) can be fabricated. Functionalities can be
size range, the structures or particles have properties added to nanomaterials by interfacing them with bio-
that can be tuned by changing the material’s dimen- logical molecules or structures (Figure 2). For exam-
sions [1,2] (Figure 1). For example, the fluorescence ple, magnetic nanoparticles with the conjugation of
emission of CdSe semiconductor particles is depen- the antibody Herceptin onto the surface can be used
dent on its size; a 2-nm particle emits blue light, as a contrast agent for tumor imaging (the antibody
whereas a 6-nm particle emits a red light. Nanotech- directs the magnetic nanoparticle to the tumor cells
nology research has traditionally been a major focus of while the magnetic nanoparticle provides a signal for
research by engineers and physical scientists because indicating detection) [3]. Diverse and large libraries of
nanomaterials were considered as important compo- biological molecules, such as antibodies and oligonu-
nents in the construction of electronic components cleotides, are available because they can be produced
and computer chips. The development of synthetic by using molecular biology or automated synthetic
protocols to reproducibly make nanomaterials and to techniques. Furthermore, the size of nanomaterials is
elucidate the relationship between their properties similar to that of most biological molecules and struc-
with respect to size and shape is a common area of tures; therefore, nanomaterials can be useful for both in
research. However, in the last 10 years, the biological vivo and in vitro biomedical research and applications.
and medical research communities have exploited the Thus far, the integration of nanomaterials with
unique properties of nanomaterials for various appli- biology has led to the development of diagnostic de-
cations (eg, contrast agents for cell and animal imag- vices, contrast agents, analytical tools, therapy, and

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problems associated with organic-based probes [9].

These nanometer-sized probes provide novel and unique
properties for biological applications that are not avail-
able with organic-based fluorophores. At the forefront of
this new class of inorganic probes are semiconductor
nanocrystals, also known as QDs. The physical proper-
ties and applications of QDs have been heavily investi-
gated in many physics and engineering laboratories
since the early 1980s [10-12]. QDs are defined as
particles with physical dimensions smaller than the
exciton Bohr radius; this gives rise to a unique phe-
nomenon known as quantum confinement. Quantum
confinement, which refers to the spatial confinement
of charge carriers (ie, electrons and holes) within a
material, embarks QDs with unique optical and elec-
tronic properties that are unavailable to semiconduc-
Figure 1. Rayleigh light-scattering of metallic nanostructures of
different sizes and shapes. This example shows the simplicity of
tors in bulk solids. Although initial interest in QDs
creating structures with unique properties, simply by changing the focused in physical applications (eg, making computer
structure’s dimensions. Adapted with permission [1] from the chips and light-emitting diodes), recent work by Nie,
American Association for the Advancement of Science, © 2001. Alivisatos, and their coworkers has highlighted the great
promise of QDs in biological applications [13,14]. As
biological probes, QDs are extremely bright (1 QD ⬇
drug-delivery vehicles. Mirkin et al. [4,5] exploited the
aggregation-based absorbance properties of gold nano-
structures for detecting genetic mutations; Kim et al.
[6] developed quantum dots (QDs) for sentinel lymph
node mapping; Han et al. [7] developed optical bar-
codes for the high-throughput analysis of proteins and
genes; and Hirsch et al. [8] developed metallic nano-
structures for treating cancer. Although these re-
searchers demonstrated some interesting applications
of nanomaterials, bionanotechnology research is still
in its infancy; few nanotechnology-based products are
in clinical use.
Visionaries in the field have predicted that one
day, researchers will incorporate multifunctionality
into nanomaterials. With these multifunctional nano-
devices, molecules and motors will guide nanomaterial
movements, sensors for diagnosis, actuators (which
are connected to the sensor) to release therapy, and a
secondary sensor to monitor the disease as it is being
treated. Such a device can diagnose, treat, and monitor
diseases such as cancer and Alzheimer disease. This
advancement will likely take at least 50 to 100 years to
develop. The foundation of nanotechnology is quickly
being put into place, and some applications of nano-
technology in biology have surfaced (although these Figure 2. Schematics of nanostructures integrated with biological
applications are nowhere near the complexity of the molecules and structures. A, Schematic of a semiconductor nano-
described multifunctional device). In this article, we structure called quantum dots (sphere) with a protein on its surface.
provide examples of current state-of-the-art bionano- This quantum dot/protein system can be used for labeling cells,
technology research. biosensing, or detecting tumors. Adapted with permission [10] from
the National Academy of Sciences, © 2004. B, Schematic of a nickel
nanostructure bound to an F1-adenosine triphosphatase biomolecular
SEMICONDUCTOR QDS FOR BIOIMAGING motor. The motor is a mechanical device to power up and direct the
nanostructure. One day, these motors may be important components
Recent developments in the field of nanotechnol- in building nanodevices for in vivo biomedical applications such as
ogy have led to the fabrication of a new generation of cancer treatment. Adapted with permission [20] from the American
inorganic nanostructures that overcome many of the Association for the Advancement of Science, © 2000.

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10-20 organic fluorophores), have high resistance to gonucleotide sequence on the gold, hybridization
photobleaching, have narrow spectral line widths, and occurs. With hybridization, the nanostructures are
have size- and materials-tunable emission that can be brought close together, and the solution color
excited by using a single wavelength. When QDs are changes (as an indicator of detection). This diagnos-
used in biological research and applications, these op- tic system has advanced toward a surface platform,
tical properties will lead to improved detection sensi- where silver staining is used for amplification of signal.
tivity for analysis and to simplification in experimental Metallic nanostructures can also be a platform for
and instrumental design. Because QDs have fluores- surface-enhanced Raman spectroscopy (SERS). Ra-
cence properties, they have been rapidly adapted into man spectroscopy is an analytical technique that mea-
many biological and medical research laboratories. sures the vibrational frequencies of chemical bonds
For in vitro applications, QDs conjugated to antibod- upon optical excitation. The major advantage of Ra-
ies and peptides are used for labeling receptors on man spectroscopy is that a tag or label is not required
fixed and live cells and tissues; QDs conjugated to for detection, but the technique is less sensitive than
oligonucleotides are used for genetic detection. For in fluorescence. However, the adsorption of molecules
vivo applications, successful demonstration of QDs as onto roughened metallic surfaces enhances the detec-
contrast agents for cancer imaging has already been tion capability of Raman spectroscopy. This technique
achieved. Kim et al. [6], Akerman et al. [15], and Gao is called SERS. Emory and Nie [19] showed that
et al. [16] have all shown the accumulation of QDs in SERS could detect a single molecule if that molecule
animal tumors. was adsorbed onto a metallic nanostructure with a
As QDs advance, there are several major avenues sensitivity comparable to that of fluorescence. SERS is
of research. The first is the continuing improvement starting to emerge as a viable analytical technique for
and development of QD optical probes with better clinical detection.
optical qualities (ie, high quantum efficiencies, narrow Therapeutic applications of metallic nanostruc-
spectral line widths, and more brightness). The sec- tures are also possibilities. Photothermal properties
ond is to understand the relationship between QDs can be engineered into the metallic nanostructures for
and biological systems (ie, elucidating how cells and laser ablation therapy. Metallic nanoshells, where a
animals uptake, process, and metabolize QDs). The nanometer-sized metallic layer is grown onto a glass
third is to study and understand the influence of bio- bead’s surface, and rods, where the nanoparticle’s as-
logical environments and conditions on the optical pect ratio is greater than 1:1, will produce heat when
and electronic properties of QDs. The fourth is to illuminated with a light source (that matches the
apply QDs toward clinical applications. nanostructure’s surface plasmon resonance). The sur-
face plasmon resonance is defined as a packet of elec-
trons on the surface of the metallic nanoparticle that
becomes excited after optical illumination. These me-
METALLIC NANOSTRUCTURES AS SENSORS
tallic structures can be used for hyperthermia therapy.
Many reports on the biological applications of We have highlighted some of the emerging appli-
metallic nanostructures as biological sensors have re- cations of metallic nanostructures. There are, however,
cently surfaced. Unlike QDs, metallic nanostructures many other applications (eg, molecular rulers and drug
were incorporated or used in many biomedical appli- delivery) for metallic nanostructures.
cations in the 1970s and 1980s [17,18]. Gold nano-
structures are used as contrast agents in electron mi-
croscopy or as detection probes for dipsticks (such as OTHER NANOSTRUCTURES
those for pregnancy tests). The manipulation of the
size, shape, and aggregation-dependent absorbance In addition to QDs and metallic nanostructures,
properties of metallic nanostructures and their dem- other nanostructures have been synthesized and used
onstrated applications have made this one of the most in biological and medical research. Among these
exciting areas of bionanotechnology research. nanostructures are carbon nanotubes, fullerenes, den-
The absorbance and scattering of light by metallic drimers, and magnetic nanostructures. Thus far, they
nanostructures is size and aggregation dependent have found applications in imaging (as contrast agents
[1]. A solution of gold nanostructures appears ruby or surface probe tips), drug delivery, and biosensing.
red, but the color of solution changes to blue when
the gold nanostructures are in close contact to one
another (or are aggregated). In this colorimetric TOWARD FUNCTIONAL DEVICES: INTEGRATION OF
diagnostic system, the biorecognition molecule oli- NANOSTRUCTURES WITH BIOLOGICAL MOLECULES
AND STRUCTURES
gonucleotide (which recognizes a gene for a specific
disease) is coated onto the surface of the gold nano- With recent advances in the synthesis of nano-
structure. When a gene sequence matches the oli- structures and a demonstration of their utility in bio-

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medical research, a major focus of the nanotechnology To add functionality into the nanostructures,
community is on the design of multifunctional nano- Soong et al. [20] coated the surface of inorganic nano-
devices. These devices will be a new type of advanced structures with biomolecular motors, such as F1-aden-
therapy for the treatment of cancer, Alzheimer dis- osine triphosphate synthase. This molecular motor
ease, or infections. Two major questions arise: (1) propelled the particles in solution. Hess et al. [21]
How can one organize nanostructures into a func- proposed the use of motor proteins with microtubule
tional device? and (2) How can one control the func- track systems to construct molecular conveyer belts to
tion of these nanostructures? Toward the engineering build nanoscale devices. They mimicked the biological
of these nanodevices, researchers use biology as an process of vesicle transport inside cells.
inspiration. Biological molecules and structures are
used for organizing nanostructure aggregations; bio-
logical motors are incorporated into the design to CONCLUSIONS
create rotors to control the nanostructure’s move- We have highlighted some of the recent develop-
ments. Currently, it is not clear how an engineer will ments in bionanotechnology research and provided
add sensing and actuating capability into the nano- some insights into the future of this hybrid field. As
structures. the field of bionanotechnology evolves, we envision a
The mimicking of biological systems for designing trend toward clinics, because nanotechnology will
nanodevices may be a powerful strategy. In biological likely allow a clinician to diagnose diseases at faster
systems, only 20 amino acids and 4 bases are needed to rates with improved sensitivity and specificity. The
coordinate the functions of thousands of proteins. The more futuristic goals of bionanotechnology, such as
amino acids and bases can assemble useful biological multifunctional nanodevices, will take much longer to
structures with noncovalent interactions. These inter- develop. Researchers are now starting to figure out
actions include hydrophobic-hydrophobic interactions, how to build such devices. This emerging field is
van der Waals forces, hydrogen bonding, and molecular exciting because of its possibilities.
stacking. Nanostructures, in general, are highly depen-
dent on molecular forces to assist them in maintaining
their monodispersity. Coordinating molecular forces, ACKNOWLEDGMENTS
which are used to fold proteins into a functional unit, W.C.W.C. would like to acknowledge Canadian
onto the surface of nanostructures is currently difficult Institute of Health Research (CIHR), Natural Sci-
because protein-folding mechanisms remain some- ences and Engineering Research Council (NSERC),
what unclear. A simpler and first step toward the use Canadian Foundation for Innovation (CFI), Ontario
of biology to mediate nanoparticle assembly is the Innovation Trust (OIT), the Connaugt Foundation,
use of biorecognition molecules. Biorecognition and Genome Canada for research support.
molecules can be coated onto a nanoparticle surface
to direct the formation of aggregates.
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