32 RESEARCH

Waqar Ahmed and St John Crean

DOI 10.1308/204268514X13859766312674

by Waqar Ahmed and St John Crean

Authors: Professor Waqar Ahmed*

and Professor St John Crean, School of
Medicine and Dentistry, University of
Central Lancashire, Preston PR1 2HE

* Corresponding author:
E: WAhmed4@uclan.ac.uk

Keywords: nanotechnology, tissue

regeneration, teeth, bioengineering

FACULTY DENTAL JOURNAL January 2014 • Volume 5 • Issue 1


Nature has always used nanotechnology to synthesise
molecular structures in the body such as enzymes, pro-
teins, carbohydrates and lipids, which form components
of cellular structures. However, the formal discovery of
nanotechnology is widely attributed to the American
physicist and Nobel Laureate Richard Phillips Feynman,1
who presented a paper called There’s plenty of room
at the bottom on the 29 December 1959 at the annual
meeting of the American Physical Society at California
Institute of Technology.
Feynman talked about the storage of information on a
very small scale, writing and reading in atoms, minia-
turisation of the computer, building tiny machines, tiny
factories, nanorobots and nanoelectronic circuits made
with atoms. He stated that: ‘In the year 2000, when they
look back at this age, they will wonder why it was not
until the year 1960 that anybody began seriously to move
in this direction.’ He did not specifically use the term
‘nanotechnology’. The first use of the word 'nanotech-
nology' was by Norio Taniguchi2 in a paper published in
1974 called On the basic concept of nanotechnology. K Eric
Drexler, an MIT graduate, later took Feynman’s concept
of a billion tiny factories and added the idea that they
could make more copies of themselves, via computer
control instead of control by a human operator, in his
1986 book Engines of Creation: The Coming Era of Nanotech-
nology, which popularised the potential of nanotechnol-
ogy.3 Fascination with nanotechnology has continued,
with features in films and news media.
Several definitions of nanotechnology have evolved.
The dictionary4 defines nanotechnology as ‘the art of
manipulating materials on an atomic or molecular scale,
especially to build microscopic devices’. The US govern-
ment5 states that:
Nanotechnology is the research and technology develop-
ment at the atomic, molecular or macromolecular level,
in the length scale of approximately 1–100nm range, to
provide a fundamental understanding of phenomena and
materials at the nanoscale and to create and use struc-
tures, devices and systems that have novel properties and
functions because of their small and/or intermediate size.
The Japanese6 have a more focused and succinct defini-
tion. ‘True nano’ is nanotechnology that is expected to
cause scientific or technological quantum jumps, or pro-
vide great industrial applications by using phenomena
and characteristics peculiar to the nano-level.
Regardless of the definition used, the properties of
matter are controlled at a scale between 1–100nm.
For example, chemical properties take advantage of
large surface-to-volume ratio for catalysis, interfacial
and surface chemistry for many applications. Mechani-
cal properties depend on the nanoscale for improved
strength, hardness, lightweight nanocomposites and
nanomaterials, altered bending and compression proper-
ties and nanomechanics. Optical properties involving the
nanoscale are improved absorption and fluorescence of
nanocrystals, single-photon phenomena and photonic
band-gap engineering. Fluidic properties give rise to
RESEARCH 33
enhanced flow using nanoparticles and adsorbed films.
Thermal properties improve the thermoelectric perfor-
mance in nanoscale materials and greater interfacial
thermal resistance.
Approaches to nanotechnology
Nanotechnology is a multidisciplinary field involving
chemistry, physics, biology, engineering, medicine,
dentistry, electronics and social sciences, which need to
be integrated together in order generate the next level
of development. Several approaches are used for mak-
ing nanostructures and materials and as the technology
develops further approaches may emerge.7 These have
generally been dictated by the technology available and
the background experience of the researchers involved.
Fuel cells, mechanically stronger materials, nanobiologi-
cal devices, molecular electronics, quantum devices,
carbon nanotubes and graphene have been made using
nanotechnology. Even social scientists are debating ethi-
cal issues in nanotechnology.
Two approaches are common. The ‘top-down’ approach
involves fabrication of device structures via monolithic
processing on the nanoscale and has been used with
spectacular success in the semiconductor devices used
in consumer electronics. The new smart phones, tablets
and other electronic devices provide powerful evidence
of success of his approach. The ‘bottom-up’ approach
involves the fabrication of device structures via system-
atic assembly of atoms, molecules or other basic units of
matter. This is the approach nature uses to repair cells,
tissues and organ systems in living things and indeed for
life processes such as protein synthesis. Tools are evolv-
ing that will give scientists more control over the synthe-
sis, characterisation and control of novel nanostructures,
which will yield a wide range of exciting new products for
healthcare in the near future – particularly in medicine
and dentistry.
Nanotechnology for tooth regeneration
Nanotechnology has enormous potential in the field of
dentistry.8,9 For example, tooth regeneration is an impor-
tant aspiration of the dental profession. The combina-
tion of tissue bioengineering along with the development
of genetically designed trigger nanopar¬ticles, which are
biomimetic with mineralised tissues, is beginning to bear
fruit in the manufacturing of in vitro tooth. Regeneration
of teeth can be divided into several specific areas:
• Regeneration or de novo formation of an entire, ana-
tomically correct tooth;
• Regeneration of the root;
• Regeneration of dental pulp;
• Regeneration of dentin that may either act as repara-
tive dentin to seal off an exposed pulp chamber or as a
replacement of current synthetic materials;
• Regeneration of cementum as a part of periodontium
regeneration or for loss of cementum and/or dentin
resulting from orthodontic tooth movement;
• Regeneration of periodontium including cementum,
periodontal ligament, and alveolar bone;
FACULTY DENTAL JOURNAL January 2014 • Volume 5 • Issue 1
34 RESEARCH
Waqar Ahmed and St John Crean
• Regeneration or synthesis of enamel-like structures
that may be used as biological substitute for enamel;
• Re-mineralisation of enamel and dentin.
For tooth regeneration, biomaterials have served primar-
ily as a scaffold for (1) transplanted stem cells and/or
(2) recruitment of endogenous stem cells. It is indispens-
able for the regeneration of tooth root, tooth crown,
dental pulp, or an entire tooth. Nanomaterials, which
mimic surface properties of natural tissues, are promis-
ing candidates for improving traditional dental tissues
engineering materials. The various forms of tooth-tissue
engineering related to nanotechnology and nanomateri-
als are described.
Biomimetic enamel regeneration
Enamel is the hardest material formed by vertebrates
and is the most highly mineralised skeletal tissue present
in the body. Mature enamel is composed of 95–97%
carbonated hydroxyapatite by weight, with less than 1%
organic material. Mature dental enamel has a complex
form, providing a striking example of a highly miner-
alised structure exquisitely adapted to absorb essential
mechanical and abrasive stresses throughout the lifetime
of the organism (Figure 1).
Figure 1 Scanning However, enamel cannot heal itself by a cellular repair
electron micrograph because enamel is both acellular and avascular. It loses
(SEM) of the surface
of an acid-etched
mineral substances owing to caries, trauma and erosion.
ground section of Restorations of damaged tooth tissues with artificial
mature mouse incisal materials represent the traditional therapeutic solutions.
dental enamel. Although many sophisticated materials are now available
Ordered arrays for restoration, their use is not yet completely satisfac-
of enamel prisms tory. A combination of tissue bioengineering and the de-
are constructed of
parallel bundles
velopment of genetically designed trigger nanoparticles,
of carbonated which are biomimetic with mineralised tissues, has begun
hydroxyapatite to bear fruit in the manufacturing of in vitro teeth tissue
enamel crystallites.10 – even the whole teeth. For example, the amelogenin
.................................... gene has been manipulated to adhere to hydroxyapatite
1 lesions and cavities are ongoing challenges in dentistry.
FACULTY DENTAL JOURNAL January 2014 • Volume 5 • Issue 1
nanoparticles. When these are directly shot to pluripo-
tential cells encapsulated in nanohydrogels they begin to
work on the formation of the enamel tissue.
Previous attempts to engineer enamel focused mainly
on chemical synthesis. Hydroxyapatite nanorod surface
with monolayers of surfactants synthesised and modi-
fied to create specific surface characteristics that allowed
the nanorods to self-assemble into an enamel prism-like
structure at a water/air interface. The size of the syn-
thetic hydroxyapatite nanorods can be controlled, and
synthesized nanorods were similar in size to both human
and rat enamel crystals.
The hydrothermal method has been used to synthesise
prism-like structures, consisting of fluorapatite (FA)
crystals similar to the dimensions of those seen in human
enamel. This method of nanotechnology has been widely
adopted to create nanorods, nanowires and whiskers,
and has already been shown to be an effective way of
creating different kinds of nanomaterials. However, the
majority of these synthesis methods were developed
using high temperature, high pressure and extremely
acidic pH, or in the presence of a concentrated solution
of surfactants. It is generally accepted that the biomi-
metic synthesis of enamel-like apatite structures under
physiological condition is an alternative restorative
pathway. Bio-inspired cooperative effects of an amino
acid (glutamic acid, Glu) and nano-apatite pariles can
result in the regeneration of enamel-like structure under
physiological conditions. Importantly, the mechanical
characteristics of the repaired enamel are well main-
tained by using this feasible enamel remodel. These
successful approaches of enamel regeneration imply a
potentially material-inspired strategy of nano assembling
in biomedical application and open the possibility that
in the future dental practice might drastically change,
al-lowing the manufacturing of teeth.
Enamel and dentine remineralisation
The prevention of tooth decay and the treatment of
Recently, biomimetic approaches have been used to
develop nanomaterials for the remineralisation of early
enamel lesions. Nano-hydroxyapatite (HA) is widely
studied as a biomimetic material for the reconstruction
of tooth enamel suffering from mineral loss and as an
effective anti-caries agent because of its unique poten-
tial for remineralisation. Nano-HA has the potential to
remineralise initial enamel caries lesions under dynamic
pH-cycling conditions. In addition, a concentration of
10% nano-hydroxyapatite may be optimal for reminerali-
sation of early enamel caries in vitro.
Nano-HA helped mineral deposition predominantly in
the outer layer of the lesion and only had a limited ca-
pacity to reduce lesion depth. Nevertheless, the reminer-
alisation effect of nano-HA increased significantly when
the pH was less than 7.0. There was a significant syner-
gistic effect of combined Galla chinensis extract (GCE)
and nano-HA treatment on promoting the remineralisa-
 RESEARCH 35

tion of initial enamel lesion. When GCE was added with

nano-HA, there was a significantly higher volume per
cent mineral present in the body of lesion. This would
not completely inhibit the deposition of nano-HA on
the outer layer of lesion in the remineralisation process,
therefore full remineralisation on the initial enamel le-
sion was obtained. SEM images showed that the crystals
on the surface layer in the GCE + nano-HA group were 2
arranged regularly and a densely uniform structure was
formed (Figure 2e), whereas irregularly arranged crystals
were present in the nano-HA group (Figure 2c).

The average size of the calcium phosphate crystals plays

an essential role in the formation of hard tissues, with a
significant influence on its intrinsic properties, including Figure 2 SEM images of the enamel surfaces in
solubility and biocompatibility. Evenly sized nano-apatite different groups (60 000×). Many micropores
particles (20 nm-sized HA and building blocks of biologi- and honeycomb structures were apparently on
cal apatite of dental enamel) could simultaneously repair enamel surface in DDW group (B). However,
and prevent initial erosive lesions in enamel compared after application of nano-HA, acicular crystals
had sedimented on the lesion surface and
with conventional HA crystals that are hundreds of nano- the cavities and microspores significantly
meters in length. In vitro studies also demonstrated that decreased. Meanwhile, the surface of the
nano-HA provides better remineralisation than micro- demineralised enamel appeared to be covered
HA. Analogues of nano-building blocks of biominerals by crystal, arranged in a thick and homogenous
should be highlighted in the entire subject of biominer- apatite layer (C). Some finger-like crystals
were distributed in a disorderly pattern on the
alisation.
enamel surface after being treated with GCE
and a honeycomb structure still remained in
Even though the remineralisation effect of nano-HA on some regions on the surface of lesion (D).
caries lesions is clear, the mechanism of action is still In the GCE + nano-HA group, the surface
debatable. Nano-HA promotes remineralisation through morphology was similar to that in the nano-HA
deposition onto etched enamel or by depositing apatite group. However, the crystals were arranged
regularly and a dense layer was also obtained
nanoparticles in the defects on demineralised enamel. after addition of GCE (E). Different-sized
Nano-HA may be acting to deliver a calcium source globules were formed on the lesion surface in
to the mouth, which can increase oral calcium levels, the NaF group (A).11
and has the potential to limit acid challenges by reduc- ................................................................................
ing enamel demineralisation while promoting enamel
remineralisation. The mechanism of remineralisation
involves HA acting as a calcium-phosphate reservoir,
helping to maintain a state of supersaturation with
respect to enamel minerals, thereby depressing enamel
demineralisation and enhancing remineralisation. This
is in accordance with the classic paradigm of ‘top-down’
ion-mediated crystalline growth to account for the
intricate biomineralisation strategies identified in nature.
Nano-HA, on the other hand, shows promising reminer-
alisation efficacy on enamel lesions in view of its unique
characteristics, including excellent deposition proper-
ties, which are in good agreement with the ‘bottom-up’
concept of particle-mediated nanoprecursor assembly
and mesocrystalline transformation in the biomineralisa-
tion process.

Dentine remineralisation is clinically significant for the

prevention and treatment of dentine caries, root caries,
and dentine hypersensitivity. However, dentine reminer-
alisation is more difficult than enamel remineralisation,
owing to the abundant presence of organic matrix in
dentine. An accepted notion is that dentine reminerali-
sation occurs neither by the spontaneous precipitation
nor by the nucleation of mineral on the organic matrix
(mainly type I collagen) but by the growth of residual
inorganic crystals in the lesions. Reconstitution and
remineralisation of dentine using nano-sized bioactive

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36 RESEARCH
Waqar Ahmed and St John Crean

Figure 3 Dental pulp tissue engineered for 35 days inside root canal using glass particles and betatricalcium phosphate was also
SHED cells (A) and natural dental pulp from premolar (B). It is possible tested in vitro; however, the mechanical properties of
to observe the formation of a healthy tissue without inflammatory signs original dentine could not be reproduced. Fortunately,
and a densification of odontoblast-like cell along dentin walls in the
SHED-originated tissue similar to the control. The engineered tissue
the biomimetic remineralisation scheme provides a
occupies the whole apical portion (C) and immunohistochemistry proof-of-concept for the adoption of nanotechnology
with proliferating cell nuclear activity (PCNA) and Factor VIII show a as an alternative strategy to remineralisation of dentine.
proliferative tissue with well-established and mature blood network (D Metastable amorphous calcium phosphate nanoprecur-
and E).12 sors were generated when polyacrylic acid was included
...................................................................................................................................................................................................................
in the phosphate-containing fluid. The nanoprecursors
were attracted to the acid-demineralised collagen matrix
and transformed into polyelectrolyte-stabilised apatite
nanocrystals that assembled along the microfibrils (in-
trabrillar remineralisation) and surface of the collagen
these proposals fibrils (interfibrillar remineralisation) to achieve dentine
remineralisation. The results revealed that guided tissue
may seemingly look remineralisation based on nanotechnology is potentially
useful in the remineralisation of acid-etched dentine
outrageous, but that is incompletely infiltrated by dentine adhesives and
partially demineralised caries-affected dentine.
inventions have always
been the brainchildren Dentin-pulp complex regeneration
Restorative dentistry is looking for techniques and
of outrageous ideas materials to regenerate the dentin-pulp complex in a
biological manner. This showed the great potential in
from the scientific the treatment of our most common oral health problem
and cavities. There is evidence suggesting that odonto-
community blasts (cells that produce dentin), dental pulp stem cells
(DPSC) and stem cells from human exfoliated decidu-
ous teeth (SHED) are able to produce pulp/dentin-like
tissues when seeded on specific condition or scaffolds. In
the process, advanced biomimetic scaffolding materials

FACULTY DENTAL JOURNAL January 2014 • Volume 5 • Issue 1

 RESEARCH 37

are versatile enough to provide a suitable 3D network to
accommodate these cells and guide their growth, organ-
isation and differentiation. One important step toward
regenerative endodontics was achieved when SHED
mixed with nanofiber peptide scaffold and injected into
full-length root canals were able to generate a dental
pulp. Figure 3 shows the presence of a pulp tissue fulfill-
ing the hollow passageway of the root canal, with prolif-
erative activity and blood network maturity comparable
to the ones observed in a young human dental pulp.
Another in vitro study showed that peptide-amphiphile
molecules provide a nanostructured, cell-responsive
matrix that is specifically conducive to dental stem cells.
The SHED and DPSC seeded in PA hydrogels show dif-
ference in morphology, proliferation, and differentiation
behaviour. SHED seem to be a suitable tool for soft-tissue
regeneration such as dental pulp, whereas DPSC might
be useful for engineering mineralised tissues like dentin.
Further development and successful application of these
strategies to regenerate dentin and dental pulp could
one day revolutionise the treatment of our most com-
mon oral health problem and cavities.
The future of nanodentistry
Biomedical scientists and clinicians all over the world are
working towards prevention and early delivery of care to
maintain human health. It is envisaged that nanotech-
nology will have a great impact in dental research and
improvement in current treatment methodologies lead-
ing to superior oral health care in the near future.
ing continuous plaque/calculus removal and metabolis-
ing trapped organic matter into harmless and odourless
vapour. These proposals may seemingly look outrageous,
but inventions have always been the brainchildren of
outrageous ideas from the scientific community. Predic-
tive tools like ‘lab-on-a-chip’ can utilise saliva as a media
to diagnose dental and other physical anomalies of the
human body.
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Nanomaterials will be used far more widely and will yield

superior properties and combined with biotechnology,
laser and digital guided surgery will thus provide excel-
lent dental care. Smarter preventive measures and earlier
interventions to avert craniofacial disorders using nano-
diagnostics seems a reality. Nanotechnology research will
definitely pave the way for development of tools, which
would allow clinicians to diagnose and treat oral malig-
nancies at their earliest stage.

Biomimetics and nanotechnology has given us the knowl-

edge to bioengineer lost teeth and remineralisation
of carious lesions. This is one field that has stimulated
immense interest among the dental and nanotechnology
researchers. Salivary glands can be a gateway to the body
for the delivery of precise molecular therapies using
nanoparticle-based drug delivery systems with fewer side
effects. Nanofillers have improved the aesthetic, physical
and mechanical properties of dental composite materi-
als.

Futuristic applications have been proposed on utilising

nanobots (nanoscale robots) to treat carious lesions,
dentin hypersensitivity, induce dental anaesthesia, teeth
repositioning (using orthodontic nanobots that could
directly manipulate periodontal tissues allowing rapid,
painless movement). Dentifrobots (nanorobots in denti-
frices) delivered through mouthwash or toothpaste could
patrol supra- and subgingival surfaces of tooth, perform-

FACULTY DENTAL JOURNAL January 2014 • Volume 5 • Issue 1