DEPARTMENT OF PHYSICS AND NANOTECHNOLOGY

SRM INSTITUTE OF SCIENCE AND TECHNOLOGY

18NTO408T- INDUSTRIAL NANOTECHNOLOGY

Nanoparticles in bone substitutes and dentistry

18PYB101J Module-IV Lecture-9 1

Tissue engineering and regenerative medicine (TERM) aims to conjugate
engineering and biological properties to create functional substitutes for damaged
and diseased tissues.

The strategy used on TERM research combines three essential elements - scaffolds,
stem cells, and growth factors - to produce a tissue engineered construct.

Scaffolds provide the support for cell growth and tissue

formation. For that, they are seeded with stem cells.

Growth factors are also included as they regulate the

differentiation and proliferation processes.

Bone tissue regeneration is one of the greatest

challenges for TERM. The anatomical complexity of
bone, allied with the high mechanical stress to which it
is exposed, makes it unique, and almost impossible to
replicate.

Nevertheless, some strategies have been exploited,

with positive results.
18PYB101J Module-IV Lecture-9 2
Nanotechnology has made it possible to create structures within the same size as
those that constitute naturally occurring bone, opening a new era for TERM.

Hence, nanoparticles (NPs) can be used to modify scaffolds properties, leading to

enhanced characteristics such as superior mechanical properties and
osteointegration, osteoconduction, and osteoinduction.

Moreover, NPs can be applied to deliver drugs in a controlled and dependent manner,
either systemically or locally.

In another approach, NPs can be used to label cells, namely stem cells, enabling the
continuous cell tracking and monitoring of its fate.

Antibodies, labeling probes, hydrophobic or hydrophilic molecules, DNA, and/or

igonucleotides are some of the molecules that can be linked to NPs, allowing a
tailored application for the desired purpose.

18PYB101J Module-IV Lecture-9 3

Liposomes.

Liposomes are vesicles composed of one or more concentric bilayers, of natural

nontoxic phospholipids.

Each layer surrounds an aqueous compartment, where the head polar groups of
phospholipids are in contact with aqueous phases while the hydrophobic parts are
packed together.

liposomes as “simply the vesicles which form spontaneously when isolated natural
cell membrane phospholipids are shaken in water.

Taking advantage of this characteristic, the author used the liposomes as model
for the cell membrane, but nowadays these particles are also used in a wide range
of applications including drug delivery and imaging.

Liposomes properties are affected by several

parameters, such as lipid composition and
organization, surface charge, preparation method, and
size.

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The preparation methods affect the selfassembly of phospholipids, leading to
different types of liposomes, which can be classified by their size and lamellarity.

it is necessary to incorporate the desired molecules inside the liposome structure.

If the molecule is hydrophobic, it is mixed with an organic solvent, and it will be

integrated inside the hydrophobic part.

But when the cargo is hydrophilic, it must be added as an aqueous solution, being
retained in liposome inner part.

Specifically, liposomes within the nanoscale range can be administrated along with
drugs, or with other types of NPs.

A drawback of these strategies is a fast recognition of liposomes by the

reticuloendothelial system, which speeds up the process of their elimination from
the bloodstream.

This phenomenon can be reduced by using stealth liposomes, that is, liposomes
coated with hydrophilic biocompatible polymers, such as poly(ethylene glycol)
(PEG).
Bioceramics and Bioactive Glass NPs.
Bioceramics, glasses, and glass–ceramics are characterized as materials with an
inorganic/nonmetallic composition that are biocompatible and applicable for
biomedical uses.

From a chemical perspective, these materials can be divided into two classes:
(i) calcium phosphate groups, that includes hydroxyapatite and b-tricalcium
phosphate; and
(ii) others, such as alumina, bioactive glass systems, and bioactive glass–ceramic
systems.

Bioceramic NPs such as hydroxyapatite (HAp), calcium defective HAp (CDHA), and
tricalcium phosphate have been combined with natural and synthetic polymers
forming nanocomposite materials.

Bioactive glasses are amorphous silicate-based materials that can form a chemical
bond with bone tissue.

The silicon component of this material is released when the bioactive glass is
implanted, inducing the formation of a calcium deficient hydroxyapatite layer on its
surface.

This layer interacts with collagen fibrils of damaged bone, allowing bioactive glass to
bond to the surrounding tissue. 6
Hydroxyapatite NPs

This is the main composition of mineralized tissues of the human body

(Ca10 (PO4)6·2(OH)). It is a natural calcium phosphate ceramic, predominant in 97%
enamel.

Teeth are acellular in nature, thus it cannot be logically repaired like a bone. Thus
regenerating the enamel surface is a significant challenge.

The nano sized HAp particles can easily integrate into the dental tubules.

HAp helps to reduce dental hypersensitivity.

HAp NPs can bind strongly with proteins as well as with bacterial and plaque
fragments.

Their high biological activity and reactivity enable them to bind to the dentin apatite
and tooth enamel.

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 Hydroxyl apatite nanoparticles can fit well with the very small cavities present in
the enamel originated by acidic erosion.

The HAp NPs are adsorbed robustly to the enamel of the teeth and thus retard
auxiliary erosive demineralization.

Various toothpaste, mouth-rinsing solutions integrate these nanocrystals to repair

the enamel surfaces.

The biomimetic function of hydroxyapatite is to protect the teeth by making a film

of artificial enamel around the tooth.

The granular hydroxyapatite is employed in

dental clinical rehearsal to reform
periodontal shortcomings.

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Zirconia NPs

The use Zirconia (Zirconium dioxide, ZrO2) has considerable significance in dental
science.

It has similar metallic properties and color like tooth. Zirconia is a chemical oxide
which is insoluble in water. Thus, it reduces the bacterial adhesion and has low
cytotoxicity.

Zirconia implants encompass glorious resistance against corrosion and carry, as

well as sensible biocompatibility.

Moreover, high fracture resistance can be acquired by ZrO2 because of energy

retention property throughout the conversion of polygonally shaped molecules into
monoclinic ones.

Zirconia NPs is a bio inert material, the encapsulation by animal tissue is weak and
also the unleash of remains virtually unnoticeable.

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Additionally, Zirconia is osseo conductive, thereby it facilitates bone formation.

Nano zirconia-alumina materials combine the physical and chemical properties of

ceramic material.

In these NPs, low percentage of tetragonal ZrO2 particles is in an aluminum oxide

matrix. Thus, the toughness and longevity which are the principal interest in the
dentistry are retained.

Alumina/zirconia nano composites are

new implant materials which show
better efficacy as compared to the
ceramic materials.

Zirconia oxide nanoparticles are

found to have anti-biofilm activity
against certain bacteria and therefore
they can be effectively used as a
polishing agent in dental practices.

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Silica

In the field of dentistry silica NPs used as dental filler. various dental filler products
developed to improve their mechanical properties.

Tooth polishing is a conventional practice, which uses silica particles. Silica

particles are used in polishing for their biocompatibility and low cost

Polishing of teeth surfaces is often done to protect the enamel surfaces.

Thus, polishing prevents dental caries, which acts as a primary defense

mechanism against the cariogenic bacteria.

Modified silica nanoparticles are used to treat dental hypersensitivity. Enamel loss
exposes dentinal tubules, thereby increasing the risk of dental hypersensitivity.

Over the years, a number of desensitizing agents are commercially available that
aim at occluding dentinal tubules.

Unfortunately, the products can penetrate only up to a small depth into the dentinal
tubules which may not combat the daily adverse conditions.

mesoporous silicas have been widely researched over the past two decades.
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Silver NPs

AgNPs have also been studied for use in several areas of dentistry which includes
endodontics, dental restorative material, dental prosthetics, dental implants.

Incorporation of AgNPs decreases microbial colonization over dental parts and

increases oral health.

As the nanoparticles possess small size having the larger surface area, they show
the antimicrobial effect at very low level.

Because of its minute size, AgNPs can able to penetrate easily the bacterial cell
membrane resulting in rapid bactericidal activity.

Silver can interfere with DNA and proteins by interacting with —SH groups, and
also alters the base pairing, DNA unwinding, cell wall synthesis and respiratory
processes, resulting in bacterial death

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Scaffolds

The scaffold is an essential part in TE approaches where its key function is to

provide a temporary mechanical support in the defect site and a suitable
environment for the cells to grow.

bone is among the hardest load-bearing tissues in the human body with a significant
Young’s modulus (E = 17.5–18.9 GPa.

The major goal of scaffold-based strategies is to obtain an implant to replace the

native tissue while supporting the regeneration process without further
complications.

• porosity for cell migration;

• balance between surface hydrophilicity and hydrophobicity for cell attachment;
• mechanical properties comparable to natural tissue to withstand natural loading
conditions;
• degradation capability so that it gets completely reabsorbed after implantation;
• nontoxic byproducts;
• 3D matrix.

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From a TE perspective, bone is a complex nanocomposite, hierarchically organized,
with an organic extracellular matrix, strengthened by inorganic calcium phosphate
NPs, HAp crystals.

As polymers are easily modified and functionalized, they are commonly used as the
organic portion, mostly as nanofibers.

This configuration is beneficial for bone tissue formation because it mimics the type
I collagen fibers found in natural bone.

However, their compatibility to bone tissue and their mechanical strength is often
inferior when compared to calcium phosphate ceramics.

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Adding nanomaterials as a filler to these matrix polymers can augment the
mechanical properties of the scaffolds.

As a result, different nanomaterials are being studied as scaffold fillers to investigate

their effect on cells.

ceramic nanomaterials such as HA and bioactive glass, carbon-based materials like

CNTs and graphene, and metallic based NPs such as gold and titanium oxide (TiO2)
have been widely investigated

In addition to their role in enhancing the mechanical properties of scaffolds, it has

also been suggested that nanoscale materials can have specific influences on cell
signaling pathways.

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CNTs
Incorporation of multiwall carbon nanotubes (MWCNT) and HANP in a collagen
matrix in freeze dried scaffolds showed improved mechanical properties, cell
spreading and proliferation response.

scaffolds containing 0.5% MWCNT were examined in vivo in a rat calvarial bone
defect model for healing 8mm diameter defects.

After 12 weeks of healing, newly formed bone and connective tissue was observed,
which were more abundant when compared with collagen-HA and control groups.

Metallic NPs incorporated in scaffolds are also being developed for bone TE
applications. however, the challenge is to find biocompatible particles.

Gold (Au), silver (Ag) and TiO2 have long been known as interesting candidates
in the field of TE.

Silver nanoparticles (AgNP) have been incorporated in scaffolds for enhancing

osteogenic performance and Gold NPs (AuNPs) showed exceptional potential in
enhancing cell differentiation.

For primary osteoblasts, it has been shown that a diameter of 20 nm exhibits the
highest osteogenic effect, while for the human adipose-derived stem cells, 30–50 nm
particles performed the best. 16
Incorporation of calcium phosphate NPs, with similar size and functionality as the
nanosized inorganic ceramic particles that are naturally present in bone, is a way
to improve scaffolds properties.

This strategy often leads to an increased osteocompatibility and mechanical

performance of such nanocomposites over purely polymeric bone substitutes

HAp NPs were doped with gadolinium and then used within a polycaprolactone
(PCL) nanofibrous scaffold. HAp acquired a dual purpose, both improving bone
conductivity and acting as a magnetic resonance contrastagent.

The incorporation of 30–40 nm sized HAp doped NPs rendered scaffolds with
different properties, as compared to pure PCL.

Tensile stress failure tests showed that the scaffolds with doped HAp NPs
present a superior mechanical strength, with a stress at failure of 3.33MP, while
pure PCL was found to be 1.3 MPa.

The authors hypothesized that this improvement is a result of the incorporation of

HAp NPs, similar to what happens in native bone where HAp nanocrystals
increase its stiffness and compressive strength.

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Silk fibroin (SF), combined with calcium phosphate NPs, is also being used as
scaffold.
L. Yan et al. have developed a new method for silk fibroin/nanosized calcium
phosphate scaffolds. The resulting scaffold displayed higher porosity,
osteoconductivity, and osteogenic potential.

In vivo studies on rat femur showed that this scaffold induced new bone
formation after 3 weeks of implantation, compared to bare SF Scaffolds

Roohani-Esfahani et al. replaced the HAp NPs by bioactive glass NPs to

increase the degradation rate and to enhance the bioactivity of biphasic calcium
phosphate scaffolds coated with HAp–PCL composites.

This strategy also led to a significant increase of the gene expression and bone
sialoprotein in human osteoblast cells cultured for 7 days.

Hafezi et al. designed and tested in vivo a gelatin/bioactive glass NPs scaffold.

After 12 weeks of application in an defect on rabbits, radiographs of the defect

site have shown that the composite scaffold helped bone growth and healing.

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