Multilayered scaffold for temporally

controlled release of growth factors in the

treatment of Periodontitis

Madhumathi.K
BT5130
Dept of Biotechnology
IIT Madras

Periodontitis
An infectious and inflammatory
disease resulting in destruction of
periodontium and loss of teeth.
Common bacterial infection causing
tooth destruction

Periodontium
Periodontium consists of
gingiva (gums),
alveolar bone (part of jaw bone covering the tooth
socket)
cementum (a non vascular connective tissue covering
the root of the teeth) and
periodontal ligament fibers (connective tissue fibers
that connect alveolar bone to cementum).

Periodontium is referred to as the attachment

apparatus since it serves as an anchor for the
teeth.

Periodontium

Periodontium
Types of cells:
Periodontal ligament cells (PDLs)
fibroblasts
cementoblasts
osteoblasts and
Periodontal ligament stem cells (PDLSC).
Signal molecules/growth factors:
Bone morphogenic protein (BMP)
fibroblast growth factor (FGF)
platelet derived growth factor (PDGF)
insulin-like growth factor-1 (IGF-1) etc.

Current treatment
a) cleaning the root surface to remove the
plaque
b) priming the exposed root surface with
fibronectin to facilitate attachment of cells
b) administration of BMP for bone regrowth.
c) placement of barrier membranes made of
Cellulose, collagen, poly lactic acid and poly
glycolic acid

Failures in current approach

Unpredictable
Only partial regeneration is observed
Low
regenerative
potential
of
periodontium
Complex process involving epithelial,
bone and connective tissue engineering

Periodontal tissue regeneration

Cells

Scaffold

Growth
factors

Periodontal tissue regeneration

Current tissue engineering approaches:
Focuses more on scaffold and cellular aspect.
Growth factors delivered as topical delivery.
Delivery of single growth factor.

INCOMPLETE PERIODONTAL REGENERATION

Sequential Growth factors Delivery

Growth factors have short half-life
Should be delivered at low concentrations so
as to not disrupt natural process.
Multiple growth factors necessary to achieve
complete regeneration.
These growth factors can negatively regulate
at high concentrations or when administered
over long period

Scaffold design
The periodontium has bone on outer side
and soft tissues on inner side.
Multilayered Scaffold design
Side facing bone to be osteoconductive
Part of scaffold facing the soft tissues
should be designed to mimic fibrous
component embedded in extracellular
matrix (ECM).
Designed to release multiple growth
factors

Work Plan
Fabrication of multilayered fibrin-collagen/BCP
scaffold
Characterization of fabricated scaffold
In-vitro studies:
Biocompatibility
Osteoconductivity
Growth factor release assay

In vivo studies
Clinical trial

Biomaterials for scaffolds

BCP Biphasic calcium phosphate[ Hydroxyapatite + Tricalcium
phosphate] to form the outer layer.
This bioceramic layer to act as depot for growth factors- growth
factors release based on degradation.
Fibrin matrix with collagen fibres to mimic natural PDL during
wound healing.
Fibrin is an end product of coagulation pathway - rich in growth
factors and platelets.
Fibrin can be prepared from patients own blood thereby
eliminating problems of immunogenicity, allergic responses etc.

Fabrication of scaffolds
a) Fabrication of BCP construct.
Synthesis of BCP powder
Formation of growth factor loaded BCP construct

b) Fabrication of Collagen scaffold.

Electrospinning

c) Fabrication of fibrin rich bilayered construct.

Fabrication of BCP Construct

Calcium hydroxide + Diammonium hydrogen phosphate
Ca/P ratio = 1.5

CDHA

Ca/P ratio =1.67

TCP

HA

heated to 700C

Collagen+ Growth factors

BCP Powder

BCP
Collagen+ Growth factors
(BCP and TGF-)

BCP Construct

Fabrication of Collagen Layer

Collagen dissolved at various concentrations in
1,1,1,3,3,3 hexafluoro-2-propanol (HFP)
Electrospinning

Electrospun Collagen

BCP construct with

growth factors
Electrospinning set up

Fabrication of fibrin rich bilayered construct

The fibrinogen solution (6 mg/ml, 0.8 ml) containing growth factors like
PDGF and FGF is poured into a well containing scaffold.
Then, 4 IU of thrombin solution (0.8 ml) is poured into each well to set
a fibrin gel.
This followed by adding 50ml CaCl2 resulting in polymerized fibrin gel
after a period of 20min at RT.

Fibrin
Electrospun Collagen

BCP construct with growth factors

Alternate method
Fabrication of growth factor loaded
collagen microspheres in BCP matrix

Method of fabrication of microspheres : oil in

water emulsion

Characterization
FT-IR
XRD

SEM

TEM
Porosity

Scaffold

Peeling
test

In-vitro studies
Biocompatibility : Alamar blue Assay
Osteoconductivity: Biomineralisation study in
SBF
Cell adhesion studies: culturing of hPDLF cells
Growth factor release assay by ELISA

Alternate solution
Fabrication of periodontal ligament stem cell loaded
fibrin-collagen/BCP scaffold.
Increases local cell population by differentiation.
Extraction of
stem cells
from teeth
Culturing in
Eagles
medium
Seeding of
stem cells
onto scaffold

In-vivo studies
Animal studies:
Dogs/Pigs can be used for evaluating the
biocompatibility
and
regenerative
potential of periodontium
At the end of 3 months, the animals have
to be sacrified and tissue sections have to
be studied to observe the extent of
periodontal regeneration.

Clinical trial
Clinical trials have to be performed on human
volunteers.
The group can be divided into three categories,
control group (no scaffold), scaffold without growth
factors and scaffold with growth factors.
The trial has to be conducted for a period of 3-6
months to observe the regeneration. The
regeneration can be evaluated both clinically and
radiographically.

Timeline
Period of study

Target

6 Months

Fabrication of scaffold

18 months

30 months

Optimization for controlled

release of growth factors
Characterization and in
vitro studies
In vivo animal studies

36 months

Clinical trial

24 months

Questions??