Tissue Engineering CHE 418/518

Graphitic Carbon Materials for

Bone Tissue Engineering

http://www.mech.kuleuven.be/en/bme/research/mechbio/

http://en.wikipedia.org/wiki/Graphene

Tissue Engineering CHE 418/518

Bone Tissue Engineering

There are approximately 500, 000 surgical procedures performed every year
in the U.S. which require bone substitutes.

http://www.cs.cmu.edu/~tissue/
1. Interconnected micro-pores (for vascular formation and waste transportation)
2. Optimal porosity with adequate surface area and mechanical strength
3. Biodegradable
4. Biocompatible

Tissue Engineering CHE 418/518

Carbon Nanotubes
Allotropes of carbon with a cylindrical nanostructure
Single walled carbon nanotubes
Multi-walled carbon nanotubes

Mechanical strength

Electric properties
metallic nanotubes
current density 4 109 A/cm2
1000 times better than copper
Biocompability ???

http://nanopedia.case.edu/

http://what-whenhow.com/nanoscience-andnanotechnology/carbon-nanotubes-andother-carbon-materials-part-1nanotechnology/

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

(1) How do the carbon nanotubes affect the pore structure/porosity and mechanical
properties of scaffolds?
(2) What are the responses of cells cultured on scaffold made of different
nanomaterials with varying porosity?

What have been done?

Bone Cell Proliferation on Carbon
Nanotubes
Nano Lett., 2006, 6 (3), pp 562567

CNTs sustain osteoblast growth and bone formation;

Surface charge/modification play an important role for
osteoblast proliferation.

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

Highly Ordered MWNT-Based
Matrixes: Topography at the
Nanoscale Conceived for Tissue
Engineering
Langmuir, 2006, 22 (12), pp 54275434

This method aims to create architectures and topographies that mimic those
occurring naturally (native tissue structures).
Colloids and Surfaces B: Biointerfaces
Volume 83, Issue 2, 1 April 2011, Pages 367-375

(a) PLGA, (b) PLGA/MWCNTs, and (c) PLGA/cMWCNTs

Possessed higher tensile strength

Promoted the attachment, proliferation, and ALP secretion of rat MSCs

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

Journal of Biomedical Materials Research Volume 59, Issue 3

Electrical stimulation (10 mA at 10 Hz) for 6 h/day for various periods of time,
46% increase in cell proliferation after 2 days,
307% increase in the concentration of extracellular calcium after 21 consecutive days,

Multiwalled carbon nanotube-coating of 3D collagen

scaffolds for bone tissue engineering
Rat primary osteoblasts (ROBs) were cultured,
Significantly more bone formation was observed
around the MWCNT-coated sponges
Carbon Volume 49, Issue 10, August 2011, Pages
3284-3291

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

What next?
3-D scaffolds comprised of CNTs with controlled
size/distribution/surface modification and their
evaluation for bone tissue engineering

Formation of 3-D scaffold

Control size/distribution/surface modification for CNTs
Incorporation in polymer
Evaluate the charge effect

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

Formation of 3-D scaffold
Conventional: Carbon nanotubes were separately synthesized and then dispersed,
Can we arrange carbon nanotubes in controlled 3-D fashion?

Nature Materials 10, 424428 (2011)

Three-dimensional flexible and conductive

interconnected graphene networks
Carbon nanotubes

Adv. Mater. 44/2009

Graphene

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

ACS Nano, 2011, 5 (8), pp 64036409

http://www.azonano.com/article.aspx?ArticleID=1405

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

The diameter is only dependent on the size of nanoparticles,
The distribution is only dependent on the density of nanoparticles,
Selectively decorating graphene layer with catalytic nanoparticles
Covalent binding between graphene and NPs

Formation of 3-D scaffold comprised of arranged carbon nanotubes

Why important? Mechanical signal
Dimension and distribution of carbon nanotubes in 3-D fashion directly decide the topography
of the scaffold, which has a great impact on proliferation and differentiation of cells.

Carbon Nanotubes for Bone Tissue Engineering

Modification of 3-D scaffold comprised of arranged carbon nanotubes
1. Hydrophobic/Hydrophilic
2. Neutral/Negative/Positive charge

-COOH
-COOH

acid

-COOH
-COOH
-COOH

Characterization
Confirmation of desired structure
1. SEM and TEM
2. FTIR for surface functional group functionalization
3. Cell culturing
Confirmation of desired surface groups
Effect of mechnical signal for growth of cells
(Cell number, differentiation, adhesion)

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

Incorporation in polymer
PLGA poly(lactic-co-glycolic acid
FDA-approved, easily processed, relatively inexpensive, biodegradable polymer
Degradation of PLGA resulted in lactic acid and glycolic acid, can be removed from the
body by metabolic pathways,
The application of PLGA has been limited by the weak mechanical strength.

Combined with 3-D scaffold comprised of CNTs and

graphene network

Surface modification + polymerization

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

Mechanical strength evaluation

Strength of composite should

be enhanced greatly.

Colloids and Surfaces B: Biointerfaces

Volume 83, Issue 2, 1 April 2011, Pages 367-375

Effect of carbon nanotubes for degradation

Degradation of PLGA can be controlled by alternating hydrophilic and
hydrophobic region, could presence of CNTs (hydrophobic/hydrophilic)
accelerate/decelerate the degradation of scaffold?
Effect of addition of 3-D scaffold for bone cell fomation
(Cell number, differentiation, adhesion)

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

Electrical stimulation

supply

Only junction of CNTs could provide

electrical signal path, localized and nonuniform

Uniform path, conclusive about

the effect of electrical signal

Tissue Engineering CHE 418/518

Carbon Nanotubes for Bone Tissue Engineering

Possible outcome
1. Formation of 3-D scaffold comprised of graphene and carbon nanotubes,
2. Evaluation of cell behavior/response with respect to this 3-D scaffold by
changing topography/surface functional groups,

3. Application of 3-D scaffold into polymer scaffold for enhancement of

mechanical strength,
4. Effect of 3-D scaffold for degradation of PLGA due to extra reaction sites,

5. Effect of electrical signal for bone cell growth,

6. Evaluation of long term effect of graphitic carbon materials for bone tissue
engineering.