INTRODUCTION

3D printing was originally developed in 1984 by Chuck Hull and is an additive

process of making a three-dimensional solid object from a digital model. 3D printing
is achieved by laying down successive layers of material to form shapes. There has
been recently a huge growth in the sales and use of 3D printing and the market for
these increased by 29% from 2011 to year 2012. To print, the machine reads the
design from an .stl file and lays down successive layers of material to build a series of
cross sections. These layers, as decided by the CAD model, are joined or
automatically fuse to create the final shape. This technique allows for the ability to
create almost any shape or geometric feature.The printer resolution (layer thickness
and X-Y resolution) are defined as either dpi (dots per inch) or micrometers. Typical
layer thickness is around 100 micrometers (m). Higher end machines however
(Objet Connex series and 3D Systems' ProJet series) can print layers as thin as 16 m.
The X-Y resolution of 3D printers is around 50 to 100 m.Among other uses, 3D
printing has found promise in the biomedical field as a means to generate tissue
scaffolds out of biodegradable polymers as well as potentially tissues by printing cells
and matrix into a defined area.
Current synthetic bone grafts suffer from poor handling characteristics, brittle
mechanical properties, and inconsistent bioactivity. By blending a biodegradable
amphiphilic polymer with hydroxyapatite (HA), the main mineral component in bone,
we developed an improved synthetic bone graft. The polymer/HA composites were
fabricated in both 2-D and 3-D forms by electrospinning and 3-D printing. These
materials exhibited unique handling characteristics such as high tensile elasticity
(>200% failure strain) and self-stiffening properties upon hydration, allowing their
facile/stabile fixation around an open defect or within a confined defect. They are also
superhydrophilic, enabling the absorption of aqueous cell suspensions and protein
therapeutics.

Design Properties
Low mechanical strength is a major challenge in porous scaffolds, and is
primarily controlled by pore volume and distribution. This is also true for 3D printed
ceramic scaffolds and limits their use to only non-load bearing and low-load bearing
applications. Optimized post processing approaches and compositional modifications
can improve mechanical properties of ceramic scaffolds. To investigate the effect of
printing orientation and layer thickness on the mechanical properties of green
specimens, compression tests were performed on raw Zp 150 specimens.
The compression strength, Young's modulus and toughness of the 3D printed porous
samples were determined and compared with each other to find the optimum printing
conditions. The same test setup and parameter were used for all other samples.
Calcium Sulfate scaffolds not subjected to any post hardening demonstrated lower
compressive strength, compressive modulus and compressive toughness than those
reported for cancellous bone. The compressive stress-strain curves shown in Figure
13(ac) are characterized by the initial non-linear toe region followed by the main
linear region and then the concave shape till the failure point. The fluctuations
observed in this region could be attributed to the layer by layer collapse of the
microstructure under compression load. The average values of compressive strength,
Young's modulus and toughness of five samples were shown in Figure 14(ac).

Figure 13. Compressive Stress-Strain Curve for different layer thickness in X (a), Y
(b) and Z (c) direction printing.

Figure 14. Comparison of compressive strength (a), Young's modulus (b) and
toughness (c) in samples printed with different layer thickness in various orientation.
According to the compressive stress-strain curves in Figure 13(ac), scaffolds
underwent the elastic displacement followed by failure in struts and microcracks
generation in the periphery wall of the scaffolds printed in X and Y direction through
the horizontal struts. Failure also occurred through the vertical struts in the body of
samples printed in Z direction. Moreover, the location of failures is not concentrated
in the middle of the scaffolds. This indicates that internal structure has a significant
influence on the mechanical properties of 3DP samples.
As shown in Figures 14(ac), difference in printing orientation resulted in various
compressive strengths. The compressive strength of samples printed in Z direction
was found to be very low for the printed porous samples that is critical for the
depowdering and handling steps [30]. Scaffolds printed in X and Y direction with
layer thickness of 0.1 mm had the less compressive strength, however, increasing
layer thickness from 0.0875 mm to 0.1125 and 0.125 mm had a more positive effect
on the mechanical properties of the scaffold.
As shown in Figure 14(a) and 14(c), although the samples with layer thickness of
0.0875 mm printed in Z direction have the least compressive strength but exhibit more
toughness compared to samples with 0.1 mm layer thickness.
As shown in Figures 14(a) and 14(c), scaffolds with layer thickness of 0.1 mm
demonstrated low compressive strength, Young's modulus and toughness in both X
and Y printing orientations. By increasing the layer thickness to 0.1125 and 0.125
mm, the compressive strength increased, and the plastic region was extended,
suggesting higher toughness in three orientation of X, Y and Z. Conversely, the
scaffolds printed with 0.1 mm layer thickness demonstrated a lower level of plastic
deformation and generally failed shortly after reaching the peak load that is more
evident in Z printing orientation, (Figure 13(c)). As it can be seen, the greatest
improvement in compressive strength and toughness were all obtained when scaffolds
were printed with 0.1125 mm layer thickness in X printing direction.

It seems that an increase of layer thickness and decrease of shear forces result in
better powder spreading, stability and uniformity which improve the strength.
However, when layer thickness decreases, the number of layers is increased. This may
results in higher integrity that in turn will increase the strength of the specimens too
[23]. It is also worthy to note that, as shown in Figure 14(a), under the same binder
saturation, with a decrease of layer thickness from 0.1 to 0.0875 mm, the compressive
strength would somehow increase. In such case, as layer thickness decreases, the
sprayed binder would penetrate better in vertical and lateral directions over the
surface resulting in less empty spaces between powder particles and increasing the
strength of the specimen. Generally, binder spreading in vertical direction is more
than that in lateral direction. So, the vertical direction will be saturated with the binder
before the lateral binder spreading is complete. However, it seems that when the
selected layer thickness is less than a certain threshold, the binder would completely
penetrate vertically and the powder gets saturated, while this is not the case in lateral
direction. So, incomplete spreading of the binder laterally would decrease the sample
integrity and strength. Furthermore, with a decrease of layer thickness from 0.125 and
0.1125 to 0.1 mm, the binder penetrates faster to the bottom of the layer. However, the
previous printed layer prevents the binder from further spreading which results in
nonuniformity in the interface layers. Therefore finding the optimum layer thickness
is critical for printing such porous scaffolds.
The mechanical behavior depends on the orientation of the powder spreading and
binder jetting. For scaffolds printed in Y direction the compressive load was applied
parallel to the constituent layers and the direction of binder jetting. While for those
samples printed in X orientation, the compressive load was applied parallel to the
constituent layers but perpendicular to the direction of binder jetting. For samples
printed in Z orientation, the compressive load was applied perpendicular to both, the
constituent layer and the binder jetting.
Scaffolds printed in X orientation present higher compressive strength and modulus in
comparison with the scaffolds printed in Y and Z directions. These results suggest that
the printing orientation and layer thickness have a great influence on the mechanical
properties of 3DP parts. However, these results are in contrast to the study result of M.
Castilho et al.
According to Figure 14(a), the weakest average compressive strength was shown by
the samples printed in Z direction. Also, more average strength was observed in
samples printed in X direction and this set also showed the lowest standard deviation.
The samples printed in Y direction have the mean compressive strength. Although,
this set showed the highest standard deviation referring to the significant diversity
among strength values.
It should be noted that due to the low strength of samples printed in Z direction, they
broke in the depowdering step.

TECHNIQUE TO FABRICATE BIOMATERIALS

1) 3D Plotting / Direct Ink Writing
Strands of paste/viscous material (in solution form) extrusion based on the
predesigned structure
Layer by layer deposition of strands at constant rate, under specific pressure
Disruption of strands according to the tear of speed
2)

Laser-assisted bio printing (LAB)

Coating the desired material on transparent quartz disk (ribbon)
Deposition control by laser pulse energy
Resolution control by distance between ribbon/substrate, spot size and stage
movement

3)

Selective Laser Sintering (SLS)

Preparing the powder bed
Layer by layer addition of powder
Sintering each layer according to the CAD file, using laser source

4) Fascial Distortion Model (FDM)

Strands of heated polymer/ceramics extrusion through nozzle
5) Robotic Assisted Deposition
Direct writing of liquid using a nozzle
Consolidation through liquid-to-gel transition

SPECIFIC APPLICATION OF BIOMATERIALS

Bone grafting is a surgical procedure that replaces missing bone in order to repair
bone fractures that are extremely complex, pose a significant health risk to the patient,
or fail to heal properly.
Bioceramics have been considered for use as synthetic bone graft substitutes for over
30 years, throughout which there have been two primary areas of research:
(i)
optimization of the physical pore structure and
(ii)
formulation of an appropriate bioceramic chemistry.
While it is well recognized that both the rate of integration and the final volume of
regenerated bone are primarily dependent on the macroporosity, there will still to be
some dispute regarding the optimum type of porosity. The rate and quality of bone
integration have, in turn, been related to a dependence on pore size, porosity volume
fraction, and interconnection size and interconnection density, both as a function of
structural permeability and mechano-transduction. The role of strut microstructure
and pore geometry have been considered with respect to influence on entrapment and
recruitment of growth factors in addition to its influence on scaffold mechanics.
Deconvoluting the relative affects of these parameters is complicated by the use of
both resorbable and nonresorbable bioactive bioceramics, which are believed to
mediate bioactivity in the osseous environment through two principal mechanisms:
(i)
directly through dissolution and release of ionic products in vivo, elevating
local concentrations of soluble species that interact directly with local cells
or influence cell behavior by their effect on local pH,
(ii)
(ii) indirectly through the influence that surface chemistry will have on
protein adsorption, GF entrapment, and subsequent cell attachment and
function.
One of the health applications of 3D printing most often discussed is the
manufacturing of bespoke scaffolds that could be used to mend broken bones. It is
a promising idea, but one that has been held back by the difficulties in printing
materials that are strong, flexible and can encourage the regrowth of healthy bone in
the same solution as current methods, such as bone grafts. One material being
investigated for making these 3D-printed scaffolds is calcium phosphate after all it
is a primary constituent of naturally-formed bone. The problem however is that
current 3D printing methods to make calcium phosphate scaffolds require the use of
high temperatures. Such temperatures make it impossible important drugs which
can stimulate bone growth, antibiotics to prevent infections, or even living cells to
the scaffolds. At the moment, calcium phosphate powder is temporarily bound using
an acidic binder chemical typically phosphoric acid and then permanently fused
together by sintering. Aside from the temperature issue, scaffolds made in this way
sometimes retain unwanted binder residues, while during sintering, parts shrink
sometimes in an uneven way that can cause cracking. Now, researchers in the U.S.
have developed a new way of printing in calcium phosphate that can be done at
relatively low temperatures which results in a scaffold that is mechanically strong,
biologically compatible with human bone and stays close to the original dimensions
of the design being used.Their approach relies on adding collagen to the phosphoric
acid binder solution to create a composite material with the calcium phosphate, they

report in the journal Biomaterials. Collagen is the most abundant structural protein
in the body and is a critical component of bone, adding to its strength and
toughness. The team from the University of Rochester, Alfred University and the
University of California San Diego also added a surfactant to the mix to improve
the printability of the material. They printed scaffolds using the composite material
on an adapted 3D Systems ZPrinter 450 and subjected them to a series of
laboratory tests. They found the collagen composite scaffolds were tougher than
those made using calcium phosphate alone, and were also better at encouraging the
growth of living cells. Moreover, the improved scaffolds were as effective in
bridging bone defects in animal models as regular bone grafts, but in common with
prior studies of 3D printed scaffolds, were unable to stimulate complete healing.
Bone graft composite can be designed and fabricated using additive manufacturing
(AM) approaches. Different AM approaches other than 3D printing (3DP) are Solid
Freeform Fabrication (SFF) also known as Rapid Prototyping (RP). These approaches
allow complex shapes for scaffolds fabrication directly from a Computer Aided
Design (CAD) file.
3D printing refers to any of the various processes for printing a threedimensional object. Primarily additive processes are used, in which successive layers
of material are laid down under computer control. These objects can be of almost any
shape or geometry, and are produced from a 3D model or other electronic data source.
While Rapid Prototyping is a group of techniques used to quickly fabricate a scale
model of a physical part or assembly using three-dimensional Computer Aided Design
(CAD) data. Construction of the part or assembly is usually done using 3D printing or
"additive layer manufacturing" technology.
3D printing starts with making a virtual design of the object you want to
create. This virtual design is made in a Computer Aided Design (CAD) file using a 3D
modeling program (for the creation of a totally new object) or with the use of a 3D
scanner (to copy an existing object). This scanner makes a 3D digital copy of an
object and puts it into a 3D modeling program. To prepare the digital file created in a
3D modeling program for printing, the software slices the final model into hundreds
or thousands of horizontal layers. When this prepared file is uploaded in the 3D
printer, the printer creates the object layer by layer. The 3D printer reads every slice
(or 2D image) and proceeds to create the object blending each layer together with no
sign of the layering visible, resulting in one three dimensional object. Meanwhile,
Rapid Prototyping process begins with taking a virtual design from modeling or
Computer Aided Design (CAD) software. The 3D printing machine reads the data
from the CAD drawing and lays down successive layers of liquid, powder, or sheet
material building up the physical model from a series of cross sections. These
layers, which correspond to the virtual cross section from the CAD model, are
automatically joined together to create the final shape. Rapid Prototyping uses a
standard data interface, implemented as the STL file format, to translate from the
CAD software to the 3D prototyping machine. The STL (STereoLithography) file
approximates the shape of a part or assembly using triangular facets.
One of the main advantages of 3D printing is that it allows the manufacturing
of objects having complex geometries and intricate internal structure, which can be

designed according to the needs of individual patients using their 3D medical scan
data. A great prospect for biomedical applications, especially for tissue engineering
applications, has been shown. Both natural and synthetic polymers have been
developed for tissue engineering via the 3D printing technology, and numerous
additional materials are being developed. Rapid Prototyping increase visualization
capability during the early phases of design by using rapid physical model, detect
design flaws before the manufacture of tooling, rapidly create tooling to manufacture
physical prototypes and cost savings were achieved on the development project.
But this form of 3D printing has two major limitations today. One is that the
resulting objects have little tensile strength and little temperature tolerance. They're
fine for figurines and other objects d'art, but they're not able to handle the stress of
holding up a load or of other pressures, nor do they maintain their shape in heat or
stay intact in extreme cold. Some sources are of the opinion that rapid prototyping is
not effective because, in actual, it fails in replication of the real product or system. It
could so happen that some important developmental steps could be omitted to get a
quick and cheap working model. This can be one of the greatest disadvantages of
Rapid Prototyping. Another disadvantage of Rapid Prototyping is one in which many
problems are overlooked resulting in endless rectifications and revisions. One more
disadvantage of rapid prototyping is that it may not be suitable for large sized
applications.

Conclusion
In conclusion, there are several ways to produce an artificial bone graft. In this
assignment, we focus on the technique of 3D printing. The use of low temperature in
3D printing by using the materials from calcium phosphate and collagen be an
alternative for artificial bone graft fabrication. The technique of 3D discussed in this
assignment is similar to what we are familiar with the makerbot in the market to
create the object. Their process differs with the type of printer and material used.
Calcium phosphate is chosen to be the core material due because its biocompatibility
and similarities to real bone. Next, the collagen is used to promote cell growth of the
human cells cultured on the surface of the 3D printed object. By adding collagen to
the calcium phosphate and binding material before printing, cell viability and the
strength of the final object can be increased.
3D printing offers unique advantages toward part fabrication that are need for
the production of small volumes or one of a kind product manufacturing. Compare to
other manufacturing techniques, 3D printing is a versatile tool that has become
popular for making scaffolds in bone tissue engineering. 3D printing can fabricate
scaffolds with defined shapes, with controlled and interconnected porous structures.
Although it might fabricate almost all types of materials, the selection of suitable
binder for 3D printing is still a challenge. The residue from binders may be difficult to
remove during sintering, an issue that may need special attention for biomaterials.
In terms of future work and marketable view, it is expected that the demand
for processes such as 3D printing will be increasing in the coming years due to their
ability to make custom medical devices that can be mould or shape for specific patient
needs and clinical defects. Experts have forecast that the market value of 3D printing
will be increasing in the future. However, extensive process-property optimization is
still needed to accomplish this goal. For ceramics, the most critical issue that needs
attention is the mechanical properties of porous scaffolds. Resorbable polymer
infiltration to enhance strength and toughness in these scaffolds is one way to
minimize this problem; the use of resorbable glassy materials can also help. Although
current techniques let us build structures with similar composition to that of tissue, we
are still a long way from completely printing functioning tissue. More processproperty optimization, in vitro and in vivo research are needed in that direction to
make any of those approaches useful toward bone tissue engineering.

References
Jason A. Inzana, Diana Olvera, Seth M. Fuller, James P. Kelly.(2014).
3D printing of composite calcium phosphate and collagen scaffolds for
bone regeneration. Biomaterials 25 (2014) 4026-4034
Susmita Bose, Sahar Vahabzadeh and Amit Bandyopadhyay.(Dec
2013).Bone tissue engineering using 3D printing.Materials Today.
16(12).
Natalja E. Fedorovich, Jacqueline Alblas.(Dec 2011). Organ printing:
the future of bone regeneration?.Trends in Biotechnology. 29(12).
Sopyan, M.Mel, S. Ramesh.(2009). Porous hydroxyapatite for artificial
bone applications. Science and Technology of Advanced Materials.
8(2007). Pp. 116-123
Molly M. Stevens.(May 2009). Biomaterials for bone tissue
engineering. Materialstoday. 11(5)

APPENDIX