BIOPRINTING

Regenerative Medicine

SAMEEN AHMED MALIK

Department of Biomedical Engineering
Faculty of Engineering
University of Engineering & Technology Lahore
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Definition of Bioprinting

• Bioprinting is a technology where bioinks and

biomaterials, mixed with cells, are 3D printed to
construct natural tissue-like three-dimensional
structures.
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History of Bioprinting
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Why Bioprint Organs

• The demand for organ transplantation has rapidly grown over the
past two decades.
• Worldwide increases in organ failure combined with rising successes
in post-transplantation outcomes has resulted in an organ shortage
crisis, as the demand for organs vastly outstrips supply.
• Post-operatively, the immune system of the transplant recipient may
recognize the new organ as ‘foreign’ and trigger transplant rejection.
• 3D bioprinting is used for developing in vitro models for testing
pharmaceutical drugs
• Potential to minimize lengthy and expensive clinical trials and save
billions of dollars.
• Bioprinting can save lives by providing patients with personalized
organs made from their cells, lowering the risk of organ rejection.
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How Bioprinting Works

• 3D bioprinting is an additive manufacturing

process where organic and biological materials
such as living cells and nutrients are combined to
create artificial structures that imitate natural
human tissues.
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How Bioprinting Works

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How Bioprinting
Works
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Steps in 3D Bioprinting

• Pre-printing/pre-processing. This includes

selection of cells and bioink materials, mixing of
the ingredients, and creating the 3D design.
• Bioprinting using one or more of the available
printing technologies to bioprint a tissue
construct and crosslinking to maintain a stable
structure.
• Post-printing. This phase includes cultivation of
the tissue structure in a bioreactor.
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Steps in 3D Bioprinting
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Steps in 3D Bioprinting
Pre-processing (Designing)

• The first step is to conceptualize and

specify the desired structure.
• This will define the shape, size, and
general physical properties of the tissue
and the types of cells and
support/nutrient materials that will be
used.
• With the outline of the structure defined,
specialist CAD tools Solidworks etc are
used to create a highly detailed 3D model
of the structure.
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Steps in 3D Bioprinting
Pre-processing (Cell Preparation)

• An appropriate bio-ink is then selected or created,

containing the structural, hardening, and nutrient
mix appropriate for the tissues to be grown.
• Cell selection and cultivation in vitro is the most
delicate step, involving incubating and
encouraging the reproduction of cells in a culture
medium under tightly controlled conditions, to
ensure their viability and sufficiency.
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Steps in 3D Bioprinting
Pre-processing (Bioink)

• Bio-ink formulations are typically made of a combination

of biomaterials that help cells grow and mature as tissue.
• These biomaterials usually include different biopolymers,
proteins and growth factors.
• Biopolymer gels that act as a 3D molecular scaffold so
that cells can attach, grow, and increase.
• Biopolymer are essential as they retain water which
provides mechanical stability to the engineered tissues.
• The selection of bioink for a particular process is an
important step as the selected bioinks should have
desired physicochemical properties that include
mechanical, chemical, biological, and rheological
characteristics.
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Steps in 3D Bioprinting
Pre-processing (Bioink)
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Steps in 3D Bioprinting
Processing

• The second step is the actual printing process

where the bioink is placed in the printer to form a
3D structure.
• The mixture of cells, nutrients, and matrix,
together forming bioink, is then placed onto the
printer cartridge, which then deposits the
material based on the digital model prepared.
• The formation of biological constructs involves
the deposition of bioink onto the scaffold in a
layer-by-layer approach to generate a 3D tissue
structure.
• This step of the bioprinting process is a complex
process as it requires the formation of different
cell types based on the type of tissues and organs
to be formed.
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Steps in 3D Bioprinting
Post-processing

• This step is important to provide stability to the printed

structure.
• Crosslinking can use different environmental controls like
UV light, temperature, and chemicals, among others to
maintain structure and function of the biological matter
• Finally, the bioprinted and crosslinked structures need to
grow biologically.
• This means that the printed living cells will reproduce, and
tissue will grow following the underlying printed
structures.
• This step is also called incubation and is done inside
bioreactors that create a favorable environment for
reproduction and tissue growth.
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Steps in 3D Bioprinting
Evaluation

• Structural Quality
• Mechanical Stability and Elasticity
• Swelling and Degradation
• Effect of the Printing Process on Cell Viability
• Cell-Matrix Interactions within 3D Printed
Structures
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Steps in 3D Bioprinting
Evaluation
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Steps in 3D Bioprinting
Evaluation
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Bioprinters

• 3D printers and 3D bioprinters are similar to each other,

but 3D printers are designed to print solid materials,
where 3D bioprinters are designed to print liquid or gel.
• 3D bioprinters are also designed to handle sensitive
material that contain living cells, without creating too
much damage on the end result.
• Bioprinters can be inkjet based, laser assisted, or
extrusion based. Each printer type has its pros and cons
when it comes to cost, cell viability, cell density,
resolution, and so on.
• Bioprinters’ compatibility with bioinks also varies, and
therefore it is important to ensure the bioprinter and
bioink work well together.
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Bioprinters
Types

• Inkjet Bioprinting
• Extrusion-Based Bioprinting
• Laser Assisted Bioprinting (LAB)
• Stereolithographic Based 3D Bioprinting
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Bioprinters
Types (Inkjet Bioprinting)
• Inkjet bioprinting is based on the ejection of drops of liquid
onto a substrate by thermal or acoustic forces.
• Thermal inkjet bioprinting can be achieved by electrically
heating the print head to generate pressure that causes the
release of droplets from the nozzle.
• In the case of acoustic inkjet bioprinting, a piezoelectric crystal
is used that creates an acoustic wave inside the print head to
break the liquid into droplets.
• When a voltage is applied to a piezoelectric substance, a rapid
change in shape is induced. This, in turn, generates pressure
required to force droplets out of the nozzle.
• Both of these methods have their own advantages and
disadvantages; thus, the selection of inkjet bioprinting
technology should be made based on the desired purpose.
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Bioprinters
Types (Inkjet Bioprinting)
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Bioprinters
Types (Extrusion Bioprinting)

• Extrusion-based bioprinting utilizes a computer-

controlled system to continuously extrude bio-
inks, specifically viscoelastic biomaterials in
filaments.
• Using layer-by-layer extrusion with the nozzle
free to move in the x-y-z directions and an
adjustable printer stage to fabricate a 3D
construct.
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Bioprinters
Types (Extrusion Bioprinting)
• The pneumatic-based technique uses pressured air at a controlled
volume flow rate to drive fluid dispensing systems’ constant
extrusion of bio-ink.
• The piston/screw-based technique mechanically forces biomaterials
out of the nozzle.
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Bioprinters
Types (Laser Assisted Bioprinting)
• Laser-assisted bioprinters were first introduced in 1999 by
David Odde using optical cell trapping.
• Laser assisted bio-printing (LIB) consists of a pulse energy
laser, ribbon substrate coated with bioink and receiving plate.
• When laser pulse energy is transferred to the ribbon layer, the
energy absorbing layer is then activated, formation of high-
pressure bubbles, propelling the cell-containing material
towards the receiving substrate .
• During the printing process, the risk of contamination is low
because the dispenser and the bio-inks are not in contact.
• The main advantage of this system is that it can deposit bio-inks
with relatively high viscosity and resolution. Moreover, the
issue of nozzle clogging is eliminated because this system
involves a nozzle-free printing process
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Bioprinters
Types (Laser Assisted Bioprinting)
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Bioprinters
Types (Stereolithography Bioprinting)
• At the start, the build platform is positioned or dipped in the tank at a
distance equal to the height of one layer from the surface of the
liquid resin.
• The UV laser is then activated and falls on the scanning mirror.
• This mirror directs the laser beam on the resin and it traces the X-Y
geometry of the design or the cross-sectional area of the model.
• The resin is cured and it hardens and the first layer sticks to the build
platform. This completes the first layer of the print.
• Once one layer is printed, the platform is raised and a wiping system
sweeps through the bottom of the tank to eliminate bubbles, if any
and reduce the chances of a print failure.
• Next, the build platform drops down by one layer height and the laser
again activates and traces the geometry of the next layer. This
process continues till the complete model is 3D printed.
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Bioprinters
Types (Stereolithography Bioprinting)
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Bioprinters
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Bioprinters
Commercially Available
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Applications of 3D Bioprinting

• Drug development done ethically, fast and cost-

effective.
• Bioprinted artificial organs
• Wound healing through bioprinted constructs
• Skin tissue
• Cardiac tissue
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Benefits of 3D Bioprinting
• Allows mimicking the real structure of desired
tissue/organ etc.
• Possibility to revolutionize future medical treatment
capabilities
• Possible creation of patient-specific and organ-
specific treatments
• Effects of drugs can be examined more accurately
• Decreases animal testing
• Biocompatibility with human cells and tissues
• Automating complex processes
• Consistency, less human errors
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Limitations of 3D Bioprinting

• Material incompatibility
• Maintaining cell environment can be difficult
• Cell damage during production
• Expensive
• Ethical concerns