3D BIOPRINTING: PRINCIPLE, TECHNIQUES AND IT’S APPLICATION IN HUMAN THERAPY

3D BIOPRINTING: PRINCIPLE,
TECHNIQUES AND IT’S APPLICATION IN
HUMAN THERAPY
Presented By: Akshita Dholakiya
B.Pharm
SMT R.D. Gardi B.Pharmacy College Nyara, Rajkot

Content
Dr. R.D Gardi B.Pharmacy college nyara, Rajkot 2
Application of 3D Bioprinting
Types of Bioink Used In 3D Bioprinting
3D Bioprinting Techniques
Basic Principal of 3D Bioprinting
History of 3D Bioprinting
Introduction
Challenges And Future Perspective

• 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.
• In other words, bioprinting is a type of 3D printing that can potentially
produce anything from bone tissue and blood vessels to living tissues for
various medical applications, including tissue engineering and drug
testing and development.
• THE FIRST INTERNATIONAL WORKSHOP on Bioprinting and
Biopatterning was held at the University of Manchester (United
Kingdom) in September 2004 and was organized by Dr. Douglas B.
Chrisey , Dr. Richard K. Everett and Dr.Nuno Reis.
1
Dr. R.D Gardi B.Pharmacy college nyara, Rajkot
Introduction

4
• In the early 1980s, Charles Hull created 3D printing, often
known as "stereolithography."Hull, who has a bachelor's
degree in engineering physics, was employed by the
California company Ultra Violet Products to create plastic
items out of photopolymers.
• Stereolithography interprets the information in a CAD file
using the STL file format, enabling these instructions to be
sent electronically to the 3D printer. The file's instructions
could additionally specify the colour, texture, shape and
thickness of the printed object.
History of 3D Bioprinting

2.Autonomous
self
assembly,
1.Biomimicry
or
biomimetics
3.Mini-tissue
building
blocks
• The base of 3D bioprinting is the layer-by-layer exact positioning of biological constituents,
biochemicals, and living cells.
• The process of 3D bioprinting is based on three distinct approaches:
Basic Principal of 3D Bioprinting

6
3D Bioprinting Techniques
• Highly functional 3D structures have been constructed using a range of 3D bioprinting processes, including extrusion-
based, light-assisted, and inkjet-based printing systems. These systems frequently use computer-aided design and
computer-aided manufacturing to create 3D structures with great accuracy.
1. Extrusion-based bioprinting
The most popular 3D bioprinting technique is extrusion. Extrusion-based
bioprinting uses pressures that are driven by air, screws, and pistons to
dispense the cells that contain the bioink through a nozzle.
1. .

2. Light-assisted Bioprinting
• Light-assisted printing also called as laser-assisted bioprinting is a bioprinting technique
that uses light to solidify a photocurable bioink.

3. Inkjet-based Bioprinting
 The first bioprinting study, known as inkjet-based
printing, disperses cells and biomaterials to create
2D live cell patterns using a modified office inkjet
printer. This bioprinting technique can produce
and precisely apply picoliter amounts of bioink
(1–100 pL) to a substrate.

7
1. Cell-Laden Hydrogel Bioink
As they are simple to create for extrusion-based, droplet-based
(inkjet), and laser-based bioprinting methods, cell-laden
hydrogels are the most widely used bioinks. Formulations for
cell-laden hydrogel bioinks use a combination of natural and
synthetic hydrogels, including agarose, alginate, chitosan,
collagen, gelatin, fibrin, and hyaluronic acid, PGA.
Types of Bioinks Used In Bioprinting
• The ideal bioink formulation should satisfy certain material and biological requirements.
Material properties are printability, mechanics.

• Synthetic hydrogels often have good mechanical qualities in comparison to natural
hydrogels. In order to generate cross-linking (chemical- and/or photo-cross-linking)
or extra bioactivity
8

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2. Cell Suspension Bioink
• Modified inkjet printers have long been used to print cells into cellular assemblies. For
instance, endothelial cells were printed from cell suspension in growth media.
• The bioink formulation undergoes a fully biological self-assembly without or in the
presence of a temporary support layer. This technique relies on tissue liquidity and
fusion, which allow cells to self-assemble and fuse due to cell–cell interactions

3. dECM-Based Bioink
• Decellularized extracellular matrix-based bioinks include eliminating the cells from a
tissue of interest while leaving the ECM in place. To create the bioink, the ECM is then
ground into a powder and soaked in a buffer solution that is favourable to cells.
10

• Today, 3D bioprinting is successfully used in a variety of tissue engineering and regenerative
medicine applications, including the creation of disease models from hard tissue and soft tissue.
1. Biofabricated in vitro models for studying infectious diseases
 Organoids
Organoids, made up of many cell types, have been designed to morphologically and
functionally reproduce human organs. Human organoids have been effectively used in recent
years to identify the pathophysiology following either bacterial or viral infection due to this
advantage Human proximal airway organoids have shown their capacity to distinguish between
the poorly human-infecting viruses, such as the avian- infecting influenza virus and the swine-
infecting influenza virus, and the human infecting influenza virus.
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Application of 3D Bioprinting

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 Microfluidic organs-on-chip
• Numerous organ types have been studied for infectious disorders using microfluidic
organs-on-chips.
• Hepatitis B virus infection was studied using a human liver-on-a-chip to replicate
hepatocyte organization on liver sinusoids using a microfluidic recirculation system.

2. 3D Bioprinted cell and drug delivery system
• Due to the limited solubility of the majority of chemotherapy medicines, such as 5-fluorouracil, paclitaxel,
and cisplatin, the necessary drug concentration cannot be delivered to the infected site using traditional
medication approaches (such as intravenous injection and oral medication). Even worse, the systemic toxicity
and post-infusion cytokine release syndrome of such medications harm the entire organism.
 Cell delivery system
In addition to the development of cell encapsulated scaffolds in tissue engineering studies, there are some
macrophage- and antibiotic-laden delivery systems that reduce infection after surgery. Aldrich et al.
presented a 3D printed composite scaffold with antibacterial efficacy for treating bone infections after
craniotomy
15

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 Sustained drug release system
Systems loaded with antibiotic medications are also
necessary following internal organ surgeries. Various
geometries of the administration site were used to produce
the polymeric patch.

 Aerosolized delivery system
• Non-invasive mucosal administration of biologics has been developed along with local
direct delivery at the surgical site. A 3D-printed MucoJet device was created by Aranet et al.
for oral vaccination.
17

3. Bioprinting in engineering vaccines and
therapeutics
• Vaccines are biological agents that stimulate the immune system
to defend against contagious germs and viruses. The best medical
strategy for preventing and managing infectious diseases at the
moment is vaccination. There are typically three types of
vaccines: nucleic acid vaccines, subunit/conjugate vaccines, and
whole-pathogen vaccines.
 RNA printer (CureVac)
• A proof-of-concept for the mRNA printing facility has been
created for pandemic responses, allowing for the quick
formulation and delivery of vaccines. The idea is that the RNA
printer would create a vaccine that is ready for use in a clinic after
the creation and validation of several RNA vaccines CureVac, a
pioneer in mRNA printing technologies, creates mRNA vaccines
for a variety of viral illnesses, such as yellow fever, Lassa fever,
MERS, and COVID-19 using traditional production techniques.
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 Drug printer
• The pharmaceutical industry adopted 3D printing in the hopes of
creating patient-centered dosages based on structural designs. The
controlled release characteristics of the 3D printed tablets allowed
patients to take less amounts more frequently as part of their
regular routine.
• Aprecia Pharmaceuticals reformulated the anti-epileptic medication
levetiracetam, named Spritam.The first 3D printed tablets that
disintegrate within seconds in an aqueous solution were developed
through a proprietary powder bed and inkjet 3D printing
technology known as Zip Dose.

4. In tissue engineering and regenerative medicine
• Patients with valvular heart disease frequently need to have their valves replaced, either
mechanically or biologically. However, problems like mechanical failure and calcification
are usually connected to these prosthetic valves. As a result, a variety of strategies,
including the application of bioprinting technology, have been suggested to enhance the
results.
Blood vessel construct fabricated by extrusion-based bioprinting

• The skin is another area where bioprinting technology has been found to use. In one work,
a jetting-based bioprinter was used to produce biomimetic multilayered skin tissue made
of keratinocytes and fibroblasts.
• In a similar, we have created an in situ skin printer that can print skin cells directly onto
the body to treat severe burn wounds. We were able to repair pigs' full-thickness wounds
using this skin printer by supplying keratinocytes and fibroblasts, and the wounds quickly
reepithelialized and healed more quickly.
Confocal microscopic image of multilayered skin structure fabricated by jetting-.based bioprinting

• The most difficult aspects of 3D bioprinting are finding the right
materials and providing the cells with enough nutrients, which has
slowed down the development of this technology for years.
 Material selection
• Based on the method of manufacturing, the various biomaterials
used in 3D bioprinting can be broadly divided into two categories
natural polymers like collagen, chitosan, and hyaluronic acid, and
synthetic polymers like polylactic acid (PLA), polyglycolic acid
(PGA), and poly(lactic-co-glycolic acid) (PLGA).
Challenges

• Natural biopolymers typically perform poorly in terms of mechanical properties despite having
strong biocompatibility, which allows them to attain the needed formability as a single printed
material Synthetic biopolymers, in contrast, typically have poor biocompatibility and good
formability.
 Nutrient supply
• The inability to create an effective microvasculature is now a challenging issue in the creation
of in vitro large-size organs, making it challenging to provide nutrition to the cells in the
interior region of the constructed organ.
• One modern technique involves creating some channels inside the bio-fabricated organs,
provided adequate nutrients across them to simulate vasculature, and then injecting endothelial
cells, smooth muscle cells, or fibroblast cells to create vascular structures However, there is still
a ways to go before we have fully vascularized structures.

• There have been great advances on bioprinting technologies. Future
work would be the development of facile and portable bioprinting
devices that can be used in clinical settings. For example, a handheld
printer was recently developed for the fabrication of skin tissues.
• The device was easy to operate and capable of printing multimaterial
bioinks. Bioprinting technologies can also be adapted to do in
situ bioprinting. A recent example showed a laser-assisted bioprinting
for in situ bioprinting of mesenchymal stromal cells in collagen and
hydroxyapatite for bone tissue regeneration.
Future perspective
handheld printer

• The most advanced 3D printing application that is
anticipated is the bioprinting of complex organs. It has
been estimated that we are less than 20 years from a
fully functioning printable heart.
• Although, due to challenges in printing vascular
networks, the reality of printed organs is still some way
off, the progress that has been made is promising. As the
technology advances, it is expected that complex
heterogeneous tissues, such as liver and kidney tissues,
will be fabricated successfully

1. Guillemot F, Mironov V and Nakamura M, Bioprinting is coming of age: report from the International
Conference on Bioprinting and Biofabrication in Bordeaux (3B'09). Biofabrication, vol.2(1): 010201 (2010).
2. Saunders, R. E. & Derby, B. Inkjet printing biomaterials for tissue engineering: bioprinting. Int. Mater. Rev.
59, 430–448 (2014).
3. Norotte, C., Marga, F. S., Niklason, L. E., and Forgacs, G. Scaffold-free vascular tissue engineering using
bioprinting. Biomaterials 30, 5910–5917 (2009).
4. Karve, S. S., Pradhan, S., Ward, D. V. & Weiss, A. A. Intestinal organoids model human responses to
infection by commensal and Shiga toxin producing Escherichia coli. PLoS ONE 12, e0178966 (2017).
5. Burdick, J. A., and Prestwich, G. D. Hyaluronic acid hydrogels for biomedical applications. Adv. Mater.
Weinheim 23, H41–H56. doi:10.1002/ adma.201003963 (2011).
6. Ortega-Prieto, A. M. et al. 3D microfluidic liver cultures as a physiological preclinical tool for hepatitis B
virus infection. Nat. Commun. 9, 682 (2018).
Reference

3D BIOPRINTING: PRINCIPLE, TECHNIQUES AND IT’S APPLICATION IN HUMAN THERAPY

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