Biofabrication and 3D-Bioprinting

Lecture 1
Introduction to Biofabrication and 3D-Bioprinting

Prof. Dr.-Ing. Andreas Blaeser

Institute for BioMedical Printing Technology

IDD
IDD
 Agenda for today
Short overview on the general topics of this lecture

• Presentation of the IDD

• Cell culture essentials
• Biofabrication methods
– Tissue engineering methods
– Bioassembly methods
• 3D-bioprinting
– Formulation and characterization of bioinks
– Bioprinting methods
– Influence of shear stress on living cells
– Bioprinting strategies
• Applications
– Bioprinted implants
– Bioprinted in-vitro models
– Non-medical applications
• Summary and organizational details
https://www.engineering.com/3DPrinting/3DPrintingArticles/ArticleID/6688/3
D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 2
 Goals of the lecture
The students should be able…

• To explain and differentiate the basic biofabrication

technologies and the bioprinting process chain.
• To compare different biofabrication technologies,
naming advantages and disadvantages for special
applications.
• To be able to analyze calculate the physical
interactions (e.g. shear stress) and phenomena of
fluid transport in individual 3D bioprinting
processes.
• To explain the essential elements and components
of a 3D-bioprinting system.
• To be able to name exemplary applications of
biofabrication and 3D-bioprinting technologies.
• To propose different biofabrication technologies for
specific medical and non-medical applications.
• To be able to make predictions on the applicability
and prospects of success of special biofabrication
processes and 3D bioprinting methods with regard
https://www.engineering.com/3DPrinting/3DPrintingArticles/ArticleID/6688/3
to the cell types and target tissues to be processed. D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 3
 Agenda for today
Short overview on the general topics of this lecture

• Presentation of the IDD

• Cell culture essentials
• Biofabrication methods
– Tissue engineering methods
– Bioassembly methods
• 3D-bioprinting
– Formulation and characterization of bioinks
– Bioprinting methods
– Influence of shear stress on living cells
– Bioprinting strategies
• Applications
– Bioprinted implants
– Bioprinted in-vitro models
– Non-medical applications
• Summary and organizational details
https://www.engineering.com/3DPrinting/3DPrintingArticles/ArticleID/6688/3
D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 4
IDD - Institut für Druckmaschinen und Druckverfahren

• The institute was founded in 1953 by seven

printing machine manufacturers.
• Today the institute comprises two
departments within the facutly of mechanical
engineering with approximately 20 coworkers
and 14 scientists: Prof. Dr.-Ing. Andreas Blaeser Prof. Dr.-Ing. Edgar Dörsam

– Prof. Dr.-Ing. Andreas Blaeser

BioMedical Printing Technology
– Prof. Dr.-Ing. Edgar Dörsam
Printing Science and Technology
• Infrastructure: 2D- and 3D-printing lab, 3D-
bioprinting lab, KUKA robotics platform,
climate room, S1-cell culture lab,
characterization microscopy laboratory, color
measurement lab, technical center,
mechanical workshop.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 5
Research focus of Prof. Dörsam

• Automation and measurement technology

– Development and automation of new processes and the measurement of
process parameters.
– Validation of physical models or creation of empirical models through large
data analyses.
– Application of National Instruments platform and software LabVIEW, MATLAB
and Python as well as AI and machine learning.

• Printed Functionality
– Printing processes for the use of new functional materials and the realization
of new, innovative applications (e.g. printed electronics, biosensorics).
– Through upscaling processes fundamental knowledge can be transferred into
technology.

• 3D printing (additive manufacturing)

– Research focuses on filament-based technologies (FDM, FFF) for the creation
of fine porous structures (e.g. filters, bones) and on the decoration of 3D
surfaces.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 6
Research focus of Prof. Blaeser

• Biofabrication and 3D-bioprinting

– Research and development of 3D-bioprinting systems,
automated cell assembly methods and bioreactors.
– Modelling and experimental investigation of different
mechanisms and phenomena for the transport of fluid
and solid biomaterials and their interaction with living Bild: 3D-Biodrucker (Blaeser et al., 2016).

cells.
• Biomedical applications
– Bioprinted Organ-on-a-Chip systems for
pharmacological and toxicologial screenings and
research. Bild: Biogedrucktes Gewebemodell (Blaeser, et
al. 2016).
– Bioprinted implants for regenerative medicine.
• Non-medical applications
– Bioprinted consumables (e.g. food, leather).
– Bioprinted sensors, actors, and bio-devices.
Bild: Mikrofluidchip-Entwicklung.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 7
 Agenda for today
Short overview on the general topics of this lecture

• Presentation of the IDD

• Cell culture essentials
• Biofabrication methods
– Tissue engineering methods
– Bioassembly methods
• 3D-bioprinting
– Formulation and characterization of bioinks
– Bioprinting methods
– Influence of shear stress on living cells
– Bioprinting strategies
• Applications
– Bioprinted implants
– Bioprinted in-vitro models
– Non-medical applications
• Summary and organizational details
https://www.engineering.com/3DPrinting/3DPrintingArticles/ArticleID/6688/3
D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 8
Cell culture essentials

Theoretical part
• What is a cell?
• What are its primary functions?
– Adhesion
– Proliferation
– Migration
https://ecampusontario.pressbooks.pub/microbio/chapter/uniqu – Differentiation
e-characteristics-of-eukaryotic-cells/

Practical part
• How do we culture cells?
• What are the main
components of a cell lab?
• How do we analyze cells?

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 9
 Agenda for today
Short overview on the general topics of this lecture

• Presentation of the IDD

• Cell culture essentials
• Biofabrication methods
– Tissue engineering methods
– Bioassembly methods
• 3D-bioprinting
– Formulation and characterization of bioinks
– Bioprinting methods
– Influence of shear stress on living cells
– Bioprinting strategies
• Applications
– Bioprinted implants
– Bioprinted in-vitro models
– Non-medical applications
• Summary and organizational details
https://www.engineering.com/3DPrinting/3DPrintingArticles/ArticleID/6688/3
D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 10
 Source: Medizin und Technik (https://medizin-und-
technik.industrie.de/medizintechnik-studium/faszination-medizintechnik/tissue-
engineering-lebende-ersatzteile-kommen-aus-dem-labor)

IDD
Tissue engineering

Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser

TISSUE ENGINEERING
24.04.2024
11
Tissue engineering

Cells Scaffolds
TE
Biological
factors

Source: Operation Karriere (https://www.operation-

karriere.de/karriereweg/assistenzarzt/ueberblick-weiterbildung-anatomie.html)

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 12
Excusrion: aortic prosthesis and bypass surgery

Material compatibility

Textile grafts, e.g.:

• PTFE (Gore-Tex®),
• PET (Dacron®).
Aortic arch / descendes replacement

Donor site morbidity

• Redirect blood around
blocked artery
• Healthy blood vessel
from leg, arm or chest
Coronary bypass surgery Source: Universitätsklinik für Herzchirurgie LKH Innsbruck
(http://herzchirurgie.uki.at/page.cfm?vpath=aufgabengebiete/thorakale_aorta); https://www.cardio-guide.com/therapie/bypass-op/;

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 13
Tissue engineering methods

– Hydrogel casting
– Winding "cell carpets" around a core
– Cell seeding on preformed scaffold

Cell-hydrogel Winded Tubular scaffold

suspension “cell carpet”

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 14
 Tissue engineering methods

Cell-hydrogel Winded Tubular scaffold

suspension “cell carpet”

C. B. Weinberg and E. Bell, 1986 N. L’Heureux et al., 1998 L. E. Niklason et al., 1999

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 15
Tissue engineering methods

Cell-hydrogel Winded Tubular scaffold

suspension “cell carpet”

• Disadvantages
– Moulds and cores are needed and must be adapted as required
– Geometric freedom (branching, etc.) extremely restricted
– Limited in fabricating complex multimaterial / multi-cellular structures
(required for more complex organs)
IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 16
Tissue engineering methods
Scaffold based TE methods

Lyophilization

Textile Salt leaching /

technology solvent casting

Sponge- / foam-
replication

3D-printing Decellularization

FR/SR: Montazeri et al., 2015 TT: Johnstone et al., 2013

3DP: Huang et al., 2018 LP: Lin et al., 2004
SL/SC: Yu et al., 2016 DC: Baiocchini et al., 2016

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 17
Tissue engineering methods
Example: tissue engineering of human bladder

Source: Nature (https://www.nature.com/news/2006/060403/full/060403-3.html)

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 18
Tissue engineering methods
Example: tissue engineering of human bladder

• In 1991 Antonthy Atala implanted

the first tissue engineered human
bladder
• The tissue was generated using
manual cultivation steps
• Labor extensive and difficult to
reproduce
Source: BBC (https://www.bbc.com/news/business-45470799)

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 19
 Agenda for today
Short overview on the general topics of this lecture

• Presentation of the IDD

• Cell culture essentials
• Biofabrication methods
– Tissue engineering methods
– Bioassembly methods
• 3D-bioprinting
– Formulation and characterization of bioinks
– Bioprinting methods
– Influence of shear stress on living cells
– Bioprinting strategies
• Applications
– Bioprinted implants
– Bioprinted in-vitro models
– Non-medical applications
• Summary and organizational details
https://www.engineering.com/3DPrinting/3DPrintingArticles/ArticleID/6688/3
D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 20
 What is bioprinting about?
https://www.engineering.com/3DPrinting/3DPrintingArti
cles/ArticleID/6688/3D-Printing-Organs-with-the-
Bioficial-Technique.aspx

Robotics, Microfluidics, Biology, Chemistry, Software, 3D-imaging

Control Engineerging Biomaterials • Data acquisition
• 3-axis robotics • Cell function • Data processing
• Nanoliter dispensing • Bioink development (segmentation, print
• Controlling the printing • Nutrient supply, cell and path analysis)
parameters (T, p, V, …) tissue maturation • User interface
IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 21
 Why do we need 3D-BIOPRINTING?

ECONOMICAL CONSIDERATIONS
TECHNICAL CONSIDERATIONS
• No need for molds or cores
• Geometrical freedom
• Multi-material /-cell structures

• Scalability
• Quality
3D-BIOPRINTING
IDD • Standardization
Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 22
Three phases of bioprinting

3D-design Tissue model

Bioink

Reifung
Maturation
-- Dyn.
Dyn. cultivation
Kultivierung
3D-Druckprozess
3D-printing process - Angiogenesis
-- Dispenser - Angiogenese
Dispensiereinheit - Implantation
- Robotics - Implantation
-- Controller
Robotik
Vorbereitung
Preparation - Steuerung
-- 3D-Design
3D-design
-- Materialsynthese
Material synthesis
-- Zellzüchtung
Cell culture Blaeser (2016) Dissertation

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 23
Challenges

• Preparation
– Biomimetic 3D-modelling
– Data processing
– Intelligent algorithms for print path
calculation and vasculature distribution

• Printing process Kinstlinger and Miller (2016), Lab on a Chip

– Dispensing cells without damage

– Selection of biomaterials
– Different printing strategies
– Technical aspects (precision, sterility,
reproducibility)

• Maturation
– Nutrient supply of macroscopic cell constructs
– Dynamic cultivation in bioreactors
– Online monitoring of the maturing process
(biological readouts) natgeotv.com

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 24
Printing methods

3D-
Bioprinting
Printing Drop
Layer Line Spheroid
entity

Stereo- Micro- Laser- Micro-

Technology Inkjet Acoustic
lithography extrusion based valve

Process
0
illustration

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 25
 Printing methods

3D-
Bioprinting

Print format Layer Line Droplet

Photo- Micro- Laser- Micro-

Technique Inkjet
patterning extrusion based valve

Process
illustration

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 26
 Photopatterning Microextrusion
Mirror Force F Pressure P Torque T
Printing methods
Plunger
Connector Extrusion
screw
Syringe Bioink
Projector/ light source

Photopolymer 3D-
Moveable Bioprinting
plattform Syringe pump Pneumatic Extruder

Print format Layer Kolesky et al. (2016),

LineAdv. Mater.
Droplet

Photo- Micro- Laser- Micro-

Technique Inkjet
patterning extrusion based valve

Process
illustration

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 27
 Laser-induced forward transfer (LIFT)

Unterschiedliche
Pulsed laser
Drucktechniken
Glass slide (Donor)
beam

Absorbance layer

Shock wave Hydrogel-cell

Jet suspension
3D-
Glass slide
(Collector) Bioprinting

Print format Layer Line Droplet

Photo- Micro- Laser- Micro-

Technique Inkjet
patterning extrusion based valve

Process
illustration

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 28
 Inkjet printing
Thermal inkjet Piezo-electric inkjet
Unterschiedliche Drucktechniken
Piezoelement
Heating
element Membrane

Vapor
3D- bubbles
Fluid- Nozzle
Bioprinting
Bioink reservoir opening

Wilson and Boland, 2003.

Print format Layer Line Droplet

Photo- Micro- Laser- Micro-

Technique Inkjet
patterning extrusion based valve

Process
illustration

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 29
 Microvalve based bioprinting
Pressurized air
Unterschiedliche Drucktechniken
Stationary
anchor
Fluid
reservoir Switch
Coil
Microvalve
Mobile anchor
Spring
3D-
Bioink drop
Valve ball Bioprinting
Valve seat

Print format Layer Line Droplet

100
80
Viability / %

60
Photo-
Viscosity Process Micro- Laser- Micro-
Technique
40 Inkjet
patterning
Nozzle geometry extrusion based valve
20 control
Static pressure
0
0 5 10 15 20
Shear stress / kPa

IDD
Blaeser et al. (2016), Adv. Healthc. Mater.
Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 30
 3D-
Bioprinting

Print format Layer Line Droplet

Photo- Micro- Laser- Micro-

Technique Inkjet
patterning extrusion based valve

Process
illustration

Macroscopic
objects

Blaeser et al. (2017) Curr. Ma et al. (2016), Kang et al. (2016), Yan et al. (2013), Christensen et al. (2015), Blaeser et al. (2016),
Opin. Biomed. Eng. Proc. Natl. Acad. Sci. Nat. Biotechnol. Biofabrication Biotechnol. Bioeng. Adv. Healthc. Mater.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 31
 3D-
Bioprinting

Print format Layer Line Droplet

Photo- Micro- Laser- Micro-

Technique Inkjet
patterning extrusion based valve

Process
illustration

Microscopic
structures

Blaeser et al. (2017) Curr. Ma et al. (2016), Kolesky et al. (2016), Guillotin et al. (2010) , Cui et al. (2009), Lee et al. (2009),
Opin. Biomed. Eng. Proc. Natl. Acad. Sci. Proc. Natl. Acad. Sci. Biomaterials Biomaterials Biomaterials

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 32
What is a hydrogel?
Consumables

Wound dressings

Contact lenses

3D-cell culture

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 33
 WS Lecture: Biomaterials
What is a hydrogel? and Tissue Engineering

Challenging
Water swollen polymer network
3D printing process
• Visco-elastic behavior
• High water content (> 90 %)
• Low polymer content (0.1-3.0 %)
• ECM-like properties
• Ideal environment for cell growth
• Low mechanical properties

Blaeser et al. (2016), Topuz et al. (2018) 3D-cell culture

https://www.igb.fraunhofer.de/en/research/functional-surfaces-and-materials/biomaterials/services-and-equipment.html

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 34
 Heat resistance (°C)
Excursion: the hydrogel dilemma 250
PEEK
PPS
200 Hydrogels
150
PP PC PA
100 ABS
1900-2700PEMPas
ABS PVC PLA
New 3D-Printing 50
Strategies
Shape Fidelity

0 50 100
Tensile strenght (MPa)
Traditional
Biofabrication
Window Collagenous
Blood Brain Muscle Bone
Cell Culture
Fluid 1 kPa 10 kPa 100 kPa

10 % Serum
MSC Collagen I

Mechanics h Substrate elasticity (E)

& Rheology
Malda et al. (2013) Advanced Materials

Two Challenges Engler et al. (2006) Cell

(1) Low mechanical properties  Poor shape fidelity

IDDsensitivity
(2) High biological 
Introduction to Biofabrication andSmall variations
3D-Bioprinting have
| Prof. Dr.-Ing. big Blaeser
Andreas influence
24.04.2024 35
 Hydrogel
Example of our own work in this fieldblending
Hydrogel functionalization (Agarose-Collagen)
Tissue Eng. Part A 2015
Biofabrication 2016
Adv. Healthc. Mater. 2016
Tissue Eng. Part C 2017

Microstructural modification α-helix

(Carboxylated Agarose)
Dr. Aurelien Forget
(University of South Australia)
Prof. Prasad Shastri β-sheet
(University of Freiburg)
Adv. Healthc. Mater. 2017
Biofabrication 2017

Nanoparticle integration
(Iron-loaded Alginate)
Prof. Stephan Barcikowski
(University Duisburg-Essen)
Nano Res. 2016

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 36
Example of our own work in this field
Biofunctionality
Hydrogel functionalization
Tissue Eng. Part A 2015
Mechanics Biofabrication 2016
Adv. Healthc. Mater. 2016
Tissue Eng. Part C 2017

Microstructural
Mechanics modification α-helix
(Carboxylated Agarose)
Rheology
β-sheet

Biofunctionality
Nanoparticle integration
(Iron-loaded Alginate)
Mechanics

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 37
 Example of our own work in this field
Hydrogel blending

COL0.36
AG0.5-COL0.21
Collagen ↑ AG1
Agarose ↓ AG1-COL0.10
AG2-COL0.05

0 50 100
Blood Brain Muscle Collag. Bone 100 µm
E-Modulus (kPa)
Fluid 1 kPa 10 kPa 100 kPa
Stiffness
10 % Serum
MSC Collagen I

Angiogenesis in bulk material

h Substrate elasticity (E) hMSC & HUVEC Coculture

Engler et al. (2006) Cell; Duarte Campos, Blaeser et al. (2016) Adv
Healthc Mater, IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 38
 The Hydrogel “Dilemma”

New 3D-Printing
Solutions
Strategies
- New hydrogel formulations
Shape Fidelity

Traditional
- New printing strategies
Biofabrication
Window

Cell Culture Hybrid printing

Core-shell
Submerged structures

Shell material
Mechanics
& Rheology Core material

Malda et al. (2013) Advanced Materials Blaeser et al.al.

(2013) Bioresearch,
Akkineni
Shim et al.et(2011)
(2016) Biofabrication
Biofabrication
Duarte Campos et al. (2013) Biofabrication
Two Challenges
(1) Low mechanical properties  Poor shape fidelity
IDDsensitivity
(2) High biological 
Introduction to Biofabrication andSmall variations
3D-Bioprinting have
| Prof. Dr.-Ing. big Blaeser
Andreas influence
24.04.2024 39
 Example of our own work in this field
Different bioprinting methods and strategies
Submerged Bioprinting Biofabrication 2013
Biores. Open Access 2013
Patent WO 2013 113883 A1

Bioextrusion

RSC Adv. 2014

Nano Res. 2016 1cm
Biomacromolecules 2016

Contactless Bioprinting

IDD Adv. Healthc. Mater. 2016

Adv. Healthc.
Introduction to Biofabrication Mater. 2017
and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 40
 Example of our own work in this field
Drop-on-demand 3D-bioprinting systemc

Air pressure 3D-Bioprinter

3-axis robotic
• Additive manufacturing
of cell-laden hydrogel
Temperature- structures
sensor
Heating element • Four heatable and
independently
Cell-gel
controllable printer
suspension heads

Microvalve
• Microvalves enable
precise deposition of
Printing platform droplets in nanoliter
range

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 41
 Example of our own work in this field
Drop-on-demand 3D-bioprinting system

Air pressure
5 mm
3-axis robotic

Temperature-
sensor
Heating element
5 mm

Cell-gel
suspension

Microvalve

500 µm
Printing platform

Blaeser et al. (2016) Adv Healthc Mater

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 42
 Example of our own work in this field
Hybrid bioprinting strategy

Until now:
The drop constitutes the basic
building block in DoD-bioprinting
Mechanically
stable Bioink Biofunctionalization

Novel approach:
Mechanical reinforcement Biofunctional
Using the interface between
Bioink
drops to guide capillary growth

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 43
 Agenda for today
Short overview on the general topics of this lecture

• Presentation of the IDD

• Cell culture essentials
• Biofabrication methods
– Tissue engineering methods
– Bioassembly methods
• 3D-bioprinting
– Formulation and characterization of bioinks
– Bioprinting methods
– Influence of shear stress on living cells
– Bioprinting strategies
• Applications
– Bioprinted implants
– Bioprinted in-vitro models
– Non-medical applications
• Summary and organizational details
https://www.engineering.com/3DPrinting/3DPrintingArticles/ArticleID/6688/3
D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 44
 Medical goals of bioprinting
I Printing & cultivation
II Evaluation
III Clinical trials
Animal trials IV Patient care
Implantation

Tissue substitutes
1
Patient

Bioprinting

Drug screening

In-vitro models
2 Therapy
2003 2017 Blaeser et al. (2017) Curr. Opin. Biomed. Eng.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 45
 Bioprinted tissue implants
I Printing & cultivation
II Evaluation
III Clinical trials
Animal trials IV Patient care
Implantation

Tissue substitutes
1
Patient

Bioprinting

Drug screening

In-vitro models
2 Therapy
2003 2017 Blaeser et al. (2017) Curr. Opin. Biomed. Eng.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 46
 Bioprinting tissue implants

A B C

3D CAD model Medical Imaging

Tissue
Bioprinting product
Blaeser et al. (2017) Curr. Opin. Biomed. Eng.

IDD Kang et al. (2016), Nature Mater.

Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 47
 Bioprinting tissue implants
Examples Material Cell types In-vivo-assessment Culture
Bone RGD-Alginate, Murine MSC Bone volume / area after 12 weeks
Daly (2016) PEGMA, GelMA, PCL implantation (µCT imaging) (mouse)
Duarte Agarose, Collagen hMSC n. a. 3 weeks
Campos (2016) (in vitro)
Cui (2016) PLA, GelMA hMSC, n. a. 4 weeks
HUVECs (in vitro)
Gao (2015) PEGDA, GelMA, hMSC n. a. 3 weeks
(in vitro)
Kang (2016) PCL, Gelatin, HA, Human AFSCs Bone / osteoid formation, blood 5 months
Fibrinogen, Glycerol vessel formation (rat)
Cartilage Alginate Chondrocytes n. a. 3 week
Yu (2016) (in vitro)
Abbadessa PEG, polyHPMA-lac, Chondrocytes n. a. 6 weeks
(2016) CSMA, HAMA (in vitro)
Costantini GelMA, CS-AEMA, hMSC n. a. 3 weeks
(2016) HAMA, Alginate (in vitro)
Gao (2016) PEGDA hMSC n. a. 3 weeks
(mouse)
Kang (2016) PCL, Gelatin, HA, Rabbit ear Ear cartilage reconstruction, Blood 2 months
Fibrinogen, Glycerol chondrocytes vessel formation (mouse)
Muscle tissue TPU, PCL, gelatin, Myoblasts, n. a. 1 week
Merceron (15) fibrinogen, HA fibroblasts (in vitro)
Kang (2016) PCL, Gelatin, HA, Myoblasts Muscle function (electromyo- 2 weeks
Fibrinogen, Glycerol graphy), Innervating capability (rat)
Neural tissue Polyurethane Murine NSCs Rescue function after traumatic 10 days
Hsieh (2016) dispersions brain injury (locomotion, survival) (zebra fish)
Osteochondral PCL, Atelocollagen, hTMSC Evaluation of neocartilage by ICRS 8 weeks
Shim (2016) HA scoring system (rabbit)
Pancreas PCL, Alginate HUVECs, Islets Glucose induced insulin secretion 4 days
Marchioli (16) of Langerhans, test (CAM assay)
Sweat glands Gelatin, alginate murine EPCs Functional restoration of sweat 2 weeks
Huang (2016) glands in mice (mouse)
Blaeser et al. (2017)

IDD Curr. Opin. Biomed. Eng.

Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 48
 Bioprinting tissue implants

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024
Source: 49 (2019)
Lee et al., Science
 Bioprinting tissue implants

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024
Source: 50 (2019)
Lee et al., Science
 Bioprinted in-vitro models
I Printing & cultivation
II Evaluation
III Clinical trials
Animal trials IV Patient care
Implantation

Tissue substitutes
1
Patient

Bioprinting

Drug screening

In-vitro models
2 Therapy
2003 2017 Blaeser et al. (2017) Curr. Opin. Biomed. Eng.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 51
 Bioprinting in vitro models
E G

H I

F
Blaeser et al. (2017) Curr. Opin. Biomed. Eng.

IDD Homan (2016), Sci. Report.

Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 52
 Bioprinting in vitro models
Examples Material Cell types Clinical readout Culture
Alveolar tissue Matrigel Endothelial and n. a. 5 days
Horváth (2016) epithelial cells)
Bone tissue PLA, Gelatin, hMSC, HUVECs n. a. 28 days
Cui (2016) Polylysine
Brain tumor Gelatin, Fibrin Glioma stem cells, Tumor sensitivity to the chemot- 21 days
Dai (2016) Alginate Human glioma cells herapeutic Temozolomide
Breast cancer GelMA + nHA BrCa cells, fetal n. a. 14 days
Zhou (2016) osteoblasts, hMSC
Cardiac tissue GelMA, HUVECs, Influence of Doxorubicin on beating 15 days
Zhang (2016) Alginate cardiomyocytes rate and vWF Level
Lind (2016) Dextran, TPU, NRVMs, hiPS-CMs Influence of Verapamil and 28 days
ABS, PDMS, Isoproterenol on the contraction force
Ag:PA, PLA (inotropic response)
Hepatic tissue HA, Gelatine, hepatocytes, stellate Albumin and urea production 20 days
Skardal (2015) PEG cells, kupffer cells
Ma (2016) GelMA Hepatic progenitor n. a. 20 days
cells, ASCs, HUVECs
Lee (2016) PCL, gelatin, HepG2, HUVEC Albumin and urea production 6 days
collagen
Nguyen (2016) NovoGel® 2.0 human hepatocytes, Influence of Trovafloxacin and Levo- 28 days
stellate cells, HUVECs floxacin on LDA, albumin, and ATP
Neural tissue Alginate, hNSCs Bicuculline-induced increased calcium 10 days
Gu (2016) Agarose, CMC response
Renal tissue Gelatin, Fibrin, PTE cells, fibroblasts, Influence of Cyclosporine A on 42 days
Homan (2016) Pluronic immunecells epithelial viability and barrier function
Skin tissue Collagen Dermal Fibroblasts Transdermal penetration ability of 7 days
Hou (2016) nanoparticles
Thrombosis GelMA Fibroblasts, HUVECs Thrombolysis by tPA flow 14 days
Zhang (2016 II)
Blaeser et al. (2017) Curr. Opin. Biomed. Eng.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 53
 Agenda for today
Short overview on the general topics of this lecture

• Presentation of the IDD

• Cell culture essentials
• Biofabrication methods
– Tissue engineering methods
– Bioassembly methods
• 3D-bioprinting
– Formulation and characterization of bioinks
– Bioprinting methods
– Influence of shear stress on living cells
– Bioprinting strategies
• Applications
– Bioprinted implants
– Bioprinted in-vitro models
– Non-medical applications
• Summary and organizational details
https://www.engineering.com/3DPrinting/3DPrintingArticles/ArticleID/6688/3
D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 54
Summary & Outlook

• Following more than 10 years of research the

printing technology is already quite advanced

• Remaining challenges
– Biomaterials / Bioinks
– Biofunctionalization / tissue maturation

• Promising approaches, BUT

– Scale
– Mechanics
– Cultivation of multiple cell types

• In-Vitro models
– Commercial applications just started
– Bioprinted organoids in cosmetic and pharma
industry (skin, leather, kidney, heart, etc.)

• Tissue implants
– Simple printed tissue types in animal trials
(cartilage, bone)
– Parts of complex organs (heart, liver, kidney,
etc.) not within the next 5-10 years

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 55
 Organizational details
Topics of the upcoming lectures

• Cell culture essentials Lecture 2

• Biofabrication methods
– Tissue engineering methods Lecture 3-5
– Bioassembly methods

• 3D-bioprinting
– Formulation and characterization of bioinks
– Bioprinting methods Lecture 6-9
– Influence of shear stress on living cells
– Bioprinting strategies

• Applications
– Bioprinted implants
Lecture 10-11
– Bioprinted in-vitro models
– Non-medical applications

• Summary and final discussion Lecture 11 https://www.engineering.com/3DPrinting/3DPrintingArticles/Articl

eID/6688/3D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 56
Organizational details
Structure of the lectures  color coded slide titels

• Description of methods

• Example applications

• Examples of own work in the field

• Excursions

• Calculations and estimations

• Comparison of methods
https://www.engineering.com/3DPrinting/3DPrintingArticles/Articl
eID/6688/3D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 57
Organizational details
Information on the written exam

• The exam will be held as written exam

(60 min).

• The exact date and place of the exam

will be made public within the upcoming
weeks.

• In order to prepare for the exam a quiz

with exemplary questions will be
prepared and presented at the end of
some of the lectures.

https://www.engineering.com/3DPrinting/3DPrintingArticles/Articl
eID/6688/3D-Printing-Organs-with-the-Bioficial-Technique.aspx

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 58
Organizational details
Literature

• The slides of the presentation will be made

available one day before each lecture as
PDF.

• Besides the lecture slides, no addition

literature is required for the exam
preparation. However, for those interested
each chapter contains a list of literature for
further reading.

IDD Introduction to Biofabrication and 3D-Bioprinting | Prof. Dr.-Ing. Andreas Blaeser 24.04.2024 59
Thank you very much for your attention!

Technische Universität Darmstadt

Institute of BioMedical Printing Technology
Prof. Dr.-Ing. Andreas Blaeser
Magdalenenstr. 2
64289 Darmstadt
Germany

https://www.idd.tu-darmstadt.de

IDD