An introduction course in Tissue Engineering for the people with mechanical/electrical engineering and physic scientists

1. Tissue Engineering
Introduction
September 2014
Ali Bakhshinejad
bakhshi3@uwm.edu

2. Tissue Engineering
“The application of the principles and methods of engineering and life sciences
toward the fundamental understanding of structure-function relationships in
normal and pathological mammalian tissue and the development of biological
substitutes to restore, maintain, or improve tissue function”
Skalak, R., & Fox, C. F. (1988). Tissue engineering: proceedings of a workshop, held at Granlibakken, Lake Tahoe,
California, February 26-29, 1988 (p. 343).

3. Tissue Engineering
They introduced a new field that is basically a combination
of engineering and biological science with motivation of
finding a solution for patients that suffer from organ failure.
Langer, R., & Vacanti, J. (1993, 01). Tissue engineering. Science, 260(5110), 920-926. doi: 10.1126/science.8493529

4. Motivation
The Need Is Real: Data. (n.d.). Retrieved June 24, 2014, from http://www.organdonor.gov/about/data.html
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5. Motivation
"Every 30 seconds, a patient dies from a disease that could
be treated with tissue replacement."
Dr. Anthony Atala
http://www.bioscaffold.com/
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6. Tissue Engineering is a combination of basic biological science, engineering
fundamentals, many clinical aspects and various relevant biotechnologies.
Required Backgrounds
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7. Tissue Engineering is a combination of basic biological science, engineering
fundamentals, many clinical aspects and various relevant biotechnologies.
- Biological Science:
Cell biology
Physiology
Embryology
Wound healing
Required Backgrounds
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8. - Clinical aspects:
Surgery and transplantation
Immunology
Pathology
Radiology
Medicine
Required Backgrounds
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9. - Biotechnologies:
Cell culture
Cell separation
Gene transfer
Required Backgrounds
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10. - Engineering fundamentals:
Fluid dynamics
Transport phenomena
Material Science
Mechanics
Chemical kinetics
Required Backgrounds
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11. - Engineering fundamentals:
Fluid dynamics
Transport phenomena
Material Science
Mechanics
Chemical kinetics
This long list make it challenging to cover all aspects in a course. This course
will be an introductory course with focus on Engineering fundamentals and few
important aspects of biotechnology.
Required Backgrounds
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12. Steps
Murphy, S. V, & Atala, A. (2014). 3D bioprinting of tissues and organs. Nature Biotechnology, 32(8), 773–
785. doi:10.1038/nbt.2958
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13. Steps
Murphy, S. V, & Atala, A. (2014). 3D bioprinting of tissues and organs. Nature Biotechnology, 32(8), 773–
785. doi:10.1038/nbt.2958
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14. Steps
Murphy, S. V, & Atala, A. (2014). 3D bioprinting of tissues and organs. Nature Biotechnology, 32(8), 773–
785. doi:10.1038/nbt.2958
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15. Steps
Murphy, S. V, & Atala, A. (2014). 3D bioprinting of tissues and organs. Nature Biotechnology, 32(8), 773–
785. doi:10.1038/nbt.2958
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16. Steps
Murphy, S. V, & Atala, A. (2014). 3D bioprinting of tissues and organs. Nature Biotechnology, 32(8), 773–
785. doi:10.1038/nbt.2958
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17. Steps
Murphy, S. V, & Atala, A. (2014). 3D bioprinting of tissues and organs. Nature Biotechnology, 32(8), 773–
785. doi:10.1038/nbt.2958
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18. Step 1: Imaging
X-Ray
X-rays are a type of electromagnetic radiation, just like visible light.
An x-ray machine sends individual x-ray particles through the body. The
images are recorded on a computer or film.
● Structures that are dense (such as bone) will block most of the x-ray
particles, and will appear white.
● Metal and contrast media (special dye used to highlight areas of the
body) will also appear white.
● Structures containing air will be black, and muscle, fat, and fluid will
appear as shades of gray.
http://www.nlm.nih.gov/medlineplus/ency/article/003337.htm
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19. Step 1: Imaging
X-Ray
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20. Step 1: Imaging
X-Ray
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21. Step 1: Imaging
Computed Tomography (CT)
Computed tomography (CT) is a type of imaging. It uses special x-ray
equipment to make cross-sectional pictures of subject.
CT scans are being used for
● Broken bones
● Cancers
● Blood clots
● Signs of heart disease
● Internal bleeding
● Tissue Engineering
http://www.nlm.nih.gov/medlineplus/ctscans.html
http://en.wikipedia.org/wiki/X-ray_computed_tomography
The Nobel Prize in Physiology or Medicine 1979 was awarded jointly to Allan M.
Cormack and Godfrey N. Hounsfield "for the development of computer assisted
tomography"
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22. Step 1: Imaging
Computed Tomography (CT)
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23. Step 1: Imaging
micro-Computed Tomography (micro-CT)
X-ray microtomography, like tomography and x-ray computed
tomography, uses x-rays to create cross-sections of a physical object
that can be used to recreate a virtual model (3D model) without
destroying the original object. The prefix micro- (symbol: µ) is used to
indicate that the pixel sizes of the cross-sections are in the micrometre
range. These pixel sizes have also resulted in the terms high-
resolution x-ray tomography, micro–computed tomography (micro-CT
or µCT), and similar terms. Sometimes the terms high-resolution CT
(HRCT) and micro-CT are differentiated, but in other cases the term
high-resolution micro-CT is used.Virtually all tomography today is
computed tomography.
Example:
http://upload.wikimedia.org/wikipedia/commons/d/d8/3D_rendering_of_
a_micro_CT_scan_of_a_piece_of_dried_leaf..ogg
http://en.wikipedia.org/wiki/X-ray_microtomography
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24. Step 1: Imaging
Magnetic Resonance Imaging (MRI):
An MRI (magnetic resonance imaging) scan is an imaging test that uses powerful magnets and radio waves to
create pictures of the body. It does not use radiation (x-rays).
Single MRI images are called slices. The images can be stored on a computer or printed on film. One exam
produces dozens or sometimes hundreds of images.
http://www.nlm.nih.gov/medlineplus/ency/article/003335.htm
http://en.wikipedia.org/wiki/Magnetic_resonance_imaging
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25. Step 1: Imaging
MRI VS CT:
Both MRI and CT scans have their own advantages and limitations.
But an important advantage of MRI is that no ionizing radiation is used and so it is recommended over
CT when either approach could yield the same diagnostic information
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26. Step 1: Imaging
Generating 3D geometry from 2D images
Baghaie, A., & Yu, Z. (2014). Curvature-Based Registration
for Slice Interpolation of Medical Images. In Y. Zhang & J. S.
Tavares (Eds.), Computational Modeling of Objects Presented
in Images. Fundamentals, Methods, and Applications SE - 7
(Vol. 8641, pp. 69–80). Springer International Publishing.
doi:10.1007/978-3-319-09994-1_7
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27. Step 2: Design Approach
First Idea was to print stem cells on
a bio scaffold presented by Langer,
R., & Vacanti, J. at 1993
Which is called Biomimicry method
Langer, R., & Vacanti, J. (1993, 01). Tissue engineering. Science, 260(5110), 920-
926. doi: 10.1126/science.8493529
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28. Step 2: Design Approach
Biomimicry method or Scaffold-base method
Derby, B. (2012, 12). Printing and Prototyping of Tissues and Scaffolds.Science,
338(6109), 921-926. doi: 10.1126/science.1226340
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29. Step 2: Design Approach
Biomimicry method or Scaffold-base method
Bioscaffolding is the use of biocompatible and bioresorbable
materials to construct a 3d structure comparable to the
implant tissue area, in order to promote tissue regeneration
and injury recovery. The structure is seeded with native
differentiable cells and cell adhesion proteins in order to
encourage cell adhesion and tissue regeneration. The matrix
is also consistently porous, which further promotes cell
adhesion and differentiation at a controlled rate. The scaffold
must be designed to withstand and effectively transfer local
stresses evenly across the area of implantation during the
degradation period. Also, the degradation properties are
catered to match the cell differentiation rates and
extracellular matrix deposition rates of the implant site in
order to provide continuous support throughout the repair
process. Materials used for the scaffold construction must be
chosen appropriately to minimize adverse reaction and
maximize cell adhesion and differentiation.
Derby, B. (2012, 12). Printing and Prototyping of Tissues and Scaffolds.Science,
338(6109), 921-926. doi: 10.1126/science.1226340
http://www.bioscaffold.com/
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30. Step 2: Design Approach
Biomimicry method or Scaffold-base method
http://www.nlm.nih.gov/medlineplus/ency/imagepages/1078.htm
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31. Step 2: Design Approach
Biomimicry method or Scaffold-base method
Gerlach, J. C., Johnen, C., Ottoman, C., Bräutigam, K.,
Plettig, J., Belfekroun, C., … Hartmann, B. (2011, April 1).
Method for autologous single skin cell isolation for
regenerative cell spray transplantation with non-cultured cells.
The International Journal of Artificial Organs. IJAO. Retrieved
from http://www.artificial-organs.com/article/method-for-
autologous-single-skin-cell-isolation-for-regenerative-cell-
spray-transplantation-with--non-cultured-cells-ijao-d-10-00181
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32. Step 2: Design Approach
Biomimicry method or Scaffold-base method
Ahn, S., Lee, H., Lee, E. J., & Kim, G. (2014, 12). A direct cell
printing supplemented with low-temperature processing
method for obtaining highly porous three-dimensional cell-
laden scaffolds. Journal of Materials Chemistry B. doi:
10.1039/c4tb00139g
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33. Step 2: Design Approach
“Could we direct write living cells?”
asked by primary investigators and
Defense Advanced Research Program Agency (DARPA)
program manager sometimes in the mid to late 1990s
Ringeisen, B. R., Othon, C. M., Barron, J. A., Young, D., & Spargo, B. J. (2006, 12). Jet-based methods to
print living cells. Biotechnology Journal, 1(9), 930-948. doi: 10.1002/biot.200600058
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34. Step 2: Design Approach
Self-assembly method
Jakab, K., Norotte, C., Marga, F., Murphy, K., Vunjak-Novakovic, G., &
Forgacs, G. (2010). Tissue engineering by self-assembly and bio-printing of
living cells. Biofabrication, 2(2), 022001. doi:10.1088/1758-
5082/2/2/022001
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35. Step 2: Design Approach
Self-assembly method
Prediction Models
Cellular Particle Dynamics (CPD)
Cellular Potts Model (CPM)
I. Kosztin, G. Vunjak-Novakovic, and G. Forgacs,
“Colloquium: Modeling the dynamics of multicellular systems:
Application to tissue engineering,” Rev. Mod. Phys., vol. 84,
no. 4, pp. 1791–1805, Dec. 2012.
F. Graner and J. Glazier, “Simulation of biological cell sorting
using a two-dimensional extended Potts model,” Phys. Rev.
Lett., vol. 69, no. 13, pp. 2013–2016, Sep. 1992.
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36. Step 2: Design Approach
Self-assembly method
Prediction Models Cellular Particle Dynamics (CPD)
I. Kosztin, G. Vunjak-Novakovic, and G. Forgacs,
“Colloquium: Modeling the dynamics of multicellular systems:
Application to tissue engineering,” Rev. Mod. Phys., vol. 84,
no. 4, pp. 1791–1805, Dec. 2012.
Mo RB St Im DA DC BP App Sy

37. Step 2: Design Approach
Self-assembly method
Prediction Models Cellular Particle Dynamics (CPD)
I. Kosztin, G. Vunjak-Novakovic, and G. Forgacs,
“Colloquium: Modeling the dynamics of multicellular systems:
Application to tissue engineering,” Rev. Mod. Phys., vol. 84,
no. 4, pp. 1791–1805, Dec. 2012.
r = position vector of the CP
= friction coefficient
F_1 = the force (due to intracellular interactions) exerted by
CPs inside the nth cell
F_2 = the force (due to intracellular interactions) exerted by
CPs in all the other cells in the system
f = is a manifestation of molecular fluctuations at the coarse-
grained CP level
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38. Step 2: Design Approach
Self-assembly method
Prediction Models Cellular Potts Model (CPM)
Thomas, G. L., Mironov, V., Nagy-Mehez, A., & Mombach, J.
C. M. (2014). Dynamics of cell aggregates fusion:
Experiments and simulations. Physica A: Statistical
Mechanics and Its Applications, 395, 247–254.
doi:10.1016/j.physa.2013.10.037
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39. Step 2: Design Approach
Self-assembly method
Prediction Models Cellular Potts Model (CPM)
F. Graner and J. Glazier, “Simulation of biological cell sorting
using a two-dimensional extended Potts model,” Phys. Rev.
Lett., vol. 69, no. 13, pp. 2013–2016, Sep. 1992.
type associated with cell
surface energy between spins
Lagrange multiplier
the area of the cell
A the target area for the cells of type
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40. Step 2: Design Approach
Cellular Potts Model (CPM)
F. Graner and J. Glazier, “Simulation of biological cell sorting
using a two-dimensional extended Potts model,” Phys. Rev.
Lett., vol. 69, no. 13, pp. 2013–2016, Sep. 1992.
Self-assembly method
Prediction Models
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41. Step 2: Design Approach
Mini-tissues
Mironov, V., Visconti, R. P., Kasyanov, V.,
Forgacs, G., Drake, C. J., & Markwald, R. R.
(2009). Organ printing: tissue spheroids as
building blocks. Biomaterials, 30(12), 2164–74.
doi:10.1016/j.biomaterials.2008.12.084
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42. Step 4: Cell Selection / Differentiated cells
In this method the differentiated cells will be selected for tissue engineering
purposes but here we will focus on cell differentiation process by it self
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43. Step 4: Cell Selection / Differentiated cells
Compartmental models
Continuous process
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44. Step 5: Bio-printing
Binder, K. W., Allen, A. J., Yoo, J. J., & Atala, A. (2011,
12). Drop-On-Demand Inkjet Bioprinting: A Primer.
Gene Therapy and Regulation, 06(01), 33. doi:
10.1142/S1568558611000258
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45. Step 5: Bio-printing
InkJet (Drop on Demand & continuous flow)
Faulkner-Jones, A., Greenhough, S., King, J. A., Gardner, J.,
Courtney, A., & Shu, W. (2013). Development of a valve-
based cell printer for the formation of human embryonic stem
cell spheroid aggregates. Biofabrication, 5(1), 015013.
doi:10.1088/1758-5082/5/1/015013
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46. Step 5: Bio-printing
InkJet (Drop on Demand & continuous flow)
Tasoglu, S., & Demirci, U. (2013, 12). Bioprinting for
stem cell research. Trends in Biotechnology, 31(1), 10-
19. doi: 10.1016/j.tibtech.2012.10.005
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47. Step 5: Bio-printing
InkJet (Drop on Demand)
Tasoglu, S., & Demirci, U. (2013, 12). Bioprinting for
stem cell research. Trends in Biotechnology, 31(1), 10-
19. doi: 10.1016/j.tibtech.2012.10.005
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48. Step 5: Bio-printing
Microextrusion (Continous flow)
Murphy, S. V, & Atala, A. (2014). 3D bioprinting of tissues and
organs. Nature Biotechnology, 32(8), 773–785.
doi:10.1038/nbt.2958
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49. Step 5: Bio-printing
Microextrusion (Continous flow)
Ozbolat, I. T., & Yu, Y. (2013, 12). Bioprinting Toward
Organ Fabrication: Challenges and Future Trends. IEEE
Transactions on Biomedical Engineering,60(3), 691-699. doi:
10.1109/TBME.2013.2243912
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50. Step 5: Bio-printing
Laser-assisted
Odde, D. J., & Renn, M. J. (2000, 12). Laser-guided direct
writing of living cells.Biotechnology & Bioengineering, 67(3),
312. doi: 10.1002/(SICI)1097-0290(20000205)67:33.3.CO;2-6
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51. Step 5: Bio-printing
Laser-assisted (LIFT)
Serra, P. (2006, 12). Laser-induced forward Transfer: A
Direct-writing Technique for Biosensors
Preparation.Journal of Laser Micro/Nanoengineering,1(3),
236-242. doi: 10.2961/jlmn.2006.03.0017
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52. Step 5: Bio-printing
Laser-assisted (modified-LIFT)
Guillemot, F., Guillotin, B., Fontaine, A., Ali, M., Catros,
S., Kériquel, V., ... Amédée-Vilamitjana, J. (2011, 12).
Laser-assisted bioprinting to deal with tissue
complexity in regenerative medicine. MRS Bulletin,
36(12), 1015-1019. doi: 10.1557/mrs.2011.272
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53. Step 5: Bio-printing
Laser-assisted
Guillemot, F., Guillotin, B., Fontaine, A., Ali, M., Catros, S.,
Kériquel, V., ... Amédée-Vilamitjana, J. (2011, 12). Laser-
assisted bioprinting to deal with tissue complexity in
regenerative medicine. MRS Bulletin, 36(12), 1015-1019.
doi: 10.1557/mrs.2011.272
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54. Step 6: Application
Application: Maturation
Cell Sorting
Jakab, K., Norotte, C., Marga, F., Murphy, K., Vunjak-
Novakovic, G., & Forgacs, G. (2010, 12). Tissue engineering
by self-assembly and bio-printing of living
cells.Biofabrication, 2(2), 022001. doi: 10.1088/1758-
5082/2/2/022001
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55. Step 6: Application
Application: Maturation
Tissue Fusion
Jakab, K., Norotte, C., Marga, F., Murphy, K., Vunjak-
Novakovic, G., & Forgacs, G. (2010, 12). Tissue engineering
by self-assembly and bio-printing of living
cells.Biofabrication, 2(2), 022001. doi: 10.1088/1758-
5082/2/2/022001
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56. Mo RB St Im DA DC BP App Sy
Syllabus
Week 2 X-Ray / CT / MRI
Week 3 Image processing /
Volume Rendering
Week 4 Quiz / Project 1
Week 5 Biomaterial Scaffolds
Week 6 CPD & CPM
Week 7 Quiz / Project 2
Week 8 Cell Differentiation *
Week 9 InkJet bio-printer
Week 10 Laser-assisted bio-printer
Week 11 Post-printing processes
Week 12 Final Project presentation
Class attendance 10%
Quiz 1/Project 1 20%
Quiz 2/Project 2 20%
Final Project 50%