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MSE 598/494 Bio-inspired Materials and Biomaterials MSE 598/494 Bio-inspired Materials and Biomaterials
Instructor: Ximin He
TA: Xiying Chen Email: xchen128@asu.edu
2014-04-08
Lecture 13-14. Tissue Engineering
Organs, Cell Seeding and TE scaffolds
Artificial lung
Organs
Artificial heart
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Tissues
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blood vessel regeneration
Bone repair Bone healing
Tooth Bone/Enamel reconstruction
cardiac tissues engineering
Cell Seeding & Scaffold
cartilage formation Porous alginate scaffold
artificial skin growth
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Tissue engineering
Themes:
Generating functional organs (or organ components) outside of the body,
e.g. on the laboratory bench-top, which could be subsequently implanted
and/or substituted for damaged or defective organs.
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Constructing functional tissues for
testing of drug candidates and
examination of biological processes and
phenomena in environments which
best mimic actual physiological and
cellular conditions
Fundamental requirement of
organ/tissue created in an artificial environment
To morphologically resemble its native counterpart
To perform similar biological functions
Tissue engineering
Definition: The structural and functional reconstitution of tissues in
which the cells, biomaterials, and biological signals are combined and
fully mimic their physiological settings.
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Challenges: Engineered tissues
need to
be physiologically compatible, i.e.
integrate well and satisfactorily
perform their functions after
implantation into the body.
respond and transform external
stimuli in the same way as native
tissues for example lengthening
and thickening of a muscle tissue
following extended physical exercises
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What you will learn in the next 90 minutes?
Tissue Engineering:
Organ Tissue Cell
Lecture 13. Artificial Organs
Artificial Kidney
Artificial lung, heart (option for Lit Rev Presentation)
Lecture 14. Cell Seeding and TE Scaffold
Porous (polysaccharides, hydrogel, printing, carbon
microstructures)
Responsive & Dynamic (4-D scaffold)
Tissue engineering and drug design (option for Lit Rev
Presentation)
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MSE 598/494 Bio-inspired Materials and Biomaterials MSE 598/494 Bio-inspired Materials and Biomaterials
Instructor: Ximin He
TA: Xiying Chen Email: xchen128@asu.edu
2014-04-08
Lecture 13. Tissue Engineering
Artificial Organs
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Organs from bench
yet huge challenges
Complex structure with huge number of
cells
Organs from others
find donors
immunosuppressant drugs to prevent
rejection
Organs grown from patients own cells
incapable of regeneration slow
Creating body parts for damaged or defective organs
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Artificial Organs
Strategies:
partial organ regeneration
regeneration of organ constituents, ultimately embedding them as
substitutes for damaged or defective counterparts within the native organ.
to use a hierarchical approach in which core elements of the organ are
synthetically constructed, while more peripheral parts are produced in
more conventional manners.
Challenges of incorporation of synthetic, foreign assemblies in the
human body:
the embedded component has to adhere to specific requirements in terms
of functionality, long-term stability, and robustness.
more crucial and often problematic is the issue of host rejection of the
foreign object
Current Research: the relatively simple and already available as products
(commercial production of human skin for burn therapy) OR growing heart,
kidney, liver and similarly complex organs
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Engineered kidney membranes
Creating artificial renal membrane-like layers constitute a platform for
seeding and growth of kidney epithelial cells on one side, endothelial
cells (for regeneration of blood vessels) on the other side
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Epithelial side
Endothelial side
Dankers, P.Y.W. et al., From kidney development to drug delivery and tissue engineering strategies
in renal regenerative medicine, J. Controlled Rel. 2011 152, 177185. Copyright (2011)
Artificial cell layers for kidney-tissue engineering Macro-scale organization of the
polymer scaffold for cell growth
100 nm
To induce growth of functional cell monolayers on scaffold:
Appropriate morphology & biocompatibility
Appropriate physical/chemical properties:
mechanical flexibility, high surface area, and permeability
Human organ implantation in mice
Technique for creating artificial liver, implanting the organ in mice, and
using the humanized mice for biological and pharmacological studies.
Primary hepatocytes are cultivated on a polymer scaffold together with
liver endothelial cells, creating an artificial liver that is subsequently
implanted in a mouse (i.e. ectopic liver).
The humanized mouse model enables examination of varied drug
effects and biological processes associated with the implanted liver.
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Chena, A. A. et al., PNAS 2011 108, 1184211847.
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Levels of Creating Artificial Organs
Varies Challenges
1. Type of tissue being recreated in the laboratory
2. Properties of the scaffold materials used, esp. stability in the
physiological environment and biocompatibility
3. Ultimately the viability and integration of the produced tissues
in their target locations in the body
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Recreation of entire organs Organ
Constructing artificial tissues in lab
Subsequently implanting them in the
body
Tissue
growth and proliferation of healthy
and functional cells outside of body
Cell
MSE 598/494 Bio-inspired Materials and Biomaterials MSE 598/494 Bio-inspired Materials and Biomaterials
Instructor: Ximin He
TA: Xiying Chen Email: xchen128@asu.edu
2014-04-08
Lecture 13. Tissue Engineering
Cell Seeding & TE Scaffold
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Example - Cardiac Tissues
Requirement for impact and benefits for Heart diseases or those
experiencing myocardial infarction or heart failure:
distinct mechanical, electrical, and metabolic properties of heart tissue which
are ultimately responsible for efficient systolic and diastolic performance
Ideal artificially-created cardiac tissue: mimic the functional and
morphological characteristics of a native heart muscle:
capable of efficient contraction and expansion,
respond to electrophysiological signals,
exhibit long-term mechanical stability,
develop an effective vascular (i.e. blood vessel) network after implantation
not be rejected by the body
Three components of TE research
growth and proliferation of healthy and functional cells
proper design of scaffolds upon which the cells will assemble
achievement of effective biological signaling within the artificially
created tissue
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Challenges in cell seeding and growth in engineered tissues:
Limited availability of desired cells which often had to be carefully matched to
the individual in whom the tissue will be grafted in order to minimize tissue
rejection (stem cell technology)
The risk of adversely affecting cell properties during cell growth in engineered
tissues
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Tissue regeneration through seeding cells in blood vessels
Discovery against conventional knowledge: stem cells
embedded and grown within laboratory-engineered
blood vessels gradually disappeared after implanting the
vessels around the heart
Seeded cells can recruit host immune cells to the
engineered tissue for subsequent rapid formation of new
blood vessels
Employ standard cell lines Trigger the immune
system to induce vascularization within the artificial
tissue Enable growth of the endogenous cells
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1wk 6 wk 10 wk
Roh, J .D. et al., PNAS 2010 107, 46694674.
TE scaffolds
Focus of TE:
Facilitating appropriate scaffold
configurations in order to recreate the
actual organization and
environments of the tissues within
the body
Controlling the microscopic and
macroscopic architecture of the
scaffold
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TE scaffold materials
A critical precondition for effective tissue growth is the need for the
scaffold material to be both
biocompatible and biodegradable
Identify new materials to exhibit specific physical properties and
architectures allowing their use in TE designs
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Biological tissue Artificial tissue
Porous tissue scaffoldings for tissue
regeneration for cardiac repair
copoly(ether-esters)-polyamides blend hydrogel
(synthetic hydrogel)
Biological tissue scaffold revealed a porous
structure with an apparent interconnectivity
Collagen
hydrogel
(Natural
hydrogel)
10 m
6 m
Bone scaffold
Collagen hydrogel-ceramic composite
(natural-synthetic blend)
1 cm
1) Porous TE Scaffolds
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1) Porous TE Scaffolds
Synthetically Controlled Structures and Surface Chemistry
host cells in their pore structure.
large internal surface areas for cell attachment, proliferation, and physical
support for the tissue formed
efficient transport of molecules to and from cells (vital metabolic pathways,
signaling, and cell-cell communication)
Chung, H.J . and Park, T.G., Surface engineered and drug releasing pre-fabricated scaffolds for tissue
engineering, Adv. Drug Deliv. Rev. 2007 59, 249262.
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1) Porous TE Scaffolds - materials
Materials:
poly lactide glycolic acid (PLGA)
polysaccharides
chitosan
cellulose
alginate
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Porous alginate scaffold
Advantages of Porous Scaffold:
Interconnecting networks of pores for
effective migration
growth and attachment of cells
transport of nutrients and cell secretions
Mechanical stability and inert framework materials unreactive with
cells, while biodegradable through digestion by proteolytic enzymes
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i) Hydrogels
Functions:
1. Serving as scaffold:
mimics the physiological (hydrous) conditions in the human body
biological macromolecules, particularly proteins
allow exploiting the protein gel framework to act both as the scaffolding,
means to generate biological signals to the attached cells
2. Three-dimensional models for cell migration and proliferation
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Endothelial biomaterials
Vascular endothelial growth factor
(VEGF), within the porous
framework of hydrogels designed
to facilitate spatial guidance of cell
growth, particularly endothelial cells
(ECs) lining blood vessel walls
VEGF-mimic peptide fibers for
blood vessel regeneration
the VEGF-mimic peptide assembles
into nanofibers; showing the porous
gel structure formed by the peptide
fibers
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ii) Rapid prototyping (RP)
solid free form (SFF) fabrication and aided by advances in three-
dimensional printing are capable of producing complex rigid
structures directly from computer models
Especially for bone
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Fedorovich, N.E. Organ printing:thefuture of bone regeneration,
Trends Biotech. 2011 29, 601606.
accomplish cell deposition and seeding
concurrent with template fabrication
iii) Artificial skin
Inclusion of the natural protein
collagen in a polysaccharide
polymer scaffold is essential for
promoting cell attachment and
proliferation within the
scaffold.
Only with collagen, skin
fibroblast cells succeed to form
a continuous graft
physical organization of the
collagen fiber network,
to putative biological cues
provided by this protein
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polymer scaffold:
poly-DL-lactic-co-glycolic acid (PLGA) mesh
Cell proliferation:
only PLGA
PLGA+collagen
30 mins 5 days
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iv) Wood
remarkable resemblance of the microchannel structure in Rattan wood to
human bone
withstanding heavy loads, microscopic stretching, and shape flexibility
Unique hierarchical microstructural architecture of wood confers an
attractive combination of high strength,stiffness and toughness at relatively
low material density.
Sprio, S. et al., Biomimesis and biomorphic transformations: New concepts applied to bone
regeneration, J. Biotech. 2011 156, 347355.
v) Carbon nanostructures
Alignment of cells on an electrically-stimulated carbon nanotube
scaffold. Effects of a carbon nanotube scaffold and electrical
stimulation (duration: 14 days) upon the morphology of seeded
mesenchymal stem cells (MSCs).
Pronounced
elongation and cell
alignment were clearly
linked to application
of electrical
stimulation through
the carbon nanotube
cell-growth scaffold
Mooney, E. et al., The electrical stimulation of carbon nanotubes
to provide a cardiomimetic cue to MSCs, Biomaterials 2012 33, 61326139.
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vi) Endothelial biomaterials
Synthetic vascular grafts
gradually coated with
endothelial cells (ECs)
Vascularization (blood
vessel formation) of
endothelial scaffolds
Modular tissue engineering:
Small scaffolds pre-seeded with
ECs can be implanted directly at
the target site, or delivered in a
larger container.
The ECs form an interconnected
network, ultimately producing a
viable EC layer enabling blood
transport.
Mooney, E. et al., The electrical stimulation of carbon nanotubes to provide a cardiomimetic cue to MSCs,
Trends Biotech. 2012 26, 61326139.
at the implanted site
at the implanted site Pre-implantation
Pre-implantation
Diffuse Dense cytoskeleton
Apoptosis Proliferation
Stiffness
Size
Howcellsbehaveinmechanically dynamic environment?
CellMechanobiology, Mechanotransduction,Cytoskeletaldynamics
Cell culture materials
Petri dish Natural ECM Artificial ECM
2D semi-3D 3D (3D+time) 4D?
Static Dynamic
Chris Chan et al, Nature 2011
Ansethet al, Science, 2009
React to cellular mechanical and chemical signals
Spatio-temporally tunable
strain
Gel: homogeneous@nano
cellular response to gel structure
& mechanics:
heterogeneous@micro
Mechanics spatio-temporally
Assaying Stem Cell Mechanobiologyon Microfabricated
Elastomeric Substrates with Geometrically Modulated Rigidity
2) Responsive Dynamic TE scaffolds
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Dynamic scaffolds to study cell biology in four dimensions
Spread & move divide cell: Round
Bond open Pore size increases gel: Bond close
1. Interaction 2. In-situ monitor
3. Color mapping 4. Damage sensing
New Features: tunable in space and time
by 1) external or 2) cell-induced stimuli
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Concept:
Using responsive gel with active crosslinkers
Cellularly/Enzymatic degradable peptide linker
Isomerizable linker (reversible)
Steps:
1. Creating responsiveness to mechano, along with
other signals, such light and pH, with color change
2. Chemical cues
Advantages:
Initiate chemreaction with mechanical force:
Provide precise spatial and temporal control over
1) Bond formation; 2) Degradation
3) Pendant ligand tethering and releasing
Cell ECM
dictate/
regulate
direct
remold
respond
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Hydrogels with Enzymatic Cleavage of Networks
ECM-mimicking hydrogels composed of PEG-based hydrogel
crosslinked by the oligopeptides which are cleavable by the
matrix metalloproteinases (MMPs).
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Summary
Tissue Engineering:
Organ Tissue Cell
Lecture 13. Artificial Organs
Artificial Kidney
Artificial lung, heart (option for Lit Rev Presentation)
Lecture 14. Cell Seeding and TE Scaffold
Porous (polysaccharides, hydrogel, printing, carbon
microstructures)
Responsive & Dynamic (4-D scaffold)
Tissue engineering and drug design (option for Lit Rev
Presentation)
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Reading Resources
Prof. David Mooney (hydrogel based scaffold)
Prof. Kit Parker (cardiac cell based jellyfish)
4-D cell culturing scaffold
Ear on mice
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Homework of Lecture 13-14
1. Please state the general requirements for creating tissue
engineering scaffold, using an example of successful scaffold
materials.
Due by 04/17/2014
Hand in hard copy of homework at the TA, Xiying Chen, at the
beginning of the 04/17 class
Please contact xchen128@asu.edu for questions.