Tissue

Engineering
Dr. Siti Sunarintyas

Definition
Tissue Engineering : the in vitro development of tissues
or organs to replace the function of injured body parts.

Research on several different

types of tissues and organs :
Skin
Blood Vessels
Bone
Muscle
Nerves
Liver
Kidney
etc.

How it works ??
Three basic steps in tissue engineering :
1. getting the base cells to work with
2. putting the altered cells into a scaffold in order
to incubate the cells
3. putting the newly created cells or organ into use.

Step 1- Creating the Materials

The materials that are used for tissue
engineering are actually living cells that
were taken from somewhere else.
These cells are then modified to whatever
kind of cell is needed.
There are 7 sources from where you can get
these cells.

7 Sources of Cells
Autologous- from the
person himself.
Allogenic- from a body with
the same species.
Xenogenic- from a different
species.
Syngenic- from genetically
identical people.(Twins).
Primary- from any
organism.
Secondary- from a cell
bank.
Stem cells- Undifferentiated
cells.

Step 2-Bioreactors and Scaffolds

Bioreactors are used to create physiological
environments so that the cells can grow in a
natural manner.
Once the cells are obtained, then put into a
scaffold to let them grow into the new forms.
The problem of creating new organs : they
would die before they placed into the organism.
This problem has been solved with the
developments in 3D printers, which allows for
blood vessels to be put into the new organ.

8 Types of Scaffolds
Nano fiber SelfAssembly
Textiles
Solvent Casting &
Particulate Leaching
Gas Foaming
Emulsification/Freezedrying
Thermally Induced
Phase Separation
Electrospinning
CAD/CAM

Step 3- Usage
This step obviously is when the newly created
cells are put into use.
Whether they are being used to create a new
organ via the 3D Organ Printer, or they are
being used as new skin.

Future
Tissue engineering is a very promising field in
Biomedical engineering.
It can solve many problems that people
experience today.
It will eventually continue to grow and make
the need for a donor list obsolete as they will
be able to just grow organs specifically for
people.

Tissue Organization
Before a tissue can be developed in vitro, we
must understand how tissues are organized.
The basic tenant is :

all tissues are comprised of

several levels of structural hierarchy
These structural levels exist from the
macroscopic level (centimeter range) to the
molecular level (nanometer range)

Organization of the Tendon

Organization of the Kidney

Cellular Communications
Soluble Signals:

small proteins (15-20 kDa) which are chemically stable with long halflives (unless specifically degraded)
growth factors, steroids, hormones, cytokines, chemokines
bind to membrane receptors usually with high affinity (low binding
constants: 10-100 pM)
can diffusion long distances

Cell-to-Cell Contact:
some membrane receptors are adhesive molecules
adherent junctions and desmosomes
other serve to create junctions between adjacent
cells allowing for direct cytoplasmic communication
gap junctions
1.5-2 nm diameter and only allow transport of
small molecules ~1 kDa

Cell-ECM Interactions:
ECM is multifunctional and also provides a substrate that cells can

communicate
since cells synthesize the ECM, they can modify the ECM to elicit specific
cellular responses
several specialized receptors that allow for cell-ECM interactions
integrins, CD44, etc.
also a mechanism by with cells respond to external stimuli (mechanical
transducers)

Tissue Engineering Scaffolds

Biomaterial Scaffolds Materials:
Polymeric : chitosan, alginate, hydrogels, fibres
Natural : collagen, elastin, fibrin
Ceramic : calcium phosphate based for bone tissue
engineering
permanent versus resorbable
degradation typically by hydrolysis
must match degradation rate with tissue growth
Chemical and Physical Modifications:
attachment of growth factors, binding sites for
integrins, etc.
nanoscale physical features

Culturing of Cells
Types of Cell Culture
monolayer (adherent cells)
suspension (non-adherent cells)
three-dimensional (scaffolds or templates)

Sterilization Methods
ultra-violet light, 70% ethanol, steam autoclave,
gamma irradiation, ethylene oxide gas
Growth Conditions
simulate physiological environment
pH 7.4, 37C, 5% CO2, 95% relative humidity
culture (growth) media replenished periodically
Culture (Growth) Media
appropriate chemical environment
pH, osmolality, ionic strength, buffering agents
appropriate nutritional environment
nutrients, amino acids, vitamins, minerals,
growth factors, etc.

Cell Sources
primary cells
tissue biopsy, low cellular yield , potential age-related
problems
passaged cells
serial expansion of primary cells (can increase
population by 100-1000X)
tendency to either lose potency or de-differentiate with
too many passages
stem cells
undifferentiated cells
self-renewal capability (unlimited?)
can differentiate into functional cell types
very rare

Stem Cells
Stem cells naturally exist in some tissues (especially
those that rapidly proliferate or remodel) and are
present in the circulation.

There are two predominant lineages of stem cells:

mesenchymal
give rise to connective tissues (bone, cartilage,
etc.)
although found in some tissues, typically
isolated from bone marrow
hematopoietic
give rise to blood cells and lymphocytes
isolated from bone marrow, blood (umbilical
cord)

Stem cells are rare; bone marrow typically has:

a single mesenchymal stem cell for every
1,000,000 myeloid cells
a single hematopoietic stem cell for every 100,000
myeloid cells

Stem Cells (Mesenchymal)

Stem Cells (Hematopoietic)

Colony-Forming Units (CFUs)

Bioreactors
a) Spinner Flask:

semi-controlled fluid shear

can produce turbulent eddies which
could be detrimental

b) Rotating Wall

low shear stresses, high mass transfer

rate
can balance forces to stimulate zero
gravity

c) Hollow Fibre

used to enhance mass transfer during

the culture of highly metabolic cells

d) Perfusion

media flows directly through construct

e) Controlled Mechanics

to apply physiological forces during

culture

Tissue model
construction

Tissueengineeringrequiresthreethings:

Cells

Scaffold

Signals

 The scaffold refers to the matrix within the

tissue model construct
The signals refer to molecular signaling
molecules, also known as growth factors

Basic scaffold criteria:

Portions need to be biodegradable
Usually designed in the shape of the tissue
product the researcher is working on