Artificial Organs

Tissue Engineering
Dr. Pragasam Viswanathan
Professor, SBST

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Ð Tissue engineeing is closely associated with
applications that repair or replace portions of or whole
tissues i.e.
 Bone
 cartilage
 blood vessels
 bladder
 Skin

Ð Tissuesrequired certain mechanical & structural

properties for proper functioning.

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 In 1987 - Tissue engineering was coined at a
National Science Foundation (N.S.F.)
bioengineering meeting in Washington D.C

 VACANTI & LANGER,

―A combination of the principles & methods of
life sciences with that of engineering, to develop
materials & methods to repair damaged or diseased
tissues, & to create entire tissue replacements
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NEED FOR TISSUE ENGINEERING

 Tissue engineering holds promise of producing

better organs for transplant. Using tissue
engineering techniques & gene therapy it may be
possible to correct many otherwise incurable genetic
defects.
 A major goal of tissue engineering is in-vitro
construction of transplantable vital tissue.
 Artificial tissues can revolutionize healthcare by
providing a supply of soft & hard CT on demand.
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TRIAD

CELLS

Time

REGENERATED
TISSUE
Appropriate
Environment

SCAFFOLDS SIGNALLING

MOLECULES

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 Successful tissue engineering requires interplay
among three components:

 Implanted & cultured cells that will create

new tissue;

 Biomaterial to act as scaffold or matrix to

hold cells;

 Biological signaling molecules that instruct

cells to form desired tissue type.
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STRATEGIES TO ENGINEER TISSUES

 Characterised in 3 major classes

 Conductive

 Inductive

 Cell transplantation approaches

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TECHNIQUES

2 methods
 In Vitro
 In vivo

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 IN VITRO

 Construction in laboratory of vital tissue & its

subsequent implantation into host body.

 Advantage is ability to examine tissues as

they are formed, & to perform specific tissue
measurements.

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 By in-vitro TE of tissues such as bone, need
for recruitment of specific cells to site is
negotiated & predictability of regeneration is
enhanced,

overcoming many of limitations with

conventional therapies.

 Disadvantage is absence of a physiologic

environment
 Implanted tissue has to be incorporated with
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 IN VIVO

 Indicates obvious advantage of tissue regeneration in-

vivo in which incorporation occurs as tissues are
formed.
 This has formed basis for tissue engineering,
 which now includes implantation of porous matrices, seeded
with appropriate cells & signalling molecules, to facilitate
tissue regeneration in-vivo.

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 Disadvantage of in-vivo approach

regenerating tissues may get dislodged or

degraded by mechanical forces acting normally
at site, beforeregenerated tissue is fully formed &
incorporated

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CELL
S
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SOURCES

 Autologous cells (the host’s own cells)

 Allogenic cells (cells from a donor)

 Xenogenic cells (cells from a different species)

 Stem cells: either allogenic (fetal or adult

derived) or autologous (adult derived).

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Autologous cells are obtained from same
individual to which they will be re-implanted.
Have fewest problems with rejection &
pathogen transmission, however in some cases
might not be available.
Example genetic disease suitable
autologous cells are not available.
These cells can differentiate into a
variety of tissue types, including
bone, cartilage, fat, & nerve.
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Allogeneic cells come
from body of a donor of
same species.
Employment of dermal
fibroblasts from human
foreskin has been
demonstrated to be
immunologically safe &
thus a viable choice for
TE of skin.
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Xenogenic cells are
these isolated from
individuals of another
species. In particular
animal cells have been
used quite extensively in
experiments aimed at
construction of CV
implants.

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STEM CELLS
 Undifferentiated cells with ability to divide in culture &
give rise to different forms of specialized cells.

 Characteristic Features:
 They are capable of dividing & renewing themselves for
long periods
 They are unspecialized
 They can give rise to specialized cell types.

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 Stem cells could be:
 Adult stem cells
 Embryonic stem cells

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APPLICATION

Cell Replacement Therapies

Cells could be stimulated to develop into specialized
cells that represent renewable sources of cells &
tissue for transplantation.

Cell replacement therapy could treat injuries &
various genetic & degenerative conditions including
muscular dystrophies, retinal degeneration,
Alzheimer disease, Parkinson's disease, arthritis,
diabetes, spinal cord injuries, & blood disorders such
as hemophilia.

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SCAFFOLD
S
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SCAFFOLDS
 Used to
 guide
 organization,
 Growth & differentiation of cells in process of forming
functional tissue
 provide both physical & chemical signals.
 Tissues are composed of
 cells,
 insoluble extracellular matrix (E.C.M.)
 soluble molecules that serve as regulators
of cell function.

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 E.C.M. usually composed of 3 components:
 Collagen

 Glycoprotein

 Proteoglycan

 The E.C.M. is important for

 Growth

 Function - various cell types involved.

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TYPES OF SCAFFOLDS

ABSORBABLE NON-ABSORBABLE

SYNTHETIC NATURAL
POLYMERS POLYMERS
•P.L.A. •Collagen
•P.G.A •Fibrin
•Chitosan

NATURAL SYNTHETIC SYNTHETIC

MINERALS POLYMERS CEREMICS
•organic Bone •Polytetra •Calcium
flouroethylene Phosphate

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SIGNALLING
MOLECULE
S
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SIGNALLING MOLECULES

 Signalling molecules or biologic modifiers -

materials or proteins & factors that have
potential to alter key cellular events in host
tissue, by stimulating or regulating the wound
healing process.

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MODE OF ACTION

SYSTEMIC
(ENDOCRINE) LOCAL
(PTH,GH,LH)

JUXTACRINE
PARACRINE AUTOCRINE INTRACRINE
(Stem cell
(PDGF, TGF-β) (BMPs, TGF-α) (PTHrp)
factor)

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GROWTH AND DIFFERENTIATION FACTORS

 Growth factors - play important role in

regeneration are:
1) Platelet derived growth factor
(P.D.G.F.),
2) Insulin-like growth factor (I.G.F.),
3) Transforming Growth Factor- β (T.G.F.-
β),
4) Fibroblast Growth Factor,
5) Bone Morphogenetic Proteins (B.M.P.s).

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 PRODUCTION: Several cell types
produce PDGF, including
 Degranulating platelets,
 Smooth muscle cells,
 Fibroblasts,
 Endothelial cells,
 Macrophages & keratinocytes.

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RECOMBINANT BMP-2 PRODUCTION

 Recombinant proteins are produced from one of

several cellular expression systems:
 Bacteria,

 Insect cells or mammalian cells.

 rh BMP-2 is produced using mammalian cell

expression system, which allows for proficient
execution of post-translational modifications that
are present in human BMPs.

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 Chinese Hamster Ovary (CHO) cells are host of
choice. Because mammalian cells synthesize a
variety of GF, they are capable of synthesizing &
secreting active BMP.

 Includes many steps:

 Synthesizing of precursor polypeptide chains.
 Correct refolding & demineralization of these chains,
 Glycosylation of protein.

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INSULIN-LIKE GROWTH FACTORS (IGF-I,II):

 Peptide growth factors

with biochemical &
functional similarities
to insulin.
 Bone cells produce &
respond to IGF’s, and
bone is a storage house
for these factors in their
inactive form.

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TRANSFORMING GROWTH FACTOR-β:

 Multifactorial growth factor, structurally related to

B.M.P.s, but functionally quite different.
 Chemotactic for bone cells, & may increase or decrease
their proliferation depending upon the differentiation
state of the cells, culture conditions and concentration of
TGF-β applied.
 In-vivo, produces new cartilage and / or bone, if injected
in proximity to bone; however, it does not induce new
bone formation when implanted
away from a bony site.

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BONE MORPHOGENETIC PROTEINS (BMPS):

 Urist in 1965, reported that protein extracts

from bone, implanted into animals at non-
bone sites induced formation of new cartilage
& bone tissue.

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FIBROBLAST GROWTH
FACTORS:
 Family of at least 9 related gene products of which 2
major members are a-FGF or FGF-1 & b-FGF or FGF-
2.
 Stimulate endothelial cells & PDL cell migration &
proliferation, as well as stimulation of bone cell
replication. ell

 b-FGF is more potent than a-FGF & may act via

stimulation of other growth factors like TGF-β.

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PLATELET RICH PLASMA
 PDGF & TGF-β are well-established wound healing
―hormones

 One of highest concentrations of PDGF & TGF-β in

body are found within α-granules of blood platelets

 Thus, concentrating platelets would result in

concentration of growth factors, enhancing wound
healing on application.

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 Promotes angiogenesis.

 Haemostatic properties.

 Dense fibrin net that is highly osteoconductive.

 High concentrations of leukocytes, which act as

―autologous antibiotic, reducing risk of infection.

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 Future developments in fields of molecular & cell
biology, developmental biology & tissue engineering,
will have significant impact on managing anatomic
changes due to disease process.

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THANK
YOU
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