TISSUE

ENGINEERING

NAME: FRANKIE JONATHAN LALOO

ROLL NO:1378
M.Sc. (final)
INTRODUCTION
Tissue engineering evolved from the field of biomaterials
development and refers to the practice of combining scaffolds,
cells, and biologically active molecules into functional tissues.
The goal of tissue engineering is to assemble functional
constructs that restore, maintain, or improve damaged tissues
or whole organs. Artificial skin and cartilage are examples of
engineered tissues that have been approved by the FDA;
however, currently they have limited use in human patients.
HOW DOES IT WORK?
Cells are the building blocks of tissue, and tissues are the basic
unit of function in the body. Generally, groups of cells make and
secrete their own support structures, called extra-cellular
matrix. This matrix, or scaffold, does more than just support the
cells; it also acts as a relay station for various signaling molecules.
Thus, cells receive messages from many sources that become
available from the local environment. Each signal can start a
chain of responses that determine what happens to the cell. By
understanding how individual cells respond to signals, interact
with their environment, and organize into tissues and organisms,
researchers have been able to manipulate these processes to
mend damaged tissues or even create new ones.
The process often begins with building a scaffold from a wide set
of possible sources, from proteins to plastics. Once scaffolds are
created, cells with or without a “cocktail” of growth factors can
be introduced. If the environment is right, a tissue develops. In
some cases, the cells, scaffolds, and growth factors are all mixed
together at once, allowing the tissue to “self-assemble.”
Another method to create new tissue uses an existing scaffold.
The cells of a donor organ are stripped and the remaining
collagen scaffold is used to grow new tissue. This process has
been used to bioengineer heart, liver, lung, and kidney tissue.
This approach holds great promise for using scaffolding from
human tissue discarded during surgery and combining it with a
patient’s own cells to make customized organs that would not
be rejected by the immune system.

STEPS IN TISSUE ENGINEERING

Tissue engineering procedure involves several steps, which start

from cell selection, cell isolation, and culturing of primary
(progenitor or stem) cells; inducing their differentiation to
certain phenotypes; seeding and cultivation; design of adequate
scaffolds, including selection of proper materials and routes to
process, porosity, interconnectivity, surface characteristics, etc.
1) BIOPSY (donor-tissue extraction) - either from fluid tissue like
blood using centrifugation or apheresis (easier process) or from
solid tissue that involves more steps. Solid tissue is minced, then
enzymes like trypsin or collagenase are used to remove the
extracellular matrix, finally cells are free-floating and extracted
again by use of centrifugation or apheresis. Lately, trend in
donor-tissue extraction is emphasis on the non-invasive
methods.
2) CELL ISOLATION and CULTIVATION (manipulation with cells)
– it is safest to use autologous cells (primary cells extracted from
the same person's own healthy tissues to which the artificial
tissue will be transplanted), recently there has been trend to use
mesenchymal stem cells from bone marrow that can
differentiate into various tissue types. Other types of cells that
can be used are allogenic (donor from the same species),
heterologous (or xenogeneic, donor is different species). In
those cases, rejection of the host's immune system and possible
disease transmission are risks that need to be considered.

3) SCAFFOLDS, seeding, cultivation – implantation or 'seeding'

of cells into artificial structure that can support 3-D tissue
formation; that resemble the extracellular matrix. There are
certain requirements that scaffolds need to meet:
biocompatibility (acceptable for cells, high porosity and
adequate pore size), biodegradability and non-brittle nature (it
is preferable for scaffold to be absorbed by surrounding tissues),
can be functionalized with biomolecules, function as nutrition
for cells, adequate properties.
4) IMPLANTATION (implantation into living body)

5) DETECTION (property analysis)

 Figure 1: The classic tissue engineering (TE) paradigm
consists in the isolation of primary cell from the patient and
the further seeding on a three-dimensional (3D) porous
scaffold, specifically designed to induce cell proliferation
and differentiation. The bioartificial system is developed in
a specific environment, where the metabolic, mechanical
and electrical stimuli are provided by a bioreactor. In this
tissue-specific designed system, cells start to produce
extracellular matrix, leading to in vitro tissue maturation.
The tissue-engineered construct is then implanted in the
patient, undergoing to a remodeling process in vivo, which
allows the tissue restoration. (B) The TE approach for the
development of in vitro models. The first step is to
understand what part of the system considered should be
represented and which specific conditions should be met.
The second step is the design of the appropriate scaffold,
i.e., the design of a 3D porous construct recapitulating the
architecture and the mechanical and surface properties of
the tissue/organ to be modeled. The third step is the
selection of the cells to be seeded in the designed scaffold.
The subsequent steps should be the design and fabrication
of a bioreactor and the selection of the appropriate
chemical signals (e.g., cocktail of growth factors) to be
included in the model. Once a model is successfully
developed, it can be improved by increasing its complexity,
by introducing additional cell type, chemical factor or
coupling different mechanical stimuli. The final step is the
validation, aiming to demonstrate the relevance of the
model for the intended purpose. The validation procedure
is fundamental and this step should be carefully designed.

Assembly methods

Self-assembly – fabrication technique called micro masonry is

used to self-assemble micrometric and sub-micrometric 3D units
to larger structures - encapsulation of living cells in polymer
cubes.

Liquid-based template assembly - in this case an air-liquid

surface that is established by Faraday waves can be used as
template for formation of 3D network.
Additive manufacturing – 3D printing of models of vascular
system, organs. Successive layers of living cells are deposited on
gel medium or sugar matrix and then slowly built up to form 3D
structures – for example endothelial cells printed in sets of ring
and then incubated and fused into tubes. It is predicted that in
the future organs like kidneys and liver can be grown.
LIMITATIONS

Many complications are faced when developing artificial organs

using human tissue scaffolds. Many levels of sustainability are
impacted during the process. In terms of economics, human
tissue donors are very expensive, which makes it hard to obtain
resources and conduct consistent experiments. Additionally,
reliable crosslinking methods are way too costly.
Biocompatibility is also affected as seen in the need of cross-
linking agents. Decellularized human tissue scaffolds undergo
natural biodegradation processes, which drastically affects the
functionality of the final organ that’s created. This natural
breakdown of the artificial organ further disrupts vasculature
and may even produce harmful wastes. Lastly, because tissue
samples are donated, organ rejection could possibly occur when
the artificial organ undergoes in vivo testing. All these
shortcomings of human tissue scaffolds gave researchers the
incentive to seek out methods that would improve all the facets
of sustainability
References
 Wobma, Holly, and Gordana Vunjak-Novakovic. “Tissue
Engineering and Regenerative Medicine 2015: A Year in
Review.” Tissue engineering. Part B, Reviews vol. 22,2
(2016): 101-13. doi:10.1089/ten.TEB.2015.0535
 Caddeo S, Boffito M, Sartori S. Tissue Engineering
Approaches in the Design of Healthy and Pathological In
Vitro Tissue Models. Front Bioeng Biotechnol. 2017;5:40.
Published 2017 Jul 26. doi:10.3389/fbioe.2017.00040