CARDIOVASCULAR TISSUE ENGINEERING MODEL

Need of a Tissue Engineering model in Cardiovascular system

Cardiovascular diseases remain a signicant reason for morbidity and mortality in numerous
nations. Progress has been made in designing the different segments of the cardiovascular
framework, including veins, heart valves, and cardiovascular muscle. Moreover, advance
toward this objective will be made through the parallel improvement of successful tissue
designed parts of the cardiovascular framework that will later be combined into a bigger
organ structure, in particular a heart.

Biological and Technical aspects

Sources of Cells:

Type of cells most commonly used in cardiovascular tissue engineering:

Bone marrow-Derived mesenchyma stem cell

Multipotent adult germline stem cell
Induced pluripotent stem cell
Endogenous cardiac stem cell

Type of Culturing:

Hydrogel Technique
Prefabricated matrices
Decellularised heart tissue
Cell sheets

Type of developmental organogenesis formulation:

Organogenesis is the process of formation and development of the organs of an
organism from embryonic cells. During organogenesis, the three germ layers of the
embryo differentiate and further specialize to form the various organs of the body.
1. Endoderm : one of the three tissue layers in the embryo of a metazoan animal,
which will produce the digestive system and other internal organs of the adult.
2. Mesoderm : produces many internal organs of the adult such as the muscles, spine
and circulatory system.
3. Ectoderm : outermost layer which will produce the epidermis (skin) and nervous
system of the adult.

Biomaterial Technical Knowledge

Synthetic biomaterials:
 Synthetic materials are made in the laboratory from primary building blocks, so their
properties can be tuned to match desired characteristics.
They are free from animal origin.
Engineer can control the properties such as mechanical strength, biological activity,
degradation rates etc.
Examples:
i) Polyesters: polylactic acid (PLA), polyglycolic acid (PGA), poly (lactic-co-
glycolic acid) (PLGA).
ii) Polyurethanepoly(2-hydroxyethylmethacrylate) (pHEMA), poly-(N-(2-
hydroxypropyl) methacrylamide (pHPMA).
iii) Linear Polyether poly (ethylene oxide) (PEO).

Natural biomaterials:

Built-in structure, environment and cues similar to native body.

Examples- Fibrin, Hair keratin, Alginate, Hyaluronic acid, Collagen, Gelatin, ECM-
derived materials etc

Design criteria:

proper mechanical and physical properties.

adequate degradation rate without the production of toxic degradation products
suitable cell adhesion.
integration into surrounding tissue without extensive inflammatory response or
support of infection.
proper mass transfer.

CELL SIGNALING IN CARDIOVASCULAR TISSUE ENGINEERING MODEL

In Cardiac Tissue Engineering , the addition of exogenous electrical stimulation has
attributed to the reconstruction of an appropriate environment for tissue regeneration by
mimicking the pacemaker activity during heart tissue development.

JAK STAT SIGNALLING PATHWAY

 Scaffold Manufacturing Methods:
1. Solvent Casting and Particulate Leaching-
2. Freeze Drying
3. Gas Foaming
4. Electrospinning

Ethical Issues Related to Tissue engineering

The ethical and legal issues surrounding the use of human ESCas have obstructed further
exploration and shifted the existing attention onto other alternative sources for stem cell
therapy in cardiac repair.

PANCREAS AND INTESTINE TISSUE ENGINEERING MODEL

Need of Pancreatic and Instestinal Tissue Engineering model

Cell-based implantable artificial tissues are most promising for the long-term treatment of
endocrine diseases, such as diabetes.

. Recently, by utilizing principles gained from the study of developmental biology, human
pluripotent stem cells have been demonstrated to be capable of directed differentiation into
intestinal tissue in vitro..

Biological and Technical Aspects of Tissue Engineering of Pancreas and Intestine

The scaffolding material along with its cellular component must allow re epithelialization to
regenerate mucosal layers. Most importantly, the bioengineered tubular constructs must be
able to maintain a patent lumen and maintain integrity so as not to allow leakage during use
in the body as a replacement.

Biomaterial Technical Knowledge - SAME FOR ALL THE MODELS.

Biomaterials can be broadly classified into synthetic and natural.

A scaffold must possess the following properties:

Mechanical strength

Non-toxic to cells and surrounding tissues

Allow appropriate diffusion of nutrients and metabolites

Have sufficient mechanical integrity to withstand manipulations associated with

implantation and in vivo existence
 The degradation rate should mirror the rate of new tissue formation or be adequate for
controlled release of bioactive molecules.

Targets of Tissue Engineering Model

Two goals:

To improve insulin replacement therapy until glycemic control is practically normal

as evident by the avoidance of the complications of hyperglycemia.

To ease the burden of therapy for the insulin-dependent.

A biological approach to the artificial pancreas is to implant bioengineered tissue containing
islet cells. They secrete the amount on insulin, amylin, and glucagon needed in response to
sensed glucose.

Cell signaling

A central paradigm can be recognized in most events of cellular signaling, which consists of
three distinct steps:

Signal initiation : an extracellular ligand binds to a receptor on the surface of the cell.
Ligand binding changes activity of the receptor, thus generating the signal.

Signal transduction : The activated receptor triggers a signal transduction cascade in

which intracellular proteins are activated, ultimately leading to activation of a so-
called transcription factor in the cell s nucleus.

Gene activation : The transcription factor binds to regulatory sequences in target

genes, resulting in gene activation, protein synthesis and changed cellular physiology.

Signaling Pathways involved in intestinal and prancreatic stem cells:

Wnt pathway is the main driving force for the maintenance of the self-renewing
phenotype of the intestinal stem cells. Wnt signaling also induces the differentiation
of Paneth cells and regulates proper cell migration along the cryptvillus axis via the
cell-contact dependent Eph/Ephrin signaling.

The cell-contact dependent Notch pathway cooperates with Wnt to preserve the self-
renewal of the stem cells. Notch is also involved in the differentiation of absorptive
enterocytes vs. the secretory lineage through the lateral inhibition process.

BMP signaling is critical as BMPs support the differentiation of enteroendocrine cells

and the full maturation of the secretory cell lineage.

Hippo/YAP: the Hippo pathway is active in post-mitotic differentiated cells leading to

nuclear exclusion of YAP, suppressing proliferation.

Ethical Issues
Some ethical issues currently in tissue engineering are the use of stem cells, future patient
access, and patent claims.

BONE AND CARTILAGE TISSUE ENGINEERING MODEL

Bone and cartilage generation by autogenous cell/tissue transplantation is one of the most
promising techniques in orthopedic surgery and biomedical engineering.

NEED FOR BONE AND CARTILAGE TISSUE ENGINEERING - Madhur Malhotra

Engineered bone tissue has been viewed as a potential alternative to the conventional use of
bone grafts, due to their limitless supply and no disease transmission. However, bone tissue
engineering practices have not proceeded to clinical practice due to several limitations or
challenges. Bone tissue engineering aims to induce new functional bone regeneration via the
combination of biomaterials, cells, and factor therapy.

BIOLOGICAL AND TECHNICAL ASPECTS- Tulika Paul

BONE:
Bone tissue engineering (BTE) is based on understanding of bone structure, bone mechanics,
and tissue formation. Bone possesses ability to perform wide array functions and bone
responds to variety of metabolic, physical & endocrine stimuli.

BONE STRUCTURE & FUNCTION-

Bone contains three main bone-specific cell types-

Osteocyte.
Osteoblasts
Osteoclasts

CARTILAGE:

Cartilage is a tough, flexible connective tissue that is found in many areas of the body. This
fine, rubbery tissue mainly functions as a cushion for bones at joints.

There are three types of cartilage:

Elastic cartilage ( yellow cartilage) is the most springy and supple type of cartilage.
Fibrocartilage is the toughest type of cartilage,
Hyaline cartilage is both springy and tough.

CARTILAGE REPAIR AND REGENERATION:

Cartilage tissue engineering paradigm is based on the isolation of chondrocytes/ chondrocyte

precursors from a tissue biopsy, expanding the cell number in culture, seeding them onto 3D
scaffold, incubating for a period of time before placing the construct inside a patient.
Fig: General TE process for bone and cartilage model

BIOMATERIALS -Vipresha Jain

Scaffold

The scaffold is a 3-dimensional substrate and it serves as a template for tissues regeneration.
The ideal scaffolds should be porous and permeable to permit the ingress of cells and
nutrients..

Natural:

Collagen
Proteoglycans component glycosamigolycans and hyaluronan.
The adhesive proteins fibronectin and laminin
Hydroxyapatite

Synthetic Polymers:

poly(L-lactic acid) (PLA)

poly(glycolic acid) (PGA)
poly(D,L-lactic acid-co-glycolic acid) (PLGA)
poly(caprolactone)
poly (propylene fumarate)

Manufacturing Methods of Scaffolds - Asad Zuhaib

There are several methods for manufacturing a scaffold used in tissue culture. Some of them
are reviewed here in this section.

Solvent Casting

Particulate Leaching

ALTERNATIVE SOURCES FOR BONE AND CARTILAGE TISSUE

ENGINEERING
 Stem Cell Therapy:. Adult stem cells, such as mesenchymal cells, are commonly
found in the bone marrow and conjunctive tissues. They can produce certain cell
lines, such as osteoblasts, chondroblasts, adipocytes, tenocytes, myoblasts, and
stromal cells.
Autologous Bone Grafts: it represents a good source in terms of quality and quantity
of cancellous and cortical bone.
Regeneration with the help of inorganic materials: Ceramic materials are a group
of inorganic oxides and salts used in bone tissue engineering. Some ceramic materials
used in bone regeneration are listed below.

Calcium Phosphates: Calcium phosphates are a family of minerals composed of

calcium ions (Ca2+), orthophosphates (), metaphosphates or pyrophosphates (), and
sometimes hydrogen or hydroxide ions.

Bioglasses: Bioglasses are a family of bioactive glasses, compound of SiO2, Na2O,

CaO, and P2O5 in variable proportions.

CELL TARGETS AND CELL SIGNALLING Senjuti Roy Chowdhury

TGF Beta Signalling Pathway

 Fig: BMP SIGNALLING PATHWAY

ETHICAL CONCERNS Tamanna Maheshwari

Tissue engineering (TE) is a promising new field of medical technology. However, like other
new technologies, it is not free of ethical challenges. The issues that turn out to dominate the
debate are the use of human embryonic stem cells and therapeutic cloning. (Azizeh-Mitra
Yousefi, 2015).

There is an ethical concern over the use of Embryonic Stem Cells (ESCs) for bone and
cartilage tissue engineering. Although they present many advantages for regenerative
medicine and tissue engineering, many ethical issues limit their use and many countries
prohibit ESCs isolation from human embryo. (MARINA PANEK, 2015)

MARKETING

. To date, a large number of new bone and cartilage products and growth factor carrier
materials have been developed, but only few of them are approved for use in patients. Some
of the major reasons for the small numbers of approved products include, hurdles posed by
the cost of development, cost of goods, manufacturing scale-up, sterility and patent issues..
(Chhavi Sharma, 2011)

LIVER AND KIDNEY TISSUE ENGINEERING MODEL

BIOLOGICAL AND TECHNICAL ASPECTS OF LIVER AND KIDNEY TISSUE
ENGINEERING MODEL

1. SOURCES OF CELLS: Various cell types are being investigated as efficient sources of
hepatocytes for liver support system, cell transplantation and tissue engineering. Among them
are:

A) Primary human and porcine hepatocytes,

B) Tumor-derived and immortalized cell lines

C) Adult and embryonic stem cells

D) Fetal hepatoblasts

E) Oval cells and stem cell-derived hepatocyte-like cells

NEED OF TE MODEL

Why Tissue Engineering is necessary?

Producing better organs for implant

Use living cells as engineering material

Permanent implant

In-vitro construction of transplantable vital tissue

It save lives

Replace a structure with a completely living structure

Improve or replace tissues such as skin, muscle, bone

Improve or replace organs such as heart, liver, kidney

Eliminate risk of organ rejection

SCAFFOLDS AND ITS DESIGNING FOR TISSUE ENGINEERING

A number of three dimensional porous scaffolds have already been developed from various
kinds of biodegradable biomaterials.

Properties of scaffolds- 1: surface should promote cell adhesion, cell growth and allow
retention of differentiated cells. 2- The surface should be biocompatible implying neither the
polymer nor the bioproducts should invoke any antigenicity in the host. 3. The scaffold
should be degradable and eventually eliminated from the body and the material of the
scaffold should be mechanically strong.

ALTERNATIVES TO WHOLE ORGAN LIVER AND KIDNEY TISSUE ENGINEERING

Alternatives to whole organ liver and kidney tissue engineering are:

Bioartificial liver and kidney support systems- Bioartificial liver devices utilize
hepatocytes from animal or human sources. The bioartificial kidney has been used to treat
patients suffering acute renal failure and the results are promising.

Xenotransplantation- Xenotransplantation offers an unlimited supply of donor organs for

transplantation, and the majority of promising results have come from studies using porcine
organs.

TARGETS OF THE TISSUE ENGINEERING MODEL AND CELL SIGNALLING

TARGETS OF THE LIVER TISSUE ENGINEERED MODEL: Liver diseases, such as

infections, alcohol-related ailments, cancers, and autoimmune, metabolic, and drug induced
liver diseases represent a major health burden in modern times. Most of these conditions lead
to acute or chronic liver failure if no definitive treatment can be offered. The use of stem and
progenitor cells, liver derived, to be applied in the treatment of inherited and acquired liver
diseases could solve the problem of hepatocyte availability.

TARGETS OF THE KIDNEY TISSUE ENGINEERED MODEL: tissue-engineered kidneys

have been designed for the treatment of ischemic or toxic acute renal failure or acute tubular
necrosis and end-stage renal disease, which is currently treated through supportive therapies..

CELL-SIGNALLING:

During development, five ligandsreceptor signalling pathways are primarily employed:

TGF-, Wnt, RTK, Notch, and Hedgehog signalling pathways. These pathways all have a
role in triggering EMTs. Although the activation of a single-signalling pathway can be
sufficient for an EMT, in most cases an EMT or MET is initiated by multiple signalling
pathways acting in concert.

ETHICAL ISSUES- Same related to stem cells.

MARKETING OF LIVER AND KIDNEY TISSUE MODELS

Tissue engineering market size was valued at USD 5 billion in 2015 and is anticipated to
exhibit significant growth over the next seven years. Rising need to bridge the organ demand
and supply gap is expected to drive industry demand.