BME 6309 Tissue Engineering

Overview of Cell Biology,

Stem Cells, Cell differentiation,
Morphogenesis, Embryonic development, Tissue Homeostasis,
Cellular Signaling, Extracellular Matrix,
Natural Polymers in Tissue Engineering, Synthetic Polymers
in Tissue Engineering, Cytoskeleton and Cell Motility, Cell
Source and Cell Nutrition, Scaffold Design & Fabrication,
Bioreactors

2D Muscle Tissue Engineering , 3D Muscle Tissue

Engineering, Myogenesis, Contractility, Regenerative
Medicine,Controlled Release, Ophthalmic Tissue Engineering,
Cardiac Tissue Engineering, Current Topics in Tissue
Engineering, Future Perspectives Tissue Engineering.
Types of muscles in human body
Skeletal muscle tissue engineering
Z disc/ line, A band, I band, H
zone, M line
 Z disc/ line, A band, I band, H
zone, M line
Sarcomere: from one Z disc to
another Z disc
A band and I
band
 Steps of muscle
contraction
1. Depolarisation and Calcium Ion
Release

An action potential from a motor

neuron triggers the release of
acetylcholine into the motor end
plate

Acetylcholine initiates depolarisation

within the sarcolemma, which is
spread through the muscle fibre via
T tubules

Depolarisation causes the

sarcoplasmic reticulum to release
stores of calcium ions (Ca2+)
2. Actin and Myosin
Cross-Bridge Formation

On actin, the binding sites for

the myosin heads are covered
by a blocking complex
(troponin and tropomyosin)

Calcium ions bind to troponin

and reconfigure the complex,
exposing the binding sites for
the myosin heads

The myosin heads then form

a cross-bridge with the actin
filaments
3. Sliding Mechanism of Actin and
Myosin

ATP binds to the myosin head, breaking

the cross-bridge between actin and
myosin

ATP hydrolysis causes the myosin heads

to change position and swivel, moving
them towards the next actin binding site

The myosin heads bind to the new actin

sites and return to their original
conformation

This reorientation drags the actin along

the myosin in a sliding mechanism
4. Sarcomere Shortening

The repeated reorientation

of the myosin heads drags
the actin filaments along the
length of the myosin

As actin filaments are

anchored to Z lines, the
dragging of actin pulls the Z
lines closer together,
shortening the sarcomere

As the individual sarcomeres

become shorter in length, the
muscle fibres as a whole
contracts
Summery of skeletal muscle
contraction
What is the sliding filament
theory ?
 Skeletal muscle tissue Engineering
Skeletal muscle tissue, which comprises about 45% of total body
mass, is necessary for generating forces for movement and
locomotion
While skeletal muscle has an inherent ability to regenerate in
response to minor injuries

More severe conditions, such as myopathies, substantial

traumatic injury, aggressive tumor ablation, and prolonged
denervation, result in irreversible loss of muscle mass and
function

Deficiencies in regeneration often result in fibrous scar tissue

formation and fatty degeneration of muscle
 Tissue Engineering of Skeletal Muscle
In 1932, Sir Winston Churchill predicted that it will be possible in
future to grow and engineer muscle tissue in vitro. As a
confirmation of his prediction, Vandenburgh et al. observed
contracting muscle tissue engineered in vitro for the first time in
1988 (Vandenburgh, H.H. et al. 1988).
Only one year later – in 1989 – the group showed that mechanical
stimulation of embrionic myoblasts in vitro facilitates longitudinal
growth of engineered skeletal muscle tissue (Vandenburgh, H.H.
& Karlisch 1989).
This rapid development raised high expectations for future
clinical applications of tissue engineering (TE) of skeletal muscle.
Indeed, engineered muscle tissue could be used in a wide range of
clinical situations.
 Two general approaches of skeletal muscle tissue engineering.
(A) Muscle cells and other supporting cells are combined with biomaterials in vitro,
followed by transplantation either after extended culture to promote muscle formation,
or immediately.
(B) Biomaterials, either alone or combined with cytokines, growth factors, or cells
secreting paracrine signals, are delivered to the body to induce regeneration by host muscle
cells
Mechanical Properties
Common biomedical materials used in musculoskeletal tissue engineering,
including natural and synthetic materials
Collagen-based scaffolds in musculoskeletal tissue engineering
Two-dimensional (2D) nanomaterials are receiving
ever-increasing interests in biomedical applications owing to
their special physicochemical characteristics.

The ability to use this kind of 2D nanomaterials as drug

delivery systems, tumor photothermal/photodynamic therapy
agents, and diagnostic imaging agents give a strong incentive
to research their in vitro and in vivo toxicities.
Skeletal muscle precursor cells (myoblasts) in vitro

Immunofluorescent staining for desmin (green) an intermediate filament specifically

expressed in myogenic differentiation prior to the formation of myotubes and fusion
into muscle fibers. Nuclei are stained non-specifically with DAPI
(Diamidine-phenylindole-dihydrochloride; blue).
 3D skeletal Muscle
Generating human skeletal muscle models is instrumental
for investigating muscle pathology and therapy.

3D skeletal myogenic differentiation of pluripotent cells was

induced within hydrogels under tension to provide myofiber
alignment.

Artificial muscles recapitulated characteristics of human

skeletal muscle tissue and could be implanted into
immunodeficient mice.
Pathological cellular hallmarks of incurable forms of severe
muscular dystrophy could be modeled with high fidelity
using this 3D platform.
Schematic cartoon showing the cellular and engineered hydrogel strategy used to
obtain human myobundles
 Hydrogels
Recent developments in 3D cell culture hydrogel. ...
VitroGel™

Hydrogels are animal origin-free hydrogel systems closely

mimicking the natural extracellular matrix environment.

Cells grown in this 3D hydrogel possess morphologies

and cellular behaviors close to their natural state making it
ideal for 3D cell culture
Types of Hydrogel
Different types of hydrogels applicable for
tissue engineering.
Different designs for scaffolds currently used in
muscle TE
Concept of in vitro TE of skeletal muscle with three different
types of scaffolds
Summary
 The main challenges in skeletal muscle TE are therefore:
(1) engineering a suitable matrix for muscle TE including a
clinical application,

(2) Improving further myogenic differentiation in vivo and

(3) enabling the transplantation of functional skeletal muscle

tissue to the recipient site including microsurgical
anastomosis of an adequate vasculature as well as motoric
neurotization of the engineered muscle tissue.

Despite these obstacles, the achievements of the recent years

demonstrate an encouraging progress of skeletal muscle
TE research.
Cardiovascular Tissue Engineering
Biomaterials
Biomaterials- Scaffold
Cells
Biomolecules
Ophthalmic Tissue Engineering