Tissue Engineering and

Regenerative Medicine
A Biomedical and Classroom
Revolution
 Tissue Engineering and
Regenerative Medicine
1. It’s HOT!
2. It’s Relevant!
Everybody is a potential candidate for its
application.
It helps answer the dreaded question:
“Why do we have to learn all this stuff?”
It’s multidisciplinary, a new trend in science and
education
3. It’s a ‘Burgh Thing!
Five hottest jobs for the
next millennium will be
bioengineering/biomedical
related.

Tissue
Engineering
Hottest job for 21st
Century
What is Tissue Engineering?

 Broadly Defined: Tissue Engineering is

the development and manipulation of
artificial implants, laboratory-grown
tissues, genetically engineered cells
and/or molecules to replace or support the
function of defective or injured parts of
the body.
How we have define regenerative medicine?

Tissue Engineering and

Biomaterials

Cellular Therapies Medical Devices and Artificial

Organs
What is Tissue Engineering/
Regenerative Medicine?
Replacing diseased or injured
tissues with tissue constructs
designed and fabricated for the
specific needs of each individual
patient.
What are
Biomaterials?
Material intended to interface
with biological systems to
evaluate, treat, augment or
replace any tissue, organ or
function in the body.
 No One Discipline Can Tackle
the Problem Alone

Lee Weiss, Carnegie Mellon

Answering these questions requires the marriage of disciplines

Materials Molecular Cell

Science Biology Biology

Chemical Clinicians Biochemistry

Engineering

Robotics Computational
Genomics Biology
 Guided Tissue Repair

If needed, harvest cells

from patient.

Biomimetic Culture Implant

extracellular
matrix

Lee Weiss, Carnegie Mellon

 Variations
On a
Theme

Lee Weiss, Carnegie Mellon

 Principles of Tissue
Engineering

Cells ECM

Defect Regeneration

Hormones Blood
Supply

Phil Campbell, Carnegie Mellon

 Tissue Structure and Function
may be Compromised By:
1. Inherent design flaws
2. Hereditary/congenital defects or
conditions
3. Disease
4. Trauma
5. Environmental influences/insults
6. Aging
 Potential Solutions:
1. Surgical or physical manipulation
2. Drug therapy
3. Diet/lifestyle changes
4. Transplants
5. Artificial tissues/organs
6. Gene therapy
7. Tissue Engineering/Regenerative
Medicine
Forecasts of the American Population Aged 85 Years and Over
1960 -1990 Actual
2000, 2040 Census Bureau middle mortality
2040 Census Bureau low mortality
60
2040 Guralnik et al. 1998

50 2040 Manon et al. 1997

Population (million)

40

30

20

10

0
2040
60

70

80

90

00
19

19

19

19

20

Oxford Textbook of Geriatric Medicine 2000

 Medical costs

(1996 US dollars per capita)

USA $3898
% of gross domestic product in1996

16
United Kingdom $1317
14 Turkey $232
Health Expenditures

12
US Medicare expenditures for
10 last year of life doubles ages
8 65 - 69 years compared to 90+
years. (excluding nursing home
6 costs)
4
1987-1995 Hip replacements
2 among women rose from
0 143/100,000 to 1444/100,000
USA Germany Turkey
Switzerland Mexico
Canada South Korea
Poland
Oxford Textbook of Geriatric Medicine 2000
FDA approved products
Infuse Bone Graft
Bone morphogenetic protein-7,
Osteogenic peptide-1
Regranex
Carticel
Transcyte
Intergra Dermal Regeneration Template
Dermagraft
Apligraft
Ortec
 Apligraf is a living, bi-
layered skin substitute
consisting of living cells
and structural proteins.

Unlike human skin, Apligraf does not contain melanocytes,macrophages,

and lymphocytes, or other structures such as blood vessels, hair follicles
or sweat glands.
The ‘burgh, THEN….
Same area,
NOW…
Dr. Amit Patel: Cell Therapy for
Heart Failure
 Stephen Badylak, PhD,
MD, DVM
SIS ECM

 SIS, ECM for repair of soft tissues.

Once in place, the matrix, a 3-
dimensional scaffold void of cells
but with structural and functional
proteins still intact, serves to
recruit the appropriate cells for
tissue remodeling without
producing scarring.
First marine mammal
application of ECM tissue
repair!
Meet Liko, 3-year old dolphin at
Dolphin Quest on Hawaii’s Big
Island,
Liko sustained a tear at base of
his dorsal (top) fin -- likely in a
game of “chase” with his
dolphin cohorts.
Thanks to Dr. Badylak’s SIS
ECM, Liko has healed and is
again performing.
300,000
Patients
…
>5 Companies
>15 FDA allowances
Using Embryonic Stem Cells for TERM

Stem Cells: The

Key to Tissue
Design

•Cellular Biology
•Ethical
Implications
•Tissue Structure
& Function
Adult Stem Cells

Examples:

- Bone marrow –
derived
- Adipose-derived
- Muscle-derived
 An Ultimate Vision for Regenerative
Medicine: Complete Tissue Regeneration
Spinal Cord

Upper and
Tail Retina and Lens Lower Jaw
Heart

Limb

The Newt
From Dr. Susan Bryant,
AdaptedUniv.from
of Calif.,
BrockesIrvine
Phil Campbell, Carnegie Mellon
 Tissue Engineering Roadblocks
 Inadequate understanding of basic biology of
regenerative processes
 Lack of adequate biomimetic materials to act as
scaffolds for induction of regeneration in vivo, or to
build bioartificial tissues in vitro

 Inadequate cell sources for transplantation or building

bioartificial tissues
 Problem of immunosuppressive regimens introduced
by allogeneic and xenogeneic cells.

 Bioethical issues associated with the use of fetal and

embryonic stem cells as sources
 • The most critical roadblock to overcome
remains our inadequate understanding of the
basic biology…

Phil Campbell, Carnegie Mellon

TE in the Classroom: Approaches
 TE as Overall Theme in Biology
 Pick and Choose
 TE as reinforcer
 2+2+2 example
 Ready made unit
 TE Manual Overview
Tissue Engineering: Introduction
Tissue Structure and Function
 Tissue Origins
 Tissues in the Mature Body
 Tissue Development and Maintenance
 Stem Cells: The Keys to Tissue Design

Bone Tissue Engineering

 Bone Mechanics
 Porosity, Pore Size, and Surface Area
 Bone Composition
 Diffusion
 Cell Migration
 Cell Proliferation and Differentiation
Bone TE (cont) Student Activities:
 Activity 1: Build a Tissue
 Activity 2: Bone Strength
 Activity 3: Scaffold Diffusion Assay
 Activity 4: Biochemical Assay
 Activity 5: Cell Survival Assay
 Activity 6: Scaffold Synthesis and Characterization
 Activity 7: The Precarious Balance
Immunology and TE
 The Immune System
 Current Laboratory Techniques in Immunology
 Systemic Lupus Erythematosus
Student Activities
 Activity 1: Cells of the Immune System
 Activity 2: Immunohistology
 Activity 3: Complement
 Activity 4: The Chemotactic Response
 Activity 5: Immunogenetics of A.I.D.
Muscle Tissue Engineering
 Cell Culturing
 Muscle/Stem Cell Cultures
 Biochemical Identification/Characterization
 Therapeutic Disease Models
 Animal Model Therapy Assessment
Student Activities
 Activity 1: Chicken Little
 Activity 2: Muscle Repair
 Activity 3: Cell Culture and Differentiation
 Activity 4: Stem Cell Potential
 Activity 5: Stem Cell Seeding

 Assessment
 Glossary
 Supplementals, i.e. bioethics, activity extensions
 Standards
 Standards-Based: Examples
Chapter 1: Tissue Engineering: An Introduction
PA Standards Met: Refer to 3.8 Science, Technology, and Human
Endeavors (3.8.10 A, B, and C)
NSES Standards: Refer to E. Science and Technology; F. Science in
Personal and Social Perspectives

Chapter 2: Tissue Structure and Function

PA Standards Met: Refer to 3.1 Unifying Themes (3.1.10 A, B, C and
E) and
3.3 Biological Sciences (3.3.10 A and B)
NSES Standards: Refer to C. Life Science

Chapter 3: Overview of Classroom Activities

PA Standards Met: Refer to 3.2 Inquiry and Design (3.2.10 A, B, C and
D. These standards are the basis of all classroom demonstrations
and activities)
NSES Standards: Refer to A. Science as Inquiry
Load/Mass Ratio.
Provides insight
regarding
mechanical and
biological needs for
implanted scaffolds.
TE Triangle: Cells + Signals + Scaffold
How do growth factors interact with a scaffold? How does
combination of selected growth factors + scaffold affect stem
cell populations?

How are variables related? What are 3 common components

used to regenerate implantable tissue? What role do signals
play in the formation of functional tissue? What does the
standard curve allow us to quantify?

Objectives:
1. Create a standard curve that illustrates the relationship
between 2 variables.
2. Demonstrate the use and efficiency of a scaffold model.
3. Explain importance and function of cellular signals (growth
factors).
4. Students will understand the functional relationship of all of
the tissue engineering components (cells, signals, scaffolds)
Figure 1: Dilution series of Figure 2: Scaffold seeding growth
simulated growth factor solution factor by diffusion

Figure 3: Preparing scaffold growth Figure 4: Quantifying growth factor

factor leachettes for analysis scaffold seeding by spectrophotometery
 TE Triangle:
Procedure
Absorption Spectrum
1. Turn on spec and obtain sample of food coloring (label 100% concentration)
2. Transfer approximately 5 mL of this sample into a spec tube.
3. Set wavelength to 400 nm. Blank the machine with a tube of water.
4. Measure absorbance of your sample at this wavelength.
5. Set wavelength of machine to 420 nm. Blank as before and record absorbance.
6.Repeat at intervals of 20 nm up to 600 nm.
7.Graph results (this can be done later, but remember the absorbance maximum.)
The x-axis represents wavelength, and the y-axis represents absorbance.

Standard Curve Analysis

1. Create a series of dilutions of your original sample as directed by your teacher.
Be sure to label final concentration of each tube.
2. Set machine to absorbance maximum as determined in part A.
3. Measure absorbance of each dilution.
4. Graph data. X-axis represents concentration of your samples (dilutions), and Y-
axis represents absorbance. This is now your standard curve.
5.Obtain an ‘unknown’ sample of tissue extract from your teacher.
6.Measure the absorbance.
7.Using the standard curve, determine the concentration of biochemical ‘x’
 Immunology Classroom Activities
PCR Technology used to investigate genes
with possible correlations to SLE. PCR
profiles form family members afflicted with
SLE are generated and used as a means
of establishing correlation between the
gene and the presence of disease.
 Additional Resources/Activities
 Teacher Summer Institute, June 26-30, 2006,
 Middle School Summer Camp and Camp-on-Disc
 Planetarium Show w/DVD modules and website
 Virtual stem cell lab, Children’s Boston Hospital
www.childrenshospital.org/research/Site2029/m
ainpageS202P23sublevel39.html
Contact: PTEI, 100 Technology Drive, Pittsburgh,
PA 15219, 412/235-5230; www.ptei.org

NSTA BOOTH #2356