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Tissue Engineering and Regenerative Medicine

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Title: Tissue Engineering and Regenerative Medicine


1
Tissue Engineering and Regenerative Medicine
  • A Biomedical and Classroom
  • Revolution

2
Tissue Engineering and Regenerative Medicine
  • 1. Its HOT!
  • 2. Its Relevant!
  • Everybody is a potential candidate for its
    application.
  • It helps answer the dreaded question
  • Why do we have to learn all this stuff?
  • Its multidisciplinary, a new trend in science
    and education
  • 3. Its a Burgh Thing!

3
Five hottest jobs for the next millennium will
be bioengineering/biomedical related.
Tissue Engineering Hottest job for 21st Century
4
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5
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.

6
How we have define regenerative medicine?
Tissue Engineering and Biomaterials
7
What is Tissue Engineering/ Regenerative Medicine?
What are Biomaterials?
8
No One Discipline Can Tackle the Problem Alone
Lee Weiss, Carnegie Mellon
9
Answering these questions requires the marriage
of disciplines
Molecular Biology
Materials Science
Cell Biology
Clinicians
Biochemistry
Chemical Engineering
Computational Biology
Robotics
Genomics
10
Guided Tissue Repair
If needed, harvest cells from patient.
Growth factors
Cells
Biomimetic extracellular matrix
Implant
Culture
Lee Weiss, Carnegie Mellon
11
Variations On a Theme
Lee Weiss, Carnegie Mellon
12
Principles of Tissue Engineering
  • Cells

ECM
Defect
Regeneration
Blood Supply
Hormones
Phil Campbell, Carnegie Mellon
13
Tissue Structure and Function may be Compromised
By
  • Inherent design flaws
  • Hereditary/congenital defects or conditions
  • Disease
  • Trauma
  • Environmental influences/insults
  • Aging

14
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

15
Forecasts of the American Population Aged 85
Years and Over
Oxford Textbook of Geriatric Medicine 2000
16
Medical costs
(1996 US dollars per capita)
USA 3898 United Kingdom 1317 Turkey
232
US Medicare expenditures for last year of life
doubles ages 65 - 69 years compared to 90
years. (excluding nursing home costs)
1987-1995 Hip replacements among women rose from
143/100,000 to 1444/100,000
Oxford Textbook of Geriatric Medicine 2000
17
FDA approved products Infuse Bone Graft Bone
morphogenetic protein-7, Osteogenic
peptide-1 Regranex Carticel Transcyte Intergra
Dermal Regeneration Template Dermagraft Apligraft
Ortec
18
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.
19
The burgh, THEN.
20
Same area, NOW
21
Dr. Amit Patel Cell Therapy for Heart Failure
22
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.

23
(No Transcript)
24
First marine mammal application of ECM tissue
repair! Meet Liko, 3-year old dolphin at Dolphin
Quest on Hawaiis 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. Badylaks SIS ECM, Liko has healed
and is again performing.
25
300,000 Patients
gt5 Companies gt15 FDA allowances
26
Using Embryonic Stem Cells for TERM
  • Stem Cells The Key to Tissue Design
  • Cellular Biology
  • Ethical Implications
  • Tissue Structure Function

27
Adult Stem Cells
Examples - Bone marrow derived -
Adipose-derived - Muscle-derived
28
An Ultimate Vision for Regenerative Medicine
Complete Tissue Regeneration
Spinal Cord
Upper and Lower Jaw
Retina and Lens
Tail
Heart
Limb
The Newt
Adapted from Brockes
From Dr. Susan Bryant, Univ. of Calif., Irvine
Phil Campbell, Carnegie Mellon
29
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
30
  • The most critical roadblock to overcome remains
    our inadequate understanding of the basic
    biology

Phil Campbell, Carnegie Mellon
31
TE in the Classroom Approaches
  • TE as Overall Theme in Biology
  • Pick and Choose
  • TE as reinforcer
  • 222 example
  • Ready made unit

32
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

33
  • 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.

34
  • 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

35
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

36
Load/Mass Ratio. Provides insight regarding
mechanical and biological needs for implanted
scaffolds.
37
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)

38
Figure 1 Dilution series of simulated growth
factor solution
Figure 2 Scaffold seeding growth factor by
diffusion
Figure 4 Quantifying growth factor scaffold
seeding by spectrophotometery
Figure 3 Preparing scaffold growth factor
leachettes for analysis
39
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

40
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.

41
TODAY
TOMORROW
42
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, Childrens Boston Hospital
    www.childrenshospital.org/research/Site2029/mainpa
    geS202P23sublevel39.html
  • Contact PTEI, 100 Technology Drive, Pittsburgh,
    PA 15219, 412/235-5230 www.ptei.org
  • NSTA BOOTH 2356
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