Tissue Engineering and Regenerative Medicine

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Tissue Engineering and Regenerative Medicine

• A Biomedical and Classroom
• Revolution

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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!

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

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

• Cells

ECM
Defect
Regeneration
Blood Supply
Hormones
Phil Campbell, Carnegie Mellon
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Tissue Structure and Function may be Compromised
By

• Inherent design flaws
• Hereditary/congenital defects or conditions
• Disease
• Trauma
• Environmental influences/insults
• Aging

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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

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Forecasts of the American Population Aged 85
Years and Over
Oxford Textbook of Geriatric Medicine 2000
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Medical costs
(1996 US dollars per capita)
USA 3898 United Kingdom 1317 Turkey
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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
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FDA approved products Infuse Bone Graft Bone
morphogenetic protein-7, Osteogenic
peptide-1 Regranex Carticel Transcyte Intergra
Dermal Regeneration Template Dermagraft Apligraft
Ortec
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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.
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The burgh, THEN.
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Same area, NOW
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Dr. Amit Patel Cell Therapy for Heart Failure
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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.

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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.
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300,000 Patients
gt5 Companies gt15 FDA allowances
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Using Embryonic Stem Cells for TERM

• Stem Cells The Key to Tissue Design
• Cellular Biology
• Ethical Implications
• Tissue Structure Function

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Adult Stem Cells
Examples - Bone marrow derived -
Adipose-derived - Muscle-derived
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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
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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
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• The most critical roadblock to overcome remains
  our inadequate understanding of the basic
  biology

Phil Campbell, Carnegie Mellon
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TE in the Classroom Approaches

• TE as Overall Theme in Biology
• Pick and Choose
• TE as reinforcer
• 222 example
• Ready made unit

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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

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• 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.

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• 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

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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

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

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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
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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

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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.

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TODAY
TOMORROW
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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