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

2. 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!

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

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
Medical Devices and Artificial Organs
Cellular Therapies

7. What is Tissue Engineering/ Regenerative Medicine?
Material intended to interface with biological systems to evaluate, treat, augment or replace any tissue, organ or function in the body.
Replacing diseased or injured tissues with tissue constructs designed and fabricated for the specific needs of each individual patient.
What are
Biomaterials?

8. No One Discipline Can Tackle
the Problem Alone
REPRESENTATIVES FROM EACH SCHOOL: CIT, MCS, AND SCS
FRPM EACH ENGINEERING DEPARTMENT
HERE IS WHERE TO SAY THAT PEPOLE LIKE ANDY MOORE MIGHT ALSO CONTRIBUTE; OR EVEN VLADIMER
THIS WAS A TEAM OF BREATH, BUT NOT DEPTH NOR ONE WITH THE BIOLOGY/CLINICAL BACKGROUND….NEEDED A LEARDER IN THIS FIELD TO DO IN A TIMELY FASHION
Lee Weiss, Carnegie Mellon

9. Answering these questions requires the marriage of disciplines
Materials
Science
Molecular Biology
Cell
Biology
Clinicians
Chemical
Engineering
Biochemistry
Robotics
Computational
Biology
Genomics

10. Guided Tissue Repair If needed, harvest cells from patient.
Growth factors
Cells
Biomimetic
extracellular
matrix
Culture
Implant
Lee Weiss, Carnegie Mellon

11. Lee Weiss, Carnegie Mellon
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: Surgical or physical manipulation Drug therapy
Diet/lifestyle changes
Transplants
Artificial tissues/organs
Gene therapy
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 years compared to 90+ years. (excluding nursing home costs)
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.

24. 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.

25. 300,000 Patients…
>5 Companies
>15 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
Tail
Retina and Lens
Heart
(VIDEO IS OVER 20 DAYS):
The ultimate vision is to break the genetic code to get complete regeneration….after all, Newt engineers have figured this out!
Okay, so this will not happen in my lifetime
Limb
The Newt
From Dr. Susan Bryant, Univ. of Calif., Irvine
Adapted from Brockes
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. Phil Campbell, Carnegie Mellon
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
2+2+2 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 ( 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 ( A, B, C and E) and
3.3 Biological Sciences ( 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 ( 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 3: Preparing scaffold growth factor leachettes for analysis
Figure 4: Quantifying growth factor scaffold seeding by spectrophotometery

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, Children’s Boston Hospital
Contact: PTEI, 100 Technology Drive, Pittsburgh, PA 15219, 412/ ;
NSTA BOOTH #2356