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Tissue Engineering and Regenerative Medicine

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:

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Tissue Engineering and Regenerative Medicine

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

  4. 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.

  5. Medical Devices and Artificial Organs Cellular Therapies How we have define regenerative medicine? Tissue Engineering and Biomaterials

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

  7. No One Discipline Can Tackle the Problem Alone Lee Weiss, Carnegie Mellon

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

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

  10. Variations On a Theme Lee Weiss, Carnegie Mellon

  11. Principles of Tissue Engineering Cells ECM Defect Regeneration Blood Supply Hormones PhilCampbell, Carnegie Mellon

  12. Tissue Structure and Function may be Compromised By: • Inherent design flaws • Hereditary/congenital defects or conditions • Disease • Trauma • Environmental influences/insults • Aging

  13. Potential Solutions: • Surgical or physical manipulation • Drug therapy • Diet/lifestyle changes • Transplants • Artificial tissues/organs • Gene therapy • Tissue Engineering/Regenerative Medicine

  14. Forecasts of the American Population Aged 85 Years and Over Oxford Textbook of Geriatric Medicine 2000

  15. 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

  16. FDA approved products Infuse Bone Graft Bone morphogenetic protein-7, Osteogenic peptide-1 Regranex Carticel Transcyte Intergra Dermal Regeneration Template Dermagraft Apligraft Ortec

  17. 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.

  18. The ‘burgh, THEN….

  19. Same area, NOW…

  20. Dr. Amit Patel: Cell Therapy for Heart Failure

  21. 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.

  22. 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.

  23. 300,000 Patients… >5 Companies >15 FDA allowances

  24. Using Embryonic Stem Cells for TERM • Stem Cells: The Key to Tissue Design • Cellular Biology • Ethical Implications • Tissue Structure & Function

  25. Adult Stem Cells Examples: - Bone marrow – derived - Adipose-derived - Muscle-derived

  26. 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

  27. 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

  28. The most critical roadblock to overcome remains our inadequate understanding of the basic biology… PhilCampbell, Carnegie Mellon

  29. TE in the Classroom: Approaches • TE as Overall Theme in Biology • Pick and Choose • TE as reinforcer • 2+2+2 example • Ready made unit

  30. 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

  31. 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.

  32. 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

  33. 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

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

  35. 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)

  36. 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

  37. 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’

  38. 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.

  39. TODAY TOMORROW

  40. 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/mainpageS202P23sublevel39.html Contact: PTEI, 100 Technology Drive, Pittsburgh, PA 15219, 412/235-5230; www.ptei.org NSTA BOOTH #2356

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