Tissue Engineering Tissue Engineering and Regenerative and Regenerative

MedicineMedicine

A Biomedical and A Biomedical and Classroom Classroom

RevolutionRevolution

Tissue Engineering Tissue Engineering and Regenerative and Regenerative

MedicineMedicine1.1. It’s HOT!It’s HOT!2.2. It’s Relevant!It’s Relevant!

Everybody is a potential candidate for its Everybody is a potential candidate for its application.application.It helps answer the dreaded question:It helps answer the dreaded question:

““Why do we have to learn all this Why do we have to learn all this stuff?”stuff?”It’s multidisciplinary, a new trend in science It’s multidisciplinary, a new trend in science and educationand education

3. 3. It’s a ‘Burgh Thing!It’s a ‘Burgh Thing!

Tissue Engineering

Hottest job for 21st Century

Five hottest jobs for the next millennium will bebioengineering/biomedicalrelated.

What is Tissue Engineering?

Broadly Defined: Tissue Engineering Broadly Defined: Tissue Engineering is the development and manipulation is the development and manipulation of artificial implants, laboratory-grown of artificial implants, laboratory-grown tissues, genetically engineered cells tissues, genetically engineered cells and/or molecules to replace or support and/or molecules to replace or support the function of defective or injured the function of defective or injured parts of the body.parts of the body.

Cellular Therapies Medical Devices and Artificial Organs

How we have define regenerative medicine?

Tissue Engineering and Biomaterials

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.

What is Tissue Engineering/Regenerative Medicine?

No One Discipline Can Tackle the Problem Alone

Lee Weiss, Carnegie Mellon

ChemicalEngineering

Molecular Biology

GenomicsRobotics Computational

Biology

MaterialsScience

CellBiology

Clinicians Biochemistry

Answering these questions requires the marriage of disciplines

CellsGrowth factors

Biomimeticextracellular

matrix

Culture Implant

If needed, harvest cells from patient.

Guided Tissue RepairGuided Tissue Repair

Lee Weiss, Carnegie Mellon

VariationsOn a

Theme

Lee Weiss, Carnegie Mellon

CellsCells

Principles of Tissue Principles of Tissue EngineeringEngineering

ECMECM

HormoneHormoness

BloodBloodSupplySupply

Defect Regeneration

Phil Campbell, Carnegie Mellon

Tissue Structure and Tissue Structure and Function may be Function may be Compromised By:Compromised By:

1.1. Inherent design flawsInherent design flaws

2.2. Hereditary/congenital defects or Hereditary/congenital defects or conditionsconditions

3.3. DiseaseDisease

4.4. Trauma Trauma

5.5. Environmental influences/insultsEnvironmental influences/insults

6.6. AgingAging

Potential Solutions:Potential Solutions:

1.1. Surgical or physical manipulationSurgical or physical manipulation2.2. Drug therapyDrug therapy3.3. Diet/lifestyle changesDiet/lifestyle changes4.4. TransplantsTransplants5.5. Artificial tissues/organsArtificial tissues/organs6.6. Gene therapyGene therapy7.7. Tissue Engineering/Regenerative Tissue Engineering/Regenerative

MedicineMedicine

Forecasts of the American Population Aged 85 Years and Over

Oxford Textbook of Geriatric Medicine 2000

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USA $3898United Kingdom $1317Turkey $232

(1996 US dollars per capita)

1987-1995 Hip replacements among women rose from 143/100,000 to 1444/100,000

US Medicare expenditures for last year of life doubles ages 65 - 69 years compared to 90+ years. (excluding nursing home costs)

Medical costsMedical costs

Oxford Textbook of Geriatric Medicine 2000

FDA approved products

Infuse Bone GraftBone morphogenetic protein-7,

Osteogenic peptide-1RegranexCarticelTranscyteIntergra Dermal Regeneration TemplateDermagraftApligraftOrtec

Unlike human skin, Apligraf does not contain melanocytes,macrophages, and lymphocytes, or other structures such as blood vessels, hair follicles or sweat glands.

Apligraf is a living, bi-layered skin substitute consisting of living cells and structural proteins.

The ‘burgh, THEN….

Same area, NOW…

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

SIS, ECM for repair of soft SIS, ECM for repair of soft tissues. Once in place, the tissues. Once in place, the matrix, a 3-dimensional matrix, a 3-dimensional scaffold void of cells but with scaffold void of cells but with structural and functional structural and functional proteins still intact, serves to proteins still intact, serves to recruit the appropriate cells for recruit the appropriate cells for tissue remodeling without tissue remodeling without producing scarring. producing scarring.

Stephen Badylak, PhD, MD, DVM

SIS ECM

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.

300,000 300,000 PatientsPatients

……>5 Companies>15 FDA allowances

Stem Cells: The Key to Tissue Design

•Cellular Biology

•Ethical Implications

•Tissue Structure & Function

Using Embryonic Stem Cells for TERM

Adult Stem Cells

Examples:

- Bone marrow – derived - Adipose-derived- Muscle-derived

Spinal CordSpinal CordSpinal CordSpinal Cord

Upper and Upper and Lower JawLower JawUpper and Upper and Lower JawLower Jaw

LimbLimbLimbLimb

Retina and LensRetina and LensRetina and LensRetina and Lens

An Ultimate Vision for Regenerative Medicine: Complete Tissue Regeneration

An Ultimate Vision for Regenerative Medicine: Complete Tissue Regeneration

Adapted from BrockesAdapted from Brockes

TailTail TailTail HeartHeartHeartHeart

The NewtThe NewtFrom Dr. Susan Bryant, Univ. of Calif., Irvine

Phil Campbell, Carnegie MellonPhil Campbell, Carnegie Mellon

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

Tissue Engineering Roadblocks

• The most critical roadblock to overcome remains our inadequate understanding of the basic biology…

Phil Campbell, Carnegie Mellon

TE in the Classroom: TE in the Classroom: ApproachesApproaches

TE as Overall Theme in TE as Overall Theme in BiologyBiology

Pick and ChoosePick and Choose TE as reinforcerTE as reinforcer 2+2+2 example2+2+2 example Ready made unitReady made unit

TE Manual OverviewTE Manual OverviewTissue Engineering: IntroductionTissue Engineering: IntroductionTissue Structure and FunctionTissue Structure and Function Tissue Origins Tissue Origins Tissues in the Mature BodyTissues in the Mature Body Tissue Development and MaintenanceTissue Development and Maintenance Stem Cells: The Keys to Tissue DesignStem Cells: The Keys to Tissue Design

Bone Tissue EngineeringBone Tissue Engineering Bone Mechanics Bone Mechanics Porosity, Pore Size, and Surface Area Porosity, Pore Size, and Surface Area Bone Composition Bone Composition Diffusion Diffusion Cell Migration Cell Migration Cell Proliferation and DifferentiationCell Proliferation and Differentiation

Bone TE (cont) Bone TE (cont) Student Activities:Student Activities: Activity 1: Build a Tissue Activity 1: Build a Tissue Activity 2: Bone StrengthActivity 2: Bone Strength Activity 3: Scaffold Diffusion AssayActivity 3: Scaffold Diffusion Assay Activity 4: Biochemical AssayActivity 4: Biochemical Assay Activity 5: Cell Survival AssayActivity 5: Cell Survival Assay Activity 6: Scaffold Synthesis and CharacterizationActivity 6: Scaffold Synthesis and Characterization Activity 7: The Precarious BalanceActivity 7: The Precarious BalanceImmunology and TEImmunology and TE The Immune SystemThe Immune System Current Laboratory Techniques in ImmunologyCurrent Laboratory Techniques in Immunology Systemic Lupus ErythematosusSystemic Lupus ErythematosusStudent ActivitiesStudent Activities Activity 1: Cells of the Immune System Activity 1: Cells of the Immune System Activity 2: Immunohistology Activity 2: Immunohistology Activity 3: Complement Activity 3: Complement Activity 4: The Chemotactic ResponseActivity 4: The Chemotactic Response Activity 5: Immunogenetics of A.I.D.Activity 5: Immunogenetics of A.I.D.

Muscle Tissue EngineeringMuscle Tissue Engineering Cell CulturingCell Culturing Muscle/Stem Cell CulturesMuscle/Stem Cell Cultures Biochemical Identification/CharacterizationBiochemical Identification/Characterization Therapeutic Disease ModelsTherapeutic Disease Models Animal Model Therapy AssessmentAnimal Model Therapy AssessmentStudent ActivitiesStudent Activities Activity 1: Chicken LittleActivity 1: Chicken Little Activity 2: Muscle RepairActivity 2: Muscle Repair Activity 3: Cell Culture and DifferentiationActivity 3: Cell Culture and Differentiation Activity 4: Stem Cell PotentialActivity 4: Stem Cell Potential Activity 5: Stem Cell SeedingActivity 5: Stem Cell Seeding

AssessmentAssessment GlossaryGlossary Supplementals, i.e. bioethics, activity extensionsSupplementals, i.e. bioethics, activity extensions StandardsStandards

Standards-Based: ExamplesStandards-Based: ExamplesChapter 1: Tissue Engineering: An IntroductionChapter 1: Tissue Engineering: An IntroductionPA Standards MePA Standards Met: Refer to 3.8 Science, Technology, and t: Refer to 3.8 Science, Technology, and

Human Endeavors (3.8.10 A, B, and C)Human Endeavors (3.8.10 A, B, and C)NSES StandardsNSES Standards: Refer to E. Science and Technology; F. : Refer to E. Science and Technology; F.

Science in Personal and Social PerspectivesScience in Personal and Social Perspectives

Chapter 2: Tissue Structure and FunctionChapter 2: Tissue Structure and FunctionPA Standards MetPA Standards Met: Refer to 3.1 Unifying Themes (3.1.10 A, B, : Refer to 3.1 Unifying Themes (3.1.10 A, B,

C and E) and C and E) and 3.3 Biological Sciences (3.3.10 A and B)3.3 Biological Sciences (3.3.10 A and B)NSES StandardsNSES Standards: Refer to C. Life Science: Refer to C. Life Science

Chapter 3: Overview of Classroom ActivitiesChapter 3: Overview of Classroom ActivitiesPA Standards MetPA Standards Met: Refer to 3.2 Inquiry and Design (3.2.10 A, : Refer to 3.2 Inquiry and Design (3.2.10 A,

B, C and D. These standards are the basis of all classroom B, C and D. These standards are the basis of all classroom demonstrations and activities)demonstrations and activities)

NSES StandardsNSES Standards: Refer to A. Science as Inquiry: Refer to A. Science as Inquiry

Load/Mass Ratio.

Provides insight regarding mechanical and biological needs for implanted scaffolds.

TE Triangle: Cells + Signals + TE Triangle: Cells + Signals + ScaffoldScaffold

How do growth factors interact with a scaffold? How does How do growth factors interact with a scaffold? How does combination of selected growth factors + scaffold affect combination of selected growth factors + scaffold affect stem cell populations? stem cell populations?

How are variables related? What are 3 common How are variables related? What are 3 common components used to regenerate implantable tissue? What components used to regenerate implantable tissue? What role do signals play in the formation of functional tissue? role do signals play in the formation of functional tissue? What does the standard curve allow us to quantify? What does the standard curve allow us to quantify?

Objectives:Objectives:1. Create a standard curve that illustrates the relationship 1. Create a standard curve that illustrates the relationship

between 2 variables. between 2 variables. 2. Demonstrate the use and efficiency of a scaffold model.2. Demonstrate the use and efficiency of a scaffold model.3. Explain importance and function of cellular signals 3. Explain importance and function of cellular signals

(growth factors).(growth factors).4. Students will understand the functional relationship of all 4. Students will understand the functional relationship of all

of the tissue engineering components (cells, signals, of the tissue engineering components (cells, signals, scaffolds)scaffolds)

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

TE Triangle:TE Triangle:ProcedureProcedureAbsorption Spectrum Absorption Spectrum 1. Turn on spec and obtain sample of food coloring (label 100% 1. Turn on spec and obtain sample of food coloring (label 100%

concentration)concentration)2. Transfer approximately 5 mL of this sample into a spec tube.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.3. Set wavelength to 400 nm. Blank the machine with a tube of water.4. Measure absorbance of your sample at this wavelength.4. Measure absorbance of your sample at this wavelength.5. Set wavelength of machine to 420 nm. Blank as before and record 5. Set wavelength of machine to 420 nm. Blank as before and record

absorbance.absorbance.6.Repeat at intervals of 20 nm up to 600 nm.6.Repeat at intervals of 20 nm up to 600 nm.7.Graph results (this can be done later, but remember the absorbance 7.Graph results (this can be done later, but remember the absorbance

maximum.) The x-axis represents wavelength, and the y-axis represents maximum.) The x-axis represents wavelength, and the y-axis represents absorbance.absorbance.

Standard Curve Analysis Standard Curve Analysis 1. Create a series of dilutions of your original sample as directed by your 1. Create a series of dilutions of your original sample as directed by your

teacher. Be sure to label final concentration of each tube. teacher. Be sure to label final concentration of each tube. 2. Set machine to absorbance maximum as determined in part A.2. Set machine to absorbance maximum as determined in part A.3. Measure absorbance of each dilution.3. Measure absorbance of each dilution.4. Graph data. X-axis represents concentration of your samples (dilutions), 4. Graph data. X-axis represents concentration of your samples (dilutions),

and Y-axis represents absorbance. This is now your standard curve.and Y-axis represents absorbance. This is now your standard curve.5.Obtain an ‘unknown’ sample of tissue extract from your teacher.5.Obtain an ‘unknown’ sample of tissue extract from your teacher.6.Measure the absorbance.6.Measure the absorbance.7.Using the standard curve, determine the concentration of biochemical ‘x’ 7.Using the standard curve, determine the concentration of biochemical ‘x’

Immunology Classroom Immunology Classroom ActivitiesActivities

PCR Technology used to investigate PCR Technology used to investigate genes with possible correlations to genes with possible correlations to SLE. PCR profiles form family SLE. PCR profiles form family members afflicted with SLE are members afflicted with SLE are generated and used as a means of generated and used as a means of establishing correlation between the establishing correlation between the gene and the presence of disease.gene and the presence of disease.

TODAY

TOM

ORRO

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Additional Additional Resources/ActivitiesResources/Activities Teacher Summer Institute, June 26-30, 2006, Teacher Summer Institute, June 26-30, 2006,

Middle School Summer Camp and Camp-on-DiscMiddle School Summer Camp and Camp-on-Disc Planetarium Show w/DVD modules and websitePlanetarium Show w/DVD modules and website Virtual stem cell lab, Children’s Boston Hospital Virtual stem cell lab, Children’s Boston Hospital

www.childrenshospital.org/research/Site2029/mawww.childrenshospital.org/research/Site2029/mainpageS202P23sublevel39.htmlinpageS202P23sublevel39.html

Contact:Contact: PTEI, 100 Technology Drive, Pittsburgh, PTEI, 100 Technology Drive, Pittsburgh, PA 15219, 412/235-5230; www.ptei.orgPA 15219, 412/235-5230; www.ptei.org

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