Upload
marlee-reader
View
217
Download
0
Tags:
Embed Size (px)
Citation preview
Declaration of Conflict of Interest or Relationship
Speaker Name: Richard G. SpencerSpeaker Name: Richard G. Spencer
I have no conflicts of interest to disclose with regard to the subject matter ofI have no conflicts of interest to disclose with regard to the subject matter ofthis presentation.this presentation.
Unmet Needs for MR in the UnsolvedUnmet Needs for MR in the UnsolvedProblem of Tissue Engineering:Problem of Tissue Engineering:
Richard G. SpencerRichard G. SpencerMagnetic Resonance Imaging and Spectroscopy SectionMagnetic Resonance Imaging and Spectroscopy Section
National Institute on Aging, NIHNational Institute on Aging, NIHBaltimore, MDBaltimore, MD
Not just one problem, not just one solutionNot just one problem, not just one solution
ISMRM Toronto, 2008ISMRM Toronto, 2008
Definition of Tissue Engineering:Definition of Tissue Engineering:
"…an interdisciplinary field that applies the"…an interdisciplinary field that applies theprinciples of engineering and life sciencesprinciples of engineering and life sciences
towards the development of biological substitutes that towards the development of biological substitutes that restore, maintain, or improve tissue function."restore, maintain, or improve tissue function."
Langer and Vacanti, 1993Langer and Vacanti, 1993
Includes:Includes: Tissue EngineeringTissue Engineering
Excludes: Excludes: Organ transplantsOrgan transplants Artificial mechanical organsArtificial mechanical organs Metal jointsMetal joints Hemodialysis, external blood oxygenators,…Hemodialysis, external blood oxygenators,…
Implanted cells which will develop function in vivoImplanted cells which will develop function in vivo--stem cells for MI or stroke repair --stem cells for MI or stroke repair --cartilage matrix from implanted cells--cartilage matrix from implanted cells
Implanted acellular scaffoldsImplanted acellular scaffolds--bioactive bone graft scaffold--bioactive bone graft scaffold
Implanted cellular constructs designed in the labImplanted cellular constructs designed in the lab----off-the-shelf cartilage, pancreas, liver,…off-the-shelf cartilage, pancreas, liver,…
Motivation for TEMotivation for TE
Limitations of non-biologic repair:Limitations of non-biologic repair:
Duration < longevity of patient Duration < longevity of patient (metal implants)(metal implants)
Highly invasive Highly invasive (hemodialysis)(hemodialysis)
Typically, no growth or adaptation Typically, no growth or adaptation (c.f. advantage of demand pacemakers)(c.f. advantage of demand pacemakers)
Funding OpportunitiesFunding Opportunities
NIH:NIH: regenerative medicine funding 2008-- regenerative medicine funding 2008-- US$575 M (approx. 2% of NIH budget)US$575 M (approx. 2% of NIH budget)
DoD:DoD: Armed Forces Institute of Regenerative Armed Forces Institute of Regenerative Medicine (AFIRM), April, 2008: Medicine (AFIRM), April, 2008: US$265 M over 5 yearsUS$265 M over 5 years
Other: Other: NIST, NSF, Arthritis Foundation, NIST, NSF, Arthritis Foundation, Musculoskeletal Transplant Foundation…Musculoskeletal Transplant Foundation…
Medtronic: INFUSE bone graftsMedtronic: INFUSE bone grafts
RESTORE small intestine submucosa (duPont)RESTORE small intestine submucosa (duPont)
Apligraf skin patch (Organogenesis)Apligraf skin patch (Organogenesis)
Cord blood stem cell bankingCord blood stem cell banking
The product list is limited but activeThe product list is limited but active
Lysaght, Tissue Engineering, 2008Lysaght, Tissue Engineering, 2008
(SIS)
# of firms or business units > 50# of firms or business units > 50
# of employees > 3000# of employees > 3000
Annual sales > US$1.3 billionAnnual sales > US$1.3 billion
# of patients treated > 1M# of patients treated > 1M
# products in preclinical stages > 50# products in preclinical stages > 50
Private sector activity, 2007 (worldwide)Private sector activity, 2007 (worldwide)
Lysaght, Tissue Engineering, 2008Lysaght, Tissue Engineering, 2008
"Structural imaging" may be less important "Structural imaging" may be less important than other capabilities of MR, such asthan other capabilities of MR, such as
more specific microstructural and molecular more specific microstructural and molecular analyses, and metabolic analysis analyses, and metabolic analysis
What Can MRI and MRS Contribute?What Can MRI and MRS Contribute?
Non-invasive Non-invasive Monitoring after Implantation Monitoring after Implantation Positional stabilityPositional stability Integration with surroundingsIntegration with surroundings Metabolic activityMetabolic activity Need for and response to ongoing interventionNeed for and response to ongoing intervention
Examples of Current MR/TE StudiesExamples of Current MR/TE Studies
TE Cartilage Development in Hollow-fiber BioreactorTE Cartilage Development in Hollow-fiber Bioreactor
● from bovine cartilage
● 150 million cells/mL● Cultured for 4 weeks ● Sectioned 5mm each
A B C D E F G H I J
OutflowInflow BR3
● from bovine cartilage
● 150 million cells/mL● Cultured for 4 weeks ● Sectioned 5mm each
A B C D E F G H I J
OutflowInflow BR3
Bovine neocartilageBovine neocartilagedeveloping from developing from
chondrocyteschondrocytes
ATP
PCr
MDP
TR (s)
2.5
Pi
3131P spectrum of avian neocartilageP spectrum of avian neocartilage
30
20
10
1 / T
2, s
-1
0.150.100.050.00Tissue GAG Concentration, w/w
R = 0.81, p < 0.0011 week 2 weeks
3 weeks 4 weeks
Blue: MT< 0.4 Red: MT> 0.8
MT of TE CartilageMT of TE Cartilage
P = 0.04
0.0
1.0
2.0
Control Ibuprofen
MT
C k
m, s
-1
Effect of ibuprofen on Effect of ibuprofen on developing cartilagedeveloping cartilage
Day 14Day 14Matrix containing Matrix containing iron-labeled cellsiron-labeled cells Non-labeled matrixNon-labeled matrix
Prussian blue Prussian blue stained sectionstained sectionMR ImageMR Image
FCDFCD S-GAG/S-GAG/Dry Wt.Dry Wt.
Dynamic Dynamic ModulusModulus
Equilibrium Equilibrium ModulusModulus
Collagen/Collagen/Dry Wt.Dry Wt.
Effect of chondroitinase on developing cartilageEffect of chondroitinase on developing cartilage
Neocartilate in BioreactorNeocartilate in Bioreactor
culturemedium
collagen I gel
5x5x1 mm voxel
time
internal chemical shift standard TSP 10mM
glucose diffusion front
Metabolic Mapping in TE cartilage Non MRI: Model Based Estimates
(Sengers B. G. et al. JBME 2005)
Fick’s law
Michaelis-Menten
Computed spatial glucose concentration
Reference geometryof collagen I gel
Time course of glucose diffusion into ROI_1
Direct measurement of spatial glucose concentration
1H Localized Spectroscopy in Collagen I Constructs
Courtesy David ReiterCourtesy David Reiter
100µm
defect regionrepair tissue
T2 = 49.5 ms
repair tissuedefect region
T2 = 83.4 ms
repair tissue
native cartilage
defect
Courtesy S. Ramaswamy, Bioengineering, Univ of Pittsburgh
Hydrogel defect repair in rabbitHistology vs MRI
Cartilage in hydrogelBiochem vs MRI
9 days 21 days 28 days 43 days
100µm
T2 = -1.0144 X (% tissue fill) + 121.12
R2 = 0.6715
10
20
30
40
50
60
70
80
90
100
30 40 50 60 70 80 90 100
% Tissue fill
Tra
nsv
erse
rel
ax
ati
on
tim
e,
T2
(ms)
10 mM CuCl2 10 mM CuCl2 landmark bulblandmark bulb
Defect 1Defect 1
Subchondral boneSubchondral bone
Defect 2Defect 2CartilageCartilage
TidemarkTidemark
Cartilage/Hydrogel StudiesCartilage/Hydrogel Studies
Asssessment of Cartilage formed inSelf-Aggregating Suspension Culture
MR-based methods offer advantages of providing noninvasive,
nondestructive, and quantitative surrogate measures of material
properties.
Characteristics of Culture Model Grows in suspension at high
density (1-2 X107 cells/ml) Forms a mass quickly (110 weeks mass >6 fold) Maintains cartilage phenotype(cartilage specific ECM, no Coll I )
Courtesy of George DodgeBone & Cartilage Research Lab, Nemours Biomedical Research
T1ρ = 105 ms T1ρ = 107 ms
Bovine patellar cartilage Engineered cartilage Previous studies: Direct correlation of T1ρ MR measurements with FCD and PG in cartilage
Source: Trattnig et al., 2007
MRI Following Matrix-assisted Autologous Chondrocyte TransplantationMRI Following Matrix-assisted Autologous Chondrocyte Transplantation
4 Weeks Post-Surgery:4 Weeks Post-Surgery:
Fluid-like contrast in repair zoneFluid-like contrast in repair zone
24 Weeks Post-Surgery:24 Weeks Post-Surgery:
Heterogeneous contrastHeterogeneous contrast
52 Weeks Post-Surgery:52 Weeks Post-Surgery:
Repair zone isointense Repair zone isointense with native cartilagewith native cartilage
TT22-weighted fast spin echo images reveal longitudinal changes in -weighted fast spin echo images reveal longitudinal changes in
repair zone containing implanted chondrocytesrepair zone containing implanted chondrocytes
TT11 weighted image weighted image
of of osteochondral plugosteochondral plug
Courtesy Richard Magin, UICCourtesy Richard Magin, UIC
TT11 and T and T22 values for values for
developing TE bonedeveloping TE bone
Conclusion: TConclusion: T22 is an is an
effective marker for effective marker for ossificationossification
11.7 T SE; TE/TR=1000/3011.7 T SE; TE/TR=1000/30
Ostogenesis in a Tissue Engineered ConstructOstogenesis in a Tissue Engineered Construct
MSC’s from adult bone marrow gelatin sponge osteogenic cx medium
constructfat
Courtesy of Hai-Ling Margaret Cheng, PhDThe Hospital for Sick Children and University of Toronto
Non-invasive MRI of tissue-engineered bladder construct
Major challenge to successful regeneration: creating immediate angiogenesisRole for imaging: non-invasive quantitative assessment of angiogenesis
Exposed bladder Angiogenesis at different VEGF levelsCD31-immunostaining
VEGF microvessel density
0
10
20
30
5 7 9 11 13
AU
C1m
in r = 0.784, P = 0.003
MVD (%)
r = 0.437, P = 0.1790
10
20
30
40
50
5 7 9 11 13
Gadomer
Gd-DTPA
MVD (%)
v p (
%)
r = 0.696, P = 0.012
r = 0.228, P = 0.50
Gadomer
Gd-DTPA
Tissue-engineered bladder construct
Gadomer (blood pool cx agent): correlation with microvessel density (MVD) through either fractional plasma volume (vp) or area under uptake curve (AUC); superior to Gd-DTPA
Gd-DTPA (extracellular cx agent): correlation with MVD is best obtained with AUC
SIGNIFICANCE: MRI measurement of absolute blood volume is feasible in therapeutic angiogenesis paradigms
Constantinidis et al. et al.
MR Monitors Viable Cell Number, Bioenergetics and Oxygen Distribution MR Monitors Viable Cell Number, Bioenergetics and Oxygen Distribution for Mouse Insulinoma for Mouse Insulinoma TC Cells in Tissue-Engineered PancreasTC Cells in Tissue-Engineered Pancreas
31P 1D CSI yields bioenergetic data with both spatial and temporal
resolution.
T1 of 19F PFTBA shows changes in dissolved oxygen content during hypoxic intervention.
Stem Cell Therapeutics—Cell PersistenceStem Cell Therapeutics—Cell Persistence
D. Kraitchman--JHUD. Kraitchman--JHU
Hypointense regions showing sites ofHypointense regions showing sites ofmesenchymal stem cell injectionmesenchymal stem cell injection
Loss of cells Loss of cells from from
noninfarctednoninfarctedtissuetissue
Tissue properties can often be evaluated Tissue properties can often be evaluated by highly by highly sensitivesensitive
but but invasiveinvasive techniques techniques
Gene expression analysisGene expression analysis for genotype and tissue quality for genotype and tissue quality
Electron microscopyElectron microscopy for microstructure for microstructure
HistologyHistology for exquisite structural detail for exquisite structural detail
ImmunohistochemistryImmunohistochemistry for protein expression for protein expression
Optical spectroscopyOptical spectroscopy for more specific matrix components for more specific matrix components
Microelectrodes / biochemical analysisMicroelectrodes / biochemical analysis for metabolic analysis for metabolic analysis
Mechanical testing Mechanical testing for functional support propertiesfor functional support properties
Unmet NeedsUnmet NeedsReplacement of gold-standard techniques with non-invasive versionsReplacement of gold-standard techniques with non-invasive versions
Molecule-specific MR:Molecule-specific MR: example: molecular imaging for collagen I vs collagen IIexample: molecular imaging for collagen I vs collagen II
Mechanical measurements: Mechanical measurements: example: bone/cartilage elastography; strain measurementsexample: bone/cartilage elastography; strain measurements
Sequences and analyses more specific to tissue componentsSequences and analyses more specific to tissue components example: water compartment analysis; multiparametric approachesexample: water compartment analysis; multiparametric approaches
Functional analysis (esp. metabolism)Functional analysis (esp. metabolism) example: localized spectroscopy for cell number and viabilityexample: localized spectroscopy for cell number and viability
Histologic-like resolutionHistologic-like resolution example: microscopyexample: microscopy
Opportunities Abound!Opportunities Abound!
Caveat:Caveat:
Capital value of publicly traded TE companies:Capital value of publicly traded TE companies:
2000: $2.5 billion US2000: $2.5 billion US
2003: $300 million US2003: $300 million US
2007: $4.7 billion US2007: $4.7 billion US
Lysaght, Tissue Engineering, 2008Lysaght, Tissue Engineering, 2008
Tissue engineering is currently in a growth phaseTissue engineering is currently in a growth phase
ConclusionsConclusions
MR can actually contribute to further progressMR can actually contribute to further progress
Fundable through several sourcesFundable through several sources
There is a substantial market presence There is a substantial market presence (contrast gene therapy)(contrast gene therapy)
Come to the educational session in Hawaii!Come to the educational session in Hawaii!