synthetic extracellular matrices for promoting angiogenesis eduardo a. silva harvard university,...
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SYNTHETIC EXTRACELLULAR MATRICES FOR PROMOTING ANGIOGENESIS
Eduardo A. SilvaEduardo A. Silva
Harvard University, Cambridge, MA, USAHarvard University, Cambridge, MA, USAandand
Faculdade de Engenharia da Universidade do Porto, Porto, PortugalFaculdade de Engenharia da Universidade do Porto, Porto, Portugal
December 2004December 2004Porto, PortugalPorto, Portugal
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MOTIVATION
Cardiovascular Diseases:
- Principal cause of mortality in western countries
Current Approaches to treat Ischemic Heart Diseases:
Pharmacologic
Invasive Surgical
Therapeutic Angiogenesis
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ISCHEMIA
Carmeliet & Jain Nature, 2000
Ischemia - isch- is restriction, hemia or haemia is blood
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ANGIOGENESISSprouting of new blood vessels from
pre-existent vessels
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VASCULAR ENDOTHELIAL GROWTH FACTOR
VEGF
Is a key regulator of blood vessels formation and function
Endothelial Cells are the primary target of VEGF
45 kDa homodimer
Four isoforms obtained by alternative splicing:
VEGF121, VEGF165, VEGF189, VEGF206
Muller et al., Structure, 5:1325-1338 (1997)Muller et al., Structure, 5:1325-1338 (1997) Fairbrother et al., Structure, 6:637-648 (1998)
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AIM I: Investigate VEGF signaling in cardiac cells under hypoxia.
AIM I
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VEGF SECRETION UNDER HYPOXIA
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TISSUE ENGINEERING STRATEGY
Localized, sustained delivery from bioactive polymeric systems
Constant local concentration
Protection from degradation
Minimal side effects
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AIM II
Development of an injectable polymeric system for controlled and defined delivery of angiogenic molecules
VEGF incorporated in an injectable polymer
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POLYMERIC SYSTEMAlginate
OOH
CO2-
O
O
OH
-O2C
OH
O
HO
CO2- OH
OHO
CO2-
OHO
O
OOH
OHO
CO2-
O
OH
G G MM G
Ca2+
-L- Guluronic acid
-D- Mannuronic acid
Gelation process
Hydrogel
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MODEL SYSTEM Distinct approach used to incorporate VEGF
Simple mixing VEGF and alginate
Pre-incorporation of VEGF in polymeric microspheres and immobilization in the alginate
+ +Ca2+
Angiogenic moleculesAlginate
+ + Microspheres
Polymeric system
Bioactive moleculesBioactive molecules
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ALGINATE MODIFICATIONSCombination of binary molecular weight distribution and partial oxidation
Oxidation with NaIOOxidation with NaIO44
Irradiation (5 Mrad)Irradiation (5 Mrad)
CHOCHO
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Solution Properties
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Gel Properties
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VEGF RELEASE
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BIOACTIVITY OF VEGF RELEASED
0
5000
10000
15000
20000
25000
0 1 4
Time (days)
Cell number
MediaMedia/VEGFMedia/VEGF released
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ANGIOGENESIS
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PDGF RELEASE
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VEGF/PDGF RELEASE
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In vivo evaluation of VEGF releasing materials
Model: Ischemic hind limb of Apo E null (Apo E -/-)
LDPI image
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In vivo evaluation of VEGF releasing materials
Blank
VEGF&PDGF
VEGF
Week 2 Week 4 Week 6
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In vivo evaluation of VEGF releasing materials
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CONCLUSIONS
Sustainable and controlled angiogenic molecules delivery can be achieved with
binary alginate
Blood vessel formation and perfusion can be significantly improved by using binary
alginate as local dual delivery vehicle
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FUTURE WORKVEGF 121 AND VEGF 165 PLAY DISTINCT ROLES IN THE STIMULATION OF COLLATERAL VESSEL AND BLOOD FLOW RE-ESTABLISHMENT IN AN ISCHEMIC HEART
VEGF 121
µmµm
VEGF 165
-+
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ACKNOWLEDGMENTSProfessor David J. Mooney
Professor Mário Barbosa
Department of RadiologyDivision of Cardiovascular DiseasesDepartment of Internal MedicineUniversity of Michigan
Paul M. Grossman
Sanjay Rajagopalan
Qinghua Sun
Mooney Lab
2nd PGDB
FCT - Fundação para a Ciência e a Tecnologia
IGC/FCG - Fundação Calouste Gulbenkian
NIH - National Institutes of Health
Harvard University/University of Michigan & Universidade do Porto
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Thinking about Curia 2005
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A - Alginate; B - New tissue; C - Blood Vessel; D - Inflammatory cells
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