Contents
Abstract
Index
1. Introduction
1.1 Tissue engineering1.1.1 Cardiovascular tissue engineering
1.2 Cell sources
1.2.1 Endothelial Progenitor Cells (EPCs) 1.2.2 EPCs in tissue engineering
1.3 Scaffolds
1.3.1 Scaffolds for cardiovascular applications 1.3.2 Fibrin
1.3.3 Peptide amphiphiles (PA) 1.3.4 RGD-containing PA
2. Aim
3. Materials and Methods
3.1 Endothelial progenitor cells (EPCs) culture
3.2 Human umbilical vein endothelial cells (HUVECs) isolation and culture 3.3 EPC characterization
3.3.2 Immunocytochemistry 3.3.3 Cell proliferation
3.3.4 In vitro angiogenic assay
3.4 Scaffold preparation
3.4.1 Fibrin gels
3.4.2 Peptide amphiphile (PA)
3.4.2.1 PA synthesis 3.4.2.2 Gel formation
3.5 Scaffold ultrastructure
3.5.1 Scanning Electron Microscopy (SEM)
3.5.2 Cryo-Scanning Electron Microscopy (Cryo-SEM) 3.5.3 Atomic Force Microscopy (AFM)
3.6 EPC culture and characterization on biomaterials
3.6.1 Cell viability
3.6.2 Flow cytometry analysis
3.6.3Confocal Laser Scanning Microscopy 3.6.4 EPC angiogenic activity
3.6.4.1 Matrigel assay 3.6.4.2 Cytokine release
3.6.5 Stem cell markers expression
3.6.5.2 Real Time PCR (Quantitative PCR, QPCR)
3.7 Data presentation and statistical analysis
4. Results
4.1 EPC culture and characterization on fibronectin
4.1.1 Endothelial markers expression 4.1.2 Cell proliferation
4.1.3 In vitro angiogenesis assay
4.2 Scaffold formation
4.2.1 Effect of fibrin composition on scaffold stiffness
4.2.2 Effect of fibrin composition on fibre diameter and density 4.2.3 PA self-supporting gel formation
4.2.4 Effect of CaCl2 concentration on PA stiffness
4.3 Scaffold microarchitecture
4.3.1 Fibrin 4.3.2 PA
4.4 EPC culture and characterization on scaffolds
4.4.1 EPCs on fibrin
4.4.1.2 Cell viability
4.4.1.2 EPC differentiation
4.4.1.3 Expression of stemness markers 4.4.1.4 In vitro angiogenesis assay 4.4.1.5 Cytokine release
4.4.2 EPCs on PA
4.4.1.1 Cell viability
4.4.1.2 EPC differentiation 4.4.1.3 Cytokine release
5. Discussion
5.1 Fibrin and EPCs 5.2 PA and EPCs 5.3 Limitations5.4. Future developments
