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Table of contents
1. Chapter I: State of art
1.1 General Introduction
1.2 Ligament structure
1.2.1 Gross structure
1.2.2 Microscopic organization
1.3 Ligament composition
1.3.1 Cellular component
1.3.2 Molecular components
1.4 Ligament function
1.5 Ligament biomechanics
1.6 Ligament injuries
1.6.1 Epidemiology of ligament injuries
1.6.2 Ligaments response to injury
1.6.3 Solutions for ligament repair
1.7 Ligament tissue engineering
1.7.1 Requirements for defining scaffolds for ligament tissue engineering
1.7.2 Biomaterial resources
1.7.3 Scaffold design
1.7.4 Cell sources for ligament tissue engineering
1.8 Biomechanical/Biochemical stimulation
1.8.1 Biochemical stimulation
1.8.2 Biomechanical stimulation with bioreactors
1.9 Conclusion
1.9.1 Summary, issues and hypothesis
Objective and content of the thesis
2 Chapter II : Materials and methods
2.1 Biological study of MSCs
2.1.1 Isolation and expansion of BM-MSCs and WJ-MSCs
2.1.2 Colony Forming Unit-fibroblast (CFU-F)
2.1.4 Differentiation
2.1.5 Senescence
2.2 Scaffold fabrication
2.2.1 PLCL multilayer braided scaffolds
2.2.2 Silk and silk/PLCL multilayer braided scaffolds
2.3 Scaffold modification with Layer-By-Layer (LBL) technology
2.3.1 Scaffold with PLL/HA modification
2.3.2 Fibers modification with PLL-FITC/HA
2.4 Scaffold characterization
2.4.1 Structure by Fourier transform infrared spectroscopy (FTIR)
2.4.2 Morphology by Scanning Electronic microscopy (SEM)
2.4.3 Morphology by confocal laser macroscopy (CLSM)
2.4.4 Topology by atomic force microscopy (AFM)
2.4.5 Mechanical properties
2.4.6 Morphology and Porosity by micro computed tomography (μCT)
2.5 Evaluation of biocompatibility of MSCs on scaffolds
2.5.1 Scaffold sterilization
2.5.2 Cell seeding on the scaffold
2.5.3 Cell proliferation evaluated by AB test
2.5.4 Cell location and morphology detection by SEM
2.5.5 Cell morphology observation by CLSM and fluorescent microscopy
2.5.6 Live/dead staining of MSCs on scaffolds
2.5.7 Detection of Extracellular matrix synthesis by fluorescent microscopy
2.5.8 Cell migration stimulated by scaffolds
2.5.9 Histology and immunohistochemistry of MSCs on scaffolds
2.5.10 Bioreactor sterilization and assembly
2.5.11 MSCs seeded on scaffolds with mechanical stimulation and biological evaluation
2.6 Conclusion of the present chapter
3 Chapter III : Biological study of cell sources selected for ligament tissue engineering
3.1 Introduction
3.2 Results and discussions
3.2.1 Biological characteristics of WJ-MSCs and BM-MSCs
3.3 Conclusion
4 Chapter IV : Surface modification of a braided multilayer PLCL scaffold for ligament tissue engineering
4.1 Introduction
4.2 Results and discussion
4.2.1 Physicochemical properties of PLCL
4.2.2 Biocompatibility of MSCs on PLCL scaffold
4.3 Conclusion
5 Chapter V : Proposition of silk and silk/PLCL scaffold for ligament tissue engineering
5.1 Introduction
5.2 Results and discussion
5.2.1 Physiochemical properties of silk-based braided scaffold
5.2.2 Biological properties of silk-based braided scaffold
6 Chapter VI : An attempt to study effect of dynamic mechanical stimulation on MSCs-construct in a tension-torsion bioreactor
6.1 Introduction
6.2 Results and discussion
6.2.1 MSCs metabolic activity of MSCs on scaffold
6.2.2 MSCs morphology and location on scaffolds
6.3 Conclusion
7 Chapter VII: Discussion
7.1 Initial PLCL braided scaffold
7.2 PLL/HA LBL modification
7.3 Limitations of the initial PLCL braided scaffold
7.4 Silk as an alternative to PLCL
7.5 Proposition of a silk-based braided scaffold
7.7 Preliminary study of the effect of mechanical stimulation on MSC-scaffold differentiation
8 Chapter VIII: Conclusion, limitation and perspectives
8.1 Conclusion of the present work
8.2 Limitations of the proposed scaffolds and required further characterization
8.2.1 Biodegradation properties
8.2.2 In vivo implantation
8.2.3 Quantitative characterization
8.3 Perspectives of the present work
References
