Index
Abstract 1
Chapter 1: Introduction 7
1.1 Nanoscale and nanomaterials 8
1.2 Carbon nanotubes 11
1.3 Open issue: CNTs toxicity 12
1.4 Properties of CNTs exploited for biomedical applications: the future of
Nanomedicine 13
Chapter 2: Material and Methods 17
2.1 MWCNT preparation. 17
2.2 Preparation of MWCNTs functionalized with GDC-0941 19
2.3 Cell line culture 19
2.4 SH-SY5Y and HN9.10e preparation for the experiments of in vitro
magnetic translocation mediated by MWCNTs 21
2.5 PC12 preparation for the experiments of in vitro magnetic translocation
mediated by MWCNTs 21
2.6 Primary cells 22
2.6.1 MSC isolation and culture 22
2.6.2 Immunophenotyping and differentiation of MSC and MSC-CNT sample 23
2.6.3 Neurons culture 23
2.7 Biocompatibility assays 24
2.7.1 Trypan blue assay 24
2.7.3 Short-term assays 24
2.7.4 Long-term assays 25
2.7.5 Dose response curve 25
2.7.6 Oxidative stress evaluation 25
2.7.7 Cell staining and evaluation of apoptotic cell death 26
2.8 Flow cytometry 27
2.9 Staining of cell cytoskeletal actin 27
2.10 Sample preparation for FIB imaging 27
2.11 Preparation of samples for MRI 28
2.11.1 Micro-MRI 28 2.12 Western Blotting 29 2.13 Statistical analysis 30 2.14 In vivo experiments 30 2.14.1 MSC-CNT cells transplantation 30 2.14.2 Intracerebral injection of MWCTNs 31 2.15 Histological procedure 32 Chapter 3: Results 32 3.1 Biocompatibility of MWCNTs on SH-SY5Y 32
3.2 Biocompatibility of MWCNTs on NIH-3T3-L1 and effect on their
differentiation in adipocytes 40
3.3 Investigation of MWCNTs interaction with fibroblast cells 44
3.4 Biocompatibility of MWCNTs on PC-12 cells 46
3.5 Cell displacement mediated by magnetic carbon nanotubes: in vitro
demonstration in SH-SY5Y, HN9.10e and PC12 cells 49
3.6 Effects of Pluronic-coated carbon nanotubes in cortical neurons in vitro and
in vivo 54
3.6.2 PF127-MWCNTs solution does not induce damage in mouse brain 55
3.7 In vitro and in vivo effects of MWCNTs in Mesenchymal Stem Cells 57
3.8 Exploitation of the magnetic properties of MWCNTs: application for MSC
displacement in vitro 58
3.9 In-vivo study: guiding of the homing of mesenchymal stem cells after their
transplantation in rats by magnetic field 59
3.10 In vitro tracking of MSCs loaded with MWCNTs by using the Magnetic
Resonance Imaging (MRI) 64
3.10.1 MRI of MWCNTs 64
3.10.2 Stem cells tracking 64
3.11 Design and Develop of magnetic MWCNTs as carrier for the guided delivery of the innovative chemoterapic agent GDC-0941 produced by Astra
Zeneca in collaboration with the Life Science Dept of the University of Dundee 68
Chapter 4: 4. Discussion and conclusion 73
4.1 Biocompatibility of MWCNTs on SH-SY5Y 73
4.2 Biocompatibility of MWCNTs on NIH-3T3-L1 and effect on their
differentiation in adipocytes 74
4.3 Investigation of MWCNTs interaction with fibroblast cells 75
4.4 MWCNTs and PC-12 cells: can nanosciensce help neuroscience? 76
4.5 Exploitation of the magnetic properties of MWCNTs: application for cell
displacement in vitro of SH-SY5Y, HN9.10e and PC12 cells 77
4.6 Can MWCNTs compromise the viability of cortical neurons? In vitro and in
vivo studies 78
4.7 Exploitation of magnetic MWCNTs for stem cell displacement in vitro; driving of the homing of mesenchymal stem cells after their transplantation
4.8 Tracking of the cells loaded with nanoparticles by using the Magnetic
Resonance Imaging (MRI) 82
4.9 MWCNTs as carrier for the guided delivery of the innovative chemoterapic
agent GDC-0941 84 4.10 Concluding remark 86 References 87 Acknowledgements 97
