77. Magnetic Nanoparticles For Cancer

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77. Magnetic Nanoparticles For Cancer

 

 

CATEGORY:Alternative Medicines 300 Courses

COURSE NUMBER: 77

FEES: 555/- INR only

CERTIFICATE VALIDITY: Lifetime

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BOOKS/ MANUALS: Pages

Syllabus

ACKNOWLEDGEMENTS………………………………………………………………………………………………iii
Table of Contents……………………………………………………………………………………………………………..v
List of Tables………………………………………………………………………………………………………………….ix
List of Figures………………………………………………………………………………………………………………….x
Chapter 1 Introduction ………………………………………………………………………………………………………1
1.1 Objectives …………………………………………………………………………………………………………3
Chapter 2 Active targeting strategies of iron oxide magnetic nanoparticles………………………………5
2.1 Introduction…………………………………………………………………………………………………………….5
2.2 Active Targeting ……………………………………………………………………………………………………..8
2.2.1 Antibodies………………………………………………………………………………………………………10
2.2.2 Aptamers………………………………………………………………………………………………………..14
2.2.3 Peptides………………………………………………………………………………………………………….17
2.2.4 Small Molecules………………………………………………………………………………………………25
2.3 Conclusions and Future Perspectives………………………………………………………………………..31
Chapter 3 Magnetic nanoparticles and nanocomposites for remote controlled therapies…………..33
3.1 Introduction…………………………………………………………………………………………………………..33
3.2 Remote controlled energy release…………………………………………………………………………….36
3.2.1 Thermal therapy………………………………………………………………………………………………36
3.2.2 Nanoscale Energy Delivery ………………………………………………………………………………42
3.3 Combination therapy………………………………………………………………………………………………47
3.4 Conclusions…………………………………………………………………………………………………………..53
Chapter 4 Synthesis and characterization of CREKA-conjugated iron oxide nanoparticles for
hyperthermia applications………………………………………………………………………………………………..55
4.0 Abstract………………………………………………………………………………………………………………..55
4.1 Introduction…………………………………………………………………………………………………………..56
4.2 Materials and Methods……………………………………………………………………………………………59
4.2.1 Materials ………………………………………………………………………………………………………..59
4.2.2 Crosslinked dextran coated iron oxide nanoparticle synthesis ……………………………….60
4.2.3 CREKA conjugation of iron oxide nanoparticles …………………………………………………60
4.2.4 Particle characterization……………………………………………………………………………………62
4.2.5 Cytotoxicity analysis of particle systems…………………………………………………………….64
4.2.6 CREKA and CREKA IONP binding affinity to fibrinogen clots ……………………………64
4.2.7 In vitro magnetically mediated hyperthermia and cisplatin combined treatment ………65
4.2.8 Statistical analysis……………………………………………………………………………………………66
4.3 Results and Discussion …………………………………………………………………………………………..66
4.3.1 Particle characterization……………………………………………………………………………………66
4.3.2 Remote controlled heating of iron oxide magnetic nanoparticles via AMF ……………..69
4.3.3 Cytotoxicity analysis of particles……………………………………………………………………….70
4.3.4 Fibrinogen binding affinity ……………………………………………………………………………….71
4.3.5 Magnetically-mediated hyperthermia …………………………………………………………………72
4.4 Conclusion ……………………………………………………………………………………………………………76
Chapter 5 The effects of synthesis method on the physical and chemical properties of dextran
coated iron oxide nanoparticles ………………………………………………………………………………………..78
5.0 Abstract………………………………………………………………………………………………………………..78
5.1 Introduction…………………………………………………………………………………………………………..79
5.2 Materials and Methods……………………………………………………………………………………………83
5.2.1 Materials ………………………………………………………………………………………………………..83
5.2.2 Dextran coated iron oxide nanoparticle synthesis via two-step method …………………..84
5.2.3 Dextran coated iron oxide nanoparticle synthesis via semi-two-step method …………..84
5.2.4 Dextran coated iron oxide nanoparticle synthesis via simultaneous semi-two-step
method …………………………………………………………………………………………………………………..85
5.2.5 Dextran coated iron oxide nanoparticle synthesis via one-step method …………………..85
5.2.6 Epichlorohydrin crosslinking of dextran coated iron oxide nanoparticles………………..85
5.2.7 Particle characterization……………………………………………………………………………………86
5.3 Results and Discussion …………………………………………………………………………………………..88
5.4 Conclusions…………………………………………………………………………………………………………..99
Chapter 6 Peptide conjugated magnetic nanoparticles for magnetically mediated energy delivery
…………………………………………………………………………………………………………………………………..101
6.0 Abstract………………………………………………………………………………………………………………101
6.1 Introduction…………………………………………………………………………………………………………102
6.2 Materials and Methods………………………………………………………………………………………….107
6.2.1 Materials ………………………………………………………………………………………………………107
6.2.2 Synthesis of uncoated iron oxide nanoparticles………………………………………………….107
6.2.3 Synthesis of dextran coated iron oxide nanoparticles (Fe3O4 + Dx) ………………………108
6.2.4 Epichlorohydrin (ECH) crosslinking of dextran coated iron oxide nanoparticles (Fe3O4
+ Dx-ECH)……………………………………………………………………………………………………………108
6.2.5 Amine functionalization of crosslinked dextran coated iron oxide nanoparticles (Fe3O4
+ Dx-ECH-Amine)…………………………………………………………………………………………………108

6.2.6 TAT peptide conjugation to iron oxide nanoparticles (Fe3O4 + TAT)……………………109
6.2.7 Iron concentration assay …………………………………………………………………………………109
6.2.8 Iron oxide nanoparticle characterization……………………………………………………………109
6.2.9 Cell Culture…………………………………………………………………………………………………..110
6.2.10 Iron oxide nanoparticle uptake……………………………………………………………………….110
6.2.11 Reactive Oxygen Species (ROS) Generation……………………………………………………111
6.2.12 Acridine orange lysosomal permeabilization experiments…………………………………111
6.2.13 Mitochondrial membrane depolarization studies (JC-1)…………………………………….112
6.2.14 Caspase 3/7 apoptosis studies………………………………………………………………………..112
6.2.15 Viability studies…………………………………………………………………………………………..112
6.3 Results and Discussion …………………………………………………………………………………………113
6.3.1 Iron oxide nanoparticle characterization……………………………………………………………113
6.3.2 Cellular uptake of iron oxide nanoparticles……………………………………………………….116
6.3.3 Intracellular reactive oxygen species formation …………………………………………………117
6.4 Conclusions…………………………………………………………………………………………………………125
Chapter 7 Targeted iron oxide nanoparticles for the enhancement of radiation therapy ………….127
7.0 Abstract………………………………………………………………………………………………………………127
7.1 Introduction…………………………………………………………………………………………………………128
7.2 Materials and Methods………………………………………………………………………………………….133
7.2.1 Materials ………………………………………………………………………………………………………133
7.2.2 Synthesis of uncoated iron oxide nanoparticles (uncoated Fe3O4)…………………………133
7.2.3 Synthesis of dextran coated iron oxide nanoparticles (Fe3O4 + Dx) ………………………134
7.2.4 Epichlorohydrin (ECH) crosslinking of dextran coated iron oxide nanoparticles (Fe3O4
+ Dx-ECH)……………………………………………………………………………………………………………134
7.2.5 Amine functionalization of crosslinked dextran coated iron oxide nanoparticles (Fe3O4
+ Dx-ECH-Amine)…………………………………………………………………………………………………134
7.2.6 TAT peptide conjugation to iron oxide nanoparticles (Fe3O4 + TAT)……………………135
7.2.7 Iron Concentration Assay………………………………………………………………………………..135
7.2.8 Cell Culture…………………………………………………………………………………………………..135
7.2.9 Transmission electron microscopy……………………………………………………………………135
7.2.10 Acridine orange lysosomal permeabilization experiments…………………………………136
7.2.11 Mitochondrial stress test ……………………………………………………………………………….136
7.2.12 Reactive Oxygen Species (ROS) Generation……………………………………………………138
7.2.13 Viability studies…………………………………………………………………………………………..138
7.3 Results and Discussion …………………………………………………………………………………………138
7.4 Conclusions…………………………………………………………………………………………………………149
Chapter 8 Conclusions…………………………………………………………………………………………………..151
8.1 Significant Findings……………………………………………………………………………………………..152
Appendix A Combination treatment of multi-cellular tumor spheroids………………………………..154
A. 1 Introduction……………………………………………………………………………………………………….154
A.2 Materials and Methods…………………………………………………………………………………………158
A.2.1 Materials………………………………………………………………………………………………………158
A.2.2 Cell culture…………………………………………………………………………………………………..159
A.2.3 MCTS formation…………………………………………………………………………………………..159
A.2.4 MCTS dissociation………………………………………………………………………………………..160
A.2.5 Acid phosphatase assay………………………………………………………………………………….160
A.2.6 Resazurin assay…………………………………………………………………………………………….160
A.2.7 Calcein AM/Ethidium homodimer-1 (live/dead) assay ………………………………………161
A.2.8 Assay validation studies…………………………………………………………………………………161
A.2.9 Incubator-induced hyperthermia with CDDP ……………………………………………………162
A.2.10 Incubator induced hyperthermia with radiation ……………………………………………….163
A.3 Results and discussion …………………………………………………………………………………………163
A.3.1 MCTS assay validation ………………………………………………………………………………….163
A.3.2 Cisplatin combined with hyperthermia on MCTS ……………………………………………..168
A.3.3 Radiation combined with hyperthermia on MCTS …………………………………………….169
A.4 Conclusions………………………………………………………………………………………………………..173
References……………………………………………………………………………………………………………………175
Vita……………………………………………………………………………………………………………………………..202

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