Photophysical characterization and optimization of novel polymer based photosensitizer carrier systems for PDT [Elektronische Ressource] / von Kuan Chen

Kuan Chen

Sujets

• Physik
• Gesundheit

Informations

• Publié par: humboldt-universitat_zu_berlin
• Publié le: 01 janvier 2010
• Nombre de lectures: 7
• Langue: English
• Poids de l'ouvrage: 1 Mo

Extrait

Photophysical characterization and optimization of novel polymer based
photosensitizer carrier systems for PDT

Dissertation
zur Erlangung des akademischen Grades
doctor rerum naturalium
( Dr. rer. nat.)
im Fach (Physik)
eingereicht an der

Mathematisch-Naturwissenschaftlichen Fakultät I
der Humboldt-Universität zu Berlin
von

M. Sc. Kuan Chen
geb. am 27.10.1979 in Beijing, China

Präsident der Humboldt Universität zu Berlin
Prof. Dr. Christoph Markschies

Dekan der Mathematisch-Naturwissenschaftlichen Fakultät I
Prof. Dr. Lutz-Helmut Schön

Gutachter/innen: 1. Prof. Dr. Beate Röder
2. Prof. Dr. Jürgen Rabe
3. Prof. Dr. Matthias Ballauf
4. Prof. Dr. Karsten Heyne

Tag der mündlichen Prüfung: 22.06.2010 Table of contents
Table of contents
List of symbols and abbreviations…………………………………………………...4
0. Introduction..............................................................................................................6
1. Brief history of Photodynamic Therapy and fundamentals of
Photosensitization………………….........................................................................…9
1.1 Brief history of Photodynamic Therapy...............................................................9
1.2 Molecular mechanism of photosensitization........................................................10
1.2.1 Photosensitization………….………..……….…….………...……………10
1.2.2 Deactivation of singlet oxygen via radiative way............................................11
1.2.3 Singlet oxygen quantum yield..........................................................................12
1.2.4 Singlet oxygen detection in cells.....................................................................13
1.3 Electronic excited states of molecular oxygen.....................................................16
1.4 Requirements of an efficient photosensitizer …….................... ... .......…….….17
2. Tumour targeting and nanoparticle drug carrier system...................................22
2.1 Passive strategies of tumour targeting..................................................................22
2.2 Active strategies of tumour targeting...................................................................23
2.3 Nanoparticle as drug carrier system.....................................................................24
2.4 Biodegradable nanoparticles................................................................................25
3. Methods...................................................................................................................27
3.1 Ground state absorption (UV-vis spectra)...........................................................27
3.2 Steady-state fluorescence………………….…………………………………27
3.3 TCSPC……………………………….…………………………………………27
3.3.1 Fluorescence lifetime……………………………………………………...27
3.3.2 Decay associated fluorescence spectra (DAFS)…………………...……...29
3.4 Time-resolved singlet oxygen luminescence……………………...………….29
3.4.1 Singlet oxygen luminescence detection in solution……………………….30
3.4.2 Singlet oxygen luminescence detection in living cells………………….....31
3.5 Laser flash photolysis…………………………………………...……………32
3.6 Cell experiment…………………………………………...………………….34
1Table of contents
4. Materials and basic experiments………………………………………………...35
4.1 Photosensitizers………………………………………...……………………….35
4.2 HSA nanoparticles……………………………………………………………....36
4.3 PLGA nanoparticles……………………………………………………...……..37
4.4 Pheo-HSA nanoparticles as starting system…………………………………….38
4.4.1 Preparation of Pheo-HSA nanoparticles……………………...……………..38
4.4.2 Photophysical characterization of Pheo-HSA nanoparticles………………..39
4.5 Photo-induced singlet oxygen generation in cells……………………………....44
4.5.1 Laser flash photolysis……………………………………...………………...44
4.5.2 Intracellular singlet oxygen luminescence……………………………...…...45
4.6 Conclusions………………………………………………………...…………...46
5. HSA-nanoparticles as biodegradable carrier system for PDT………………...48
5.1 Photophysical properties of mTHPP-HSA with different loading ratios………..48
5.1.1 Sample description……………………………………………………...…...49
5.1.2 Absorption and steady-state fluorescence……………...……………………49
5.1.3 Fluorescence kinetic parameters………………………………………….52
5.1.4 Singlet oxygen generation……………………………………..…………….54
5.2 Photophysical properties of mTHPC-HSA with different loading ratios……….56
5.2.1 Sample description…………………………………………………………..56
5.2.2 Absorption spectra…………………………………………………….…….57
5.2.3 Steady-state fluorescence spectra……………………………………...…….60
5.2.4 Fluorescence kinetic parameters……………….…………………62
5.2.5 Singlet oxygen generation and triplet lifetime in solution……………...…...63
5.3 Photophysical investigations of mTHPC-HSA with different cross linkages….66
5.3.1 Absorption and steady-state fluorescence………………………….………..67
5.3.2 Fluorescence kinetic parameters…………………………………….……68
5.3.3 Singlet oxygen generation and laser flash photolysis……….………………69
5.3.4 Singlet oxygen luminescence detection in Jurkat cells……………………...70
5.4 Conclusions…………………………………………………….……………….72
6. PLGA -nanoparticles as biodegradable carrier system for PDT………..…….75
2Table of contents
6.1 Comparison to the photophysical properties of mTHPP- and mTHPC-PLGA…75
6.2 Absorption and steady-state fluorescence…………………………………..…..75
6.3 Fluorescence kinetic parameters and DAFS…………………...……………….78
6.4 Singlet oxygen generation and triplet lifetime………………………………….82
6.5 PLGA matrix decomposing in organic solvent ...………………………………84
6.6 PLGA matrix decomposition and drug release in Jurkat cells …....……………87
6.7 Conclusions……………………………………………………………..………90
7. Comparison between PLGA and HSA nanoparticles……………………….…91
7.1 Singlet oxygen generation of mTHPP-PLGA and mTHPP-HSA in solution…...91
7.2 Behaviour of mTHPP-PLGA and mTHPP-HSA nanoparticles in Jurkat cells.....92
7.3 Conclusions……………………………………………….…………………….94
8. PLGA- and HSA- NPs as carrier for active tumour targeting……………….95
8.1 Covalently conjugated and antibody modified mTHPC-HSA nanoparticles…...95
8.1.1 Sample description……………………………………………………….….96
8.1.2 Absorption and steady-state fluorescence……………………………….…..96
8.1.3 Fluorescence kinetic parameters……………………………………….……97
8.1.4 Singlet oxygen generation and laser flash photolysis……………………….98
8.1.5 Singlet oxygen luminescence detection in cells……………..………...……..99
8.2 Investigation on mTHPC-PLGA nanoparticles modified with antibodies….....100
8.2.1 Sample description……………………………………………..…………..101
8.2.2 Absorption and steady-state fluorescence……………………………….…101
8.2.3 Fluorescence kinetic parameters………………………………...…………102
8.2.4 Singlet oxygen generation and laser flash photolysis………………...……103
8.2.5 Singlet oxygen luminescence in HT-29 cells………………………….……103
8.3 Conclusions……………………………………………………………………104
9. Summary/Zusammenfassung…………………..………………………………106
Reference……………………………………………………………………...……113
Acknowledgement………………………………….………………………...……129
List of Publications…………………………...……….……………………...……130
3List of symbols and abbreviations
List of symbols and abbreviations

CePC-HSA Cetuximab modified mTHPC-HSA nanoparticles
CNPL1, 2, 3 mTHPC-PLGA nanoparticles with different loading ratios
CoPC-HSA Covalently binding mTHPC-HSA nanoparticles
D O Deuterium oxide 2
f Energy transfer efficiency factor ET
FWHM Full width at half maximum
HpD Hematoporphyrin Derivates
HSA Human Serum Albumin nanoparticles
HLPP-HSA High loading ratio mTHPP-HSA nanoparticles
IC Internal Conversion
IgGPC-HSA IgG modified mTHPC-HSA nanoparticles
ISC Inter-System Crossing
LLPP-HSA Low loading ratio mTHPP-HSA nanoparticles
Mol, M Molar, Molar / Liter
mTHPC 5, 10, 15, 20-Tetrakis (3-hydroxyphenyl)-chlorin
mTHPC-PLGA mTHPC loaded PLGA nanoparticles
mTHPP 5, 10, 15, 20-Tetrakis (3-hydroxyphenyl)-porphyrin
mTHPP-COOH 5-(4-Carboxyphenyl)-10, 15, 20-tris (3-hydroxyphenyl)-
porphyrin
mTHPP-PLGA mTHPP loaded PLGA nanoparticles
NP Nanoparticle
NPC mTHPC-HSA nanoparticles with different cross-linkages
NPL1, 2, 3, 4, 5 mTHPC-HSA nanoparticles with different loading ratios
OD Optical Density
OPO / OPA Optical Parameter Oscillator / Optical Parameter Amplifier
PDT Photodynamic Therapy
PEG Polyethylene glycol
4List of symbols and abbreviations
Pheo P