Toxicological evaluation of poly(ethyleneimine)-based non-viral vector systems for pulmonary siRNA application [Elektronische Ressource] / vorgelegt von Christiane Andrea Barbara Beyerle

Andy Adams

Sujets

• Naturwissenschaften

Informations

• Publié par: philipps-universitat_marburg
• Publié le: 01 janvier 2010
• Nombre de lectures: 19
• Langue: Deutsch
• Poids de l'ouvrage: 10 Mo

Extrait

Toxicological Evaluation of Poly(ethylene
imine) -based non-viral vector systems for
pulmonary siRNA application












Dissertation

zur Erlangung des Doktorgrades

der Naturwissenschaften

(Dr. rer. nat.)



dem Fachbereich Pharmazie der Philipps Universität Marburg vorgelegt von

Christiane Andrea Barbara Beyerle

aus Neuss







Marburg/Lahn 2010










































Vom Fachbereich Pharmazie der Philipps-Universität Marburg als Dissertation am
13.04.2010 angenommen.

Erstgutachter: Prof. Dr. Thomas Kissel

Zweitgutachter: Prof. Dr. Carsten Culmsee

Tag der mündlichen Prüfung am 14.04.2010
Die vorliegende Arbeit entstand auf Anregung und unter Anleitung von
Herrn Prof. Dr. Thomas Kissel

am Institut für Pharmazeutische Technologie und Biopharmazie
der Philipps-Universität Marburg


und in enger Zusammenarbeit mit der Arbeitsgruppe von
Herrn Dr. Tobias Stöger

am Institut für Lungenbiologie
Comprehensive Pneumology Center

im Helmholtz Zentrum München




































„Es geht darum, all das, was man zum Ausdruck bringen will, lange
und intensiv genug zu betrachten, um etwas daran zu entdecken, das
noch von niemandem gesehen und ausgesprochen worden ist. In allem
gibt es etwas Unbekanntes, denn wir sind daran gewöhnt, uns beim
Schauen stets an das zu erinnern, was man vor uns über das
Betrachtete gedacht hat.“ Guy de Maupassant, Reisen nach Nordafrika











Meinen Eltern und Großeltern
in Liebe und Dankbarkeit

Table of content

1 Introduction.......................................................................................... 1
The lung.................................................................................................................... 2
1.1 Anatomical and physiological aspects ........................................................... 2
1.2 Basic considerations for lung delivery........................................................... 7
1.3 Cellular Transport mechanisms................................................................... 13
1.4 Barriers for pulmonary administration........................................................ 17
1.5 Non viral vector systems for pulmonary siRNA application........................ 19
1.6 Aspects of Nanotoxicology in Nanomedicine ............................................... 25
1.7 Aim of this thesis .......................................................................................... 30
1.8 References .................................................................................................... 32
2 In vitro cytotoxic and immuno-modulatory profiling of low
molecular weight polyethylen-imines for pulmonary application 38
2.1 Abstract ........................................................................................................ 39
2.2 Introduction.................................................................................................. 40
2.3 Materials and Methods................................................................................. 43
2.4 Results .......................................................................................................... 48
2.5 Discussion .................................................................................................... 57
2.6 Supplementary material ............................................................................... 62
2.7 References .................................................................................................... 67
3 PEGylation affects cytotoxicity and cell-compatibility of
Poly(ethylene imine) for lung application: structure-function-
relationships .......................................................................................69
3.1 Abstract ........................................................................................................ 70
3.2 Introduction.................................................................................................. 71
3.3 Materials and Methods................................................................................. 74
3.4 Results .......................................................................................................... 80
3.5 Discussion .................................................................................................... 90
3.6 Supplementary material ............................................................................... 95
3.7 References .................................................................................................... 96
4 Toxicity pathway focused gene expression profiling of PEI-based
polymers for pulmonary applications............................................100
4.1 Abstract ...................................................................................................... 101
4.2 Introduction................................................................................................ 102
4.3 Experimental Section.................................................................................. 105
4.4 Results ........................................................................................................ 110
4.5 Discussion .................................................................................................. 118
4.6 Supporting Information.............................................................................. 124
4.7 References .................................................................................................. 130
5 Investigations on mutant frequency induced by PEI in FE1-
Muta Mouse lung epithelial cells.................................................134 
5.1 Abstract ...................................................................................................... 135
5.2 Introduction................................................................................................ 136
5.3 Materials and Methods............................................................................... 138
5.4 Results and Discussion............................................................................... 141
5.5 Conclusion.................................................................................................. 147
5.6 References .................................................................................................. 148

6 Side-effects of PEI-based siRNA nanocarriers for pulmonary
application in mice...........................................................................150
6.1 Abstract ...................................................................................................... 151
6.2 Introduction................................................................................................ 152
6.3 Materials and Methods............................................................................... 154
6.4 Results ........................................................................................................ 157
6.5 Discussion .................................................................................................. 168
6.6 Supplementary Material............................................................................. 174
6.7 References .................................................................................................. 183
7 Fatty acid modification of low molecular weight polyethylenimine-
mediated siRNA delivery to lung leucocytes after intratracheal
instillation in mice............................................................................185
7.1 Abstract ...................................................................................................... 186
7.2 Background ................................................................................................ 187
7.3 Methods ...................................................................................................... 189
7.4 Results ........................................................................................................ 194
7.5 Discussion .................................................................................................. 205
7.6 Supplementary data.................................................................................... 208
7.7 References .................................................................................................. 211
8 Summary and Perspectives.............................................................214
8.1 Summary................................................