VEGF carrying coatings for enhanced vascular graft endothelialization:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
München
Verl. Dr. Hut
2010
|
| Ausgabe: | 1. Aufl. |
| Schriftenreihe: | Pharmazeutische Technologie
|
| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-125453 Inhaltsverzeichnis |
| Beschreibung: | 169 S. Ill., graph. Darst. |
| ISBN: | 9783868537567 |
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| 245 | 1 | 0 | |a VEGF carrying coatings for enhanced vascular graft endothelialization |c Frank Schaubhut |
| 250 | |a 1. Aufl. | ||
| 264 | 1 | |a München |b Verl. Dr. Hut |c 2010 | |
| 300 | |a 169 S. |b Ill., graph. Darst. | ||
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Datensatz im Suchindex
| _version_ | 1804143765395341312 |
|---|---|
| adam_text | IMAGE 1
TABLE OF CONTENTS
CHAPTER 1
INTRODUCTION AND OBJECTIVE OF THE THESIS
1 INTRODUCTION 2
2 VASCULAR GRAFTS 3
2.1 MATERIALS USED FOR VASCULAR GRAFTS 3
2.1.1 DACRON 4
2.1.2 EPTFE 4
2.2 FAILURE OF VASCULAR GRAFTS 5
2.3 IMPROVEMENT OF VASCULAR GRAFT PERFORMANCE 6
3 ENDOTHELIAL CELLS AND THEIR ROLE IN VESSEL REPLACEMENT 7
3.1 ENDOTHELIAL CELL RELATED PROBLEMS 8
3.2 APPROACHES FOR ENDOTHELIALIZATION 8
3.2.1 IN VITRO ENDOTHELIALIZATION 9
3.2.2 IN VIVO ENDOTHELIALIZATION 9
4 POLYMERS FOR CONTROLLED RELEASE APPLICATIONS 11
4.1 NATURAL POLYMERS - COLLAGEN 11
4.1.1 STRUCTURE OF COLLAGEN 12
4.1.2 CROSSLINKING OF COLLAGEN 13
4.2 SYNTHETIC POLYMERS - PLGA 14
5 CONCLUSIONS 16
6 OBJECTIVES OF THE THESIS 17
7 REFERENCES 19
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/100933039X
DIGITALISIERT DURCH
IMAGE 2
CHAPTER 2
MODIFICATION OF PTFE USING ATMOSPHERIC PLASMA AND ITS CHARACTERIZATION
27
1 INTRODUCTION 28
2 MATERIALS AND METHODS 29
2.1 MATERIALS 29
2.2 METHODS 29
2.2.1 PLASMA ACTIVATION 29
2.2.2 CHARACTERIZATION OF SURFACES AFTER PLASMA MODIFICATION 30
2.2.2.1 SCANNING ELECTRON MICROSCOPY 30
2.2.2.2 X-RAY PHOTOELECTRON SPECTROSCOPY (XPS) 30
2.2.2.3 ATTENUATED TOTAL REFLECTION- FT-IR SPECTROSCOPY (ATR-FT-IR) 30
2.2.2.4 SURFACE ENERGY DETERMINATION 30
2.2.3 CELL COMPATIBILITY 31
2.2.4 FILM ADHESION 31
3 RESULTS AND DISCUSSION 31
3.1 SURFACE CHARACTERIZATION 31
3.1.1 SURFACE MORPHOLOGY 32
3.1.2 SURFACE CHEMISTRY ANALYSIS BY ATR-FTIR 33
3.1.3 SURFACE COMPOSITION 34
3.1.4 DETERMINATION OF SURFACE ENERGY - HYDROPHILICITY 37
3.2 OPTIMIZATION OF THE PLASMA ACTIVATION PROCESS USING THE PLASMABRUSH
38
3.2.1 OPTIMIZATION OF IGNITION VOLTAGE 38
3.2.2 OPTIMIZATION OF TREATMENT TIME 40
3.2.3 INFLUENCE OF AGING TIME POST TREATMENT ON SURFACE ENERGY 41
3.3 CELL COMPATIBILITY OF PLASMA TREATED SURFACES 42
3.4 FILM ADHESION 43
4 CONCLUSIONS 45
5 REFERENCES 46
IMAGE 3
TABLE OF CONTENTS
CHAPTER 3
INVESTIGATION OF PLGA FILMS FOR CONTROLLED RELEASE APPLICATIONS 49
1 INTRODUCTION 50
2 MATERIALS AND METHODS 51
2.1 MATERIALS 51
2.2 METHODS 52
2.2.1 FITC-DEXTRAN SIZE REDUCTION 52
2.2.1.1 SWING MILL 52
2.2.1.2 SPRAY DRYING 52
2.2.2 MORPHOLOGICAL ANALYSIS 52
2.2.2.1 SCANNING ELECTRON MICROSCOPY 52
2.2.2.2 LASER DIFFRACTION 53
2.2.3 PLGA COATING PROCEDURE 53
2.2.4 IN-VITRA RELEASE STUDIES 53
2.2.5 CELL COMPATIBILITY TESTING 54
2.2.6 EVALUATION OF CELL QUANTIFICATION METHODS 54
2.2.6. F PROPIDIUM IODIDE DYE 54
2.2.6.2 FLUORESCENCE ACTIVATED CELL SORTING (FACS) 54
2.2.6.3 CELL TITER-BLUE. 55
3 RESULTS AND DISCUSSION 55
3.1 FITC-DEXTRAN SIZE REDUCTION 55
3.1.1 SIZE REDUCTION BY MILLING 55
3.1.2 SIZE REDUCTION USING SPRAY DRYING 56
3.2 IN VITRO RELEASE STUDIES 57
3.2.1 INFLUENCE OF PLGA MOLECULAR WEIGHT ON RELEASE RATES 58
3.2.2 INFLUENCE OF ESTERIFICATION ON RELEASE RATES 61
3.2.3 INFLUENCE OF FITC-DEXTRAN PARTICLE SIZE ON RELEASE RATES 63
3.3 CELL COMPATIBILITY OF PLGA COATINGS 64
3.4 EVALUATION OF CELL QUANTIFICATION METHODS 65
3.4.1 CELL QUANTIFICATION USING FACS 66
3.4.2 CELL QUANTIFICATION USING PROPIDIUM IODIDE DYE 67
3.4.3 CELL QUANTIFICATION USING A CELL COUNTING CHAMBER 68
3.4.4 CELL QUANTIFICATION USING CELL TITER-BLUE* 70
4 CONCLUSIONS 73
5 REFERENCES.. 74
IMAGE 4
TABLE OF CONTENTS
CHAPTER 4
INVESTIGATION OF COLLAGEN FILMS FOR CONTROLLED RELEASE OF VEGF 165 77
1 INTRODUCTION 78
2 MATERIALS AND METHODS 79
2.1 MATERIALS 79
2.2 METHODS 80
2.2.1 COLLAGEN CROSSLINKING 80
2.2.1.1 CROSSLINKING PROCEDURE 80
2.2.1.2 LYOPHILIZATION 80
2.2.1.3 DIFFERENTIAL SCANNING CALORIMETRY 81
2.2.1.4 KARL-FISCHER ORATION 81
2.2.2 COLLAGEN COATING PROCEDURE 82
2.2.2.1 PREPARATION OF COLLAGEN DISPERSION 82
2.2.2.2 COATING PROCEDURE - COVER COATING 82
2.2.2.3 COATING PROCEDURE -SQUEEGEE 83
2.2.3 CELL COMPATIBILITY 83
2.2.4 IN-VITRO RELEASE STUDIES 84
2.2.4.1 RELEASE STUDY OF FITC-DEXTRAN FROM COLLAGEN MATRICES 84
2.2.4.2 RELEASE STUDY OF VEGF T65 FROM COLLAGEN MATRICES 84
2.2.5 INFLUENCE ON CELL GROWTH BY INCORPORATED V E G F IM 85
2.2.5.1 RAT AORTIC SMOOTH MUSCLE CELLS. 85
2.2.5.2 CELL GROWTH STUDY. 85
3 R E S U L TS A ND D I S C U S S I ON 86
3.1 CHARACTERIZATION OF CROSSLINKED COLLAGEN 86
3.1.1 DIFFERENTIAL SCANNING CALORIMEFRY 86
3.1.2 RESIDUAL MOISTURE OF CROSSLINKED COLLAGEN 88
3.2 CELL COMPATIBILITY OF COLLAGEN VARIANTS 88
3.3 IN VITRO RELEASE STUDIES 90
3.3.1 INFLUENCE OF COATING PROCEDURE ON FITC-DEXTRAN RELEASE PROFILE 90
3.3.2 INFLUENCE OF CROSSLINKING ON FITC-DEXTRAN RELEASE PROFILE 92
3.3.3 RELEASE OF VEGF 165 FROM COLLAGEN MATRICES 94
3.4 INFLUENCE OF INCORPORATION OF VEGF 165 IN COLLAGEN MATRICES ON CELL
GROWTH 96
3.4.1 SMOOTH MUSCLE CELLS 97
3.4.2 VASCULAR ENDOTHELIAL CELLS 98
4 CONCLUSIONS 101
5 REFERENCES 103
IMAGE 5
TABLE OF CONTENTS
CHAPTER 5
CELL GROWTH STIMULATION BY VEGF 165 COVALENTLY LINKED TO A MODIFIED
PLGA-MATRIX 107
1 INTRODUCTION 108
2 MATERIALS AND METHODS 110
2.1 MATERIALS 110
2.2 METHODS 111
2.2.1 PLGA MODIFICATION 111
2.2.1.1 SYNTHESIS OF PEG-SPACER 111
2.2.1.2 SYNTHESIS OF PEG-PLGA 112
2.2.1.3 TNBS ASSAY 113
2.2.2 COATING PROCEDURE FOR PEG-PLGA 114
2.2.3 VEGFI65 LINKAGE TO MODIFIED PLGA MATRIX 115
2.2.4 EVALUATION OF DETECTION METHODS FOR SURFACE BOUND VEGFRES 115
2.2.4.1 ATTENUATED TOTAL REFLECTION- FT-IR SPECTROSCOPY (ATR-FTIR) 115
2.2.4.2 VEGFWS DETECTION USING STAINING METHODS 116
2.2.4.3 ANTIBODY DETECTION 116
2.2.5 INFLUENCE ON CELL GROWTH BY COVALENT LINKAGE OF VEGF165 116
2.2.5.I VASCULAR ENDOTHELIAL CELLS 116
2.2.5.2 RAT AORTIC SMOOTH MUSCLE CELLS 117
2.2.5.3 CELL GROWTH STUDY. 117
3 RESULTS AND DISCUSSION 118
3.1 PLGA MODIFICATION 118
3.2 COVALENT LINKAGE OF VEGF 165 TO MODIFIED PLGA MATRICES 119
3.2.1 COMPARISON OF VEGFRES DETECTION METHODS 119
3.2.1.1 FT-IR SPECTROSCOPY 120
3.2.1.2 REVERSIBLE PROTEIN DETECTION KIT....: 122
3.2.1.3 SILVER STAINING OF COVALENTLY LINKED VEGF,65 123
3.2.1.4 DEEP PURPLE* TOTAL PROTEIN STAIN FOR THE DETECTION OFVEGFIES 124
3.2.1.5 ANTIBODY DETECTION OF COVALENTLY LINKED VEGF T65 125
3.2.2 INFLUENCE OF PLGA COATING CONCENTRATION ON VEGFIES LINKAGE 130
3.2.3 INFLUENCE OF VEGF, 65 CONCENTRATION ON THE LINKAGE PROCEDURE 131
3.3 INFLUENCE OF COVALENTLY LINKED VEGF165 ON CELL GROWTH 132
3.3.1 VASCULAR ENDOTHELIAL CELLS 132
3.3.2 CELL GROWTH COMPARISON STUDY FOR SMOOTH MUSCLE AND ENDOTHELIAL
CELLS 136
4 CONCLUSIONS 138
5 REFERENCES 139
IMAGE 6
TABLE OF CONTENTS
CHAPTER 6
CELL GROWTH STIMULATION BY COVALENT LINKAGE OF VEGF 16S TO A MODIFIED
COLLAGEN-MATRIX 143
1 INTRODUCTION 144
2 MATERIALS AND METHODS 145
2.1 MATERIALS 145
2.2 METHODS 146
2.2.1 SYNTHESIS OF PEG-SPACER 146
2.2.2 TNBS ASSAY 147
2.2.3 COLLAGEN COATING PROCEDURE 148
2.2.3.1 PREPARATION OF COLLAGEN DISPERSION 148
2.2.3.2 COATING PROCEDURE 148
2.2.4 VEGFRES LINKAGE 149
2.2.4.1 PEG-SPACER COUPLING 149
2.2.4.2 VEGFMS LINKAGE PROCEDURE 150
2.2.5 ANTIBODY DETECTION OF SURFACE BOUND VEGFI65 150
2.2.6 INFLUENCE OF COVALENTLY LINKED VEGF165 ON CELL GROWTH 151
2.2.6.1 VASCULAR ENDOTHELIAL CELLS 151
2.2.6.2 AORTIC SMOOTH MUSCLE CELLS 151
2.2.6.3 CELL GROWTH COMPARISON STUDY 151
3 RESULTS AND DISCUSSION 152
3.1 COVALENT VEGF165 LINKAGE TO MODIFIED COLLAGEN MATRICES 152
3.1.1 EVALUATION OF ANTIBODY STAINING METHOD 152
3.1.2 ANALYSIS OF THE LINKAGE PROCEDURE OF VEGF TO PEG-COLLAGEN 153
3.2 INFLUENCE OF VEGF 165 COVALENTLY LINKED TO PEG-COLLAGEN ON CELL
GROWTH 157
3.2.1 VASCULAR ENDOTHELIAL CELLS 157
3.2.2 CELL GROWTH COMPARISON STUDY 159
4 CONCLUSIONS 163
5 REFERENCES 164
CHAPTER 7
SUMMARY OF THE THESIS 167
|
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| ctrlnum | (OCoLC)706987357 (DE-599)DNB100933039X |
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| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 612 - Human physiology |
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| dewey-sort | 3612.13 |
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| discipline | Medizin |
| edition | 1. Aufl. |
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| institution | BVB |
| isbn | 9783868537567 |
| language | English |
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| physical | 169 S. Ill., graph. Darst. |
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| spelling | Schaubhut, Frank 1979- Verfasser (DE-588)143342371 aut VEGF carrying coatings for enhanced vascular graft endothelialization Frank Schaubhut 1. Aufl. München Verl. Dr. Hut 2010 169 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Pharmazeutische Technologie Zugl.: München, Univ., Diss., 2010 Angiogenese (DE-588)4142449-9 gnd rswk-swf Vascular endothelial Growth Factor (DE-588)4319404-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Angiogenese (DE-588)4142449-9 s Vascular endothelial Growth Factor (DE-588)4319404-7 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-125453 http://edoc.ub.uni-muenchen.de/12545/ Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-125453 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=021097575&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schaubhut, Frank 1979- VEGF carrying coatings for enhanced vascular graft endothelialization Angiogenese (DE-588)4142449-9 gnd Vascular endothelial Growth Factor (DE-588)4319404-7 gnd |
| subject_GND | (DE-588)4142449-9 (DE-588)4319404-7 (DE-588)4113937-9 |
| title | VEGF carrying coatings for enhanced vascular graft endothelialization |
| title_auth | VEGF carrying coatings for enhanced vascular graft endothelialization |
| title_exact_search | VEGF carrying coatings for enhanced vascular graft endothelialization |
| title_full | VEGF carrying coatings for enhanced vascular graft endothelialization Frank Schaubhut |
| title_fullStr | VEGF carrying coatings for enhanced vascular graft endothelialization Frank Schaubhut |
| title_full_unstemmed | VEGF carrying coatings for enhanced vascular graft endothelialization Frank Schaubhut |
| title_short | VEGF carrying coatings for enhanced vascular graft endothelialization |
| title_sort | vegf carrying coatings for enhanced vascular graft endothelialization |
| topic | Angiogenese (DE-588)4142449-9 gnd Vascular endothelial Growth Factor (DE-588)4319404-7 gnd |
| topic_facet | Angiogenese Vascular endothelial Growth Factor Hochschulschrift |
| url | http://edoc.ub.uni-muenchen.de/12545/ https://nbn-resolving.org/urn:nbn:de:bvb:19-125453 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=021097575&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT schaubhutfrank vegfcarryingcoatingsforenhancedvasculargraftendothelialization |