Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
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2016
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| 007 | t| | ||
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| 082 | 0 | |a 570 |2 23 | |
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| 084 | |a 570 |2 sdnb | ||
| 100 | 1 | |a Rossi, Angela Francesca |d 1984- |e Verfasser |0 (DE-588)1138592315 |4 aut | |
| 245 | 1 | 0 | |a Development of functionalized electrospun fibers as biomimetic artificial basement membranes |b = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen |c submitted by Angela Francesca Rossi from Ulml |
| 246 | 1 | 1 | |a Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen |
| 264 | 1 | |a Würzburg |c 2016 | |
| 300 | |a XXI, 198 Blätter, Appendix: 35 ungezählte Blätter |b Illustrationen, Diagramme | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a veröffentlicht 2017 | ||
| 502 | |b Dissertation |c Julius-Maximilians-Universität Würzburg |d 2016 | ||
| 538 | |a Langzeitarchivierung gewährleistet, LZA | ||
| 546 | |a Zusammenfassung in deutscher und englischer Sprache | ||
| 650 | 0 | 7 | |a Basalmembran |0 (DE-588)4144086-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Tissue Engineering |0 (DE-588)4646061-5 |2 gnd |9 rswk-swf |
| 653 | |a Skin | ||
| 653 | |a Tissue Engineering | ||
| 653 | |a Bipolar | ||
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| 912 | |a ebook | ||
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-029855995 | |
Datensatz im Suchindex
| DE-BY-TUM_katkey | 2279515 |
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| _version_ | 1820892410770620416 |
| adam_text | TABLE OF CONTENTS
LIST OF
FIGURES........................................................................................................................
VII
LIST OF
TABLES............................................................................................................................
X
ABBREVIATIONS.........................................................................................................................XI
SUMMARY..............................................................................................................................
XVI
ZUSAMMENFASSUNG.............................................................................................................
XIX
1
INTRODUCTION.....................................................................................................................
1
1.1 TISSUE ENGINEERING - PRINCIPLES AND
APPLICATIONS..............................................1
1.2 SKIN AND ORAL MUCOSA TISSUE
ENGINEERING............................................................4
1.2.1 ANATOMY OF THE SKIN AND THE ORAL M UCOSA
..................................................
4
1.2.2 SKIN
EQUIVALENTS..............................................................................................8
1.2.3 ORAL MUCOSA
EQUIVALENTS.............................................................................
10
1.3 THE ROLE OF THE BASEMENT MEMBRANE IN THE SKIN AND ORAL M UCOSA
...............
12
1.3.1 BASEMENT M EM BRANE COM POSITION
...........................................................
13
1.3.2 BASEMENT M EM BRANE ASSEM
BLY.................................................................14
1.3.3 BASEMENT M EM BRANE FUNCTION
..................................................................
15
1.4 DEVELOPMENT OF ARTIFICIAL BASEMENT MEMBRANES
.............................................
16
1.4.1 E
LECTROSPINNING............................................................................................
17
1.4.2 MODIFICATION OF ELECTROSPUN M EM B RAN
ES.................................................22
1.5 SCOPE AND STRUCTURE OF THE THESIS
........
.
.............................................................
29
2
MATERIALS........................................................................................................................32
2.1 CELL
SOURCES............................................................................................................32
2.1.1 CELL LIN E
..........................................................................................................
32
2.1.2 PRIM ARY SKIN
CELLS.........................................................................................32
2.1.3 PRIM ARY ORAL MUCOSA CELLS
..........................................................................
32
2.2 EQUIPMENT
2.3 DISPOSABLE MATERIAL
.............................
35
2.4 LABORATORY
MATERIAL.............................................................................................
36
2.5 CHEMICALS AND
SOLUTIONS......................................................................................
37
2.6 GENERAL BUFFERS AND WORKING
SOLUTIONS...............................................................40
2.7 BUFFERS AND SOLUTIONS FOR CELL
CULTURE.................................................................41
2.8 MEDIA FOR CELL CULTURE
...........................................................................................
42
2.9 CHEMICALS AND SOLUTIONS FOR HISTOLOGY AND IMMUNOHISTOCHEMISTRY.
............
43
2.10 KITS FOR
IMMUNOHISTOCHEMISTRY......................................................................
44
2.11 ANTIBODIES FOR
IMMUNOHISTOCHEMISTRY..........................................................45
2.12 PEPTIDE SEQUENCES FOR
ELECTROSPINNING..........................................................45
2.13
SOFTWARE............................................................................
46
3
METHODS.........................................................................................................................
47
3.1 PREPARATION OF MEMBRANES FOR TISSUE M
ODELS..................................................47
3.1.1 COLLAGEN TYPE I
HYDROGEL.............................................................................
47
3.1.2 SOLUTION FOR
ELECTROSPINNING.......................................................................48
3.1.3 ELECTROSPINNING OF PLGA M EM
BRANES.......................................................48
3.2 CELL CULTURE
METHODS............................................................................................
48
3.2.1 ISOLATION OF PRIM ARY CELLS FROM HUM AN SKIN B IO P SIES
............................
49
3.2.2 ISOLATION OF PRIM ARY CELLS FROM HUM AN ORAL MUCOSA BIOPSIES
..............
50
3.2.3 CULTURE CONDITIONS OF CELLS
..........................................................................
51
3.2.4 PASSAGING OF
CELLS.........................................................................................
52
3.2.5 D ETERM INATION OF CELL NUM BER AND VIABILITY
............................................
52
3.2.6 FREEZING AND THAW ING OF
CELLS....................................................................
53
3.2.7 CELL CULTURE ON CHAM BER SLIDES
54
3.2.8 CELL CULTURE ON
CYTOSPOTS.............................................................................
54
3.2.9 FIXATION OF MEM BRANES FOR STATIC CELL CU LTU
RE..........................................54
3.2.10 DYNAMIC CULTURE OF MEM BRANES IN FLOW -BIOREACTOR
...............................
55
3.3 CONSTRUCTION OF TISSUE MODELS
...........................................................................
55
3.3.1 M ONO-CULTURE OF CELLS ON FUNCTIONALIZED M EM B RAN ES
............................
56
3.3.2 CO-CULTURE OF CELLS ON FUNCTIONALIZED M EM BRANES
..................................
56
3.3.3 M ONO-CULTURE OF CELLS ON POLYCARBONATE M EM BRANES
............................
56
3.3.4 CO-CULTURE OF CELLS ON COLLAGEN I
HYDROGELS..............................................57
3.4 CHARACTERIZATION OF FUNCTIONALIZED MEMBRANES AND TISSUE MODELS
.............
57
3.4.1 SCANNING ELECTRON M ICROSCOPY
...................................................................
57
3.4.2 QUALITATIVE PROTEIN ADSORPTION STU D IE S
....................................................
58
3.4.3 QUANTIFICATION OF PEPTIDE SEQUENCES........................ 58
3.4.4 LIVE/DEAD
STAINING.......................................................................................59
3.4.5 INVASIVE CELL VIABILITY ASSAY (M
TT-ASSAY)................................................60
3.4.6 NON-INVASIVE CELL VIABILITY ASSAY (CELLTITER-FLUOR* )
............................
61
3.4.7 TRANSEPITHELIAL ELECTRICAL RESISTAN CE
........................................................
61
3.4.6 PERM EATION STU D
IES.......................................................................................61
3.4.9 ET-50 M EASUREM
ENT....................................................................................
63
3.4.10 IRRITATION STU D Y
.............................................................................................63
3.5
HISTOLOGY...............................................................................................................
64
3.5.1
FIXATION...................................................................................
64
3.5.2 PLUNGE FREEZING AND CRYO-SECTIONING
........................................................
64
3.5.3 PARAFFIN EM BEDDING AND M ICROTOME
SECTIONING......................................65
3.5.4 DEPARAFFINIZATION AND REHYDRATION OF TISSUE SECTIO N S
...........................
66
3.5.5 HAEMATOXYLIN-EOSIN STAINING
.67
3.5.6 IM M UNOHISTOCHEMISTRY,
3.6 DEGRADATION STUDIES
.............................
68
3.7 DIFFERENTIAL SCANNING CALORIMETRY
......................................................................
68
3.8
LYOPHILIZATION........................................................................................................
69
3.9 STATISTICAL
ANALYSIS.................................................................................................69
4
RESULTS............................................................................................................................70
4.1 CHARACTERIZATION OF FUNCTIONALIZED MEMBRANES
...............................................
70
4.1.1 ELECTROSPINNING OF FUNCTIONALIZED M EM BRANES
.......................................
70
4.1.2 INFLUENCE OF PEPTIDES ON FIBER MORPHOLOGY AND INTEGRITY
.....................
71
4.1.3 INFLUENCE OF FIXATION M ODE ON FIBER M ORPHOLOGY
....................................
73
4.1.4 MODIFYING PORE SIZE OF FUNCTIONALIZED M EM B RAN
ES................................74
4.1.5 DEGRADATION OF FUNCTIONALIZED M EM B RAN ES
............
.
................................
77
4.1.6 INFLUENCE OF STERILIZATION ON M EM BRANE M ORPHOLOGY
............................
79
4.1.7 CULTURE OF FUNCTIONALIZED M EM BRANES U NDER DYNAMIC CONDITIONS
.....
81
4.1.8 SUM M
ARY........................................................................................................87
4.2 QUANTIFICATION OF IMMOBILIZED PEPTIDES ON FUNCTIONALIZED MEMBRANES
......
88
4.2.1 ELECTROSPINNING OF MEM BRANES W ITH QUANTIFIABLE PEPTIDES
.................
88
4.2.2 QUANTIFICATION OF PEPTIDE CONCENTRATION
..................................................
91
4.2.3 QUANTIFICATION OF PEPTIDE BINDING M ODE
..................................................
93
4.2.4 COMPARISON OF EXPECTED AND M EASURED PEPTIDE CONCENTRATION
..........
95
4.2.5 INFLUENCE OF PEPTIDE CONCENTRATION ON
BINDING.......................................96
4.2.6 INFLUENCE OF INCUBATION ON PEPTIDE B IN D IN G
............................................
97
4.2.7 SUM M
ARY........................................................................................................98
4.3 BIOLOGICAL INTERACTIONS OF FUNCTIONALIZED MEMBRANES IN MONO-CULTURE
.......
99
4.3.1 PROTEIN ADSORPTION ON M EM BRANES
............................................................
99
4.3.2 FIBROBLAST ADHESION AND PROLIFERATION ON M EM B RAN ES
.......................
101
4.3.3 HACAT ADHESION AND PROLIFERATION ON M E M B RA N ES
.............................
104
4.3.4 SUM M
ARY......................................................................................................107
4.4 FUNCTIONALIZED MEMBRANES FOR SKIN TISSUE
ENGINEERING..............................108
4.4.1 ISOTROPIC M EM BRANES FOR SKIN TISSUE ENGINEERING
...............................
108
4.4.2 BIPOLAR M EM BRANES FOR SKIN TISSUE ENGINEERING
..................................
113
4.4.3 SUM M
ARY......................................................................................................116
4.5 FUNCTIONALIZED MEMBRANES FOR PRIMARY SKIN
CELLS.......................................117
4.5.1 FUNCTIONALIZED M EM BRANES FOR EPITHELIAL SKIN M ODELS
.......................
117
4.5.2 FUNCTIONALIZED M EM BRANES FOR CO-CULTURE SKIN M
ODELS......................124
4.5.3 INFLUENCE OF PORE SIZE ON KERATINOCYTE D EV ELO P M EN T
..........................
126
4.5.4 INFLUENCE OF PH VALUE ON KERATINOCYTE VIABILITY
....
.
..............................127
4.5.5 SUM M
ARY......................................................................................................128
4.6 FUNCTIONALIZED MEMBRANES FOR ORAL MUCOSA TISSUE
ENGINEERING................129
4.6.1 ISOLATION OF ORAL MUCOSA
CELLS...................................................................129
4.6.2 CHARACTERIZATION OF ORAL MUCOSA
CELLS.....................................................130
4.6.3 FUNCTIONALIZED M EM BRANES FOR ORAL MUCOSA M
ODELS...........................130
4.6.4 POLYCARBONATE M EM BRANES FOR ORAL MUCOSA M
ODELS...........................131
4.6.5 SUM M
ARY......................................................................................................140
5
DISCUSSION..................................................................................................................
141
5.1 CHARACTERIZATION OF FUNCTIONALIZED MEMBRANES
.......................
.
...................
141
5.2 QUANTIFICATION OF IMMOBILIZED PEPTIDES ON FUNCTIONALIZED
MEMBRANES.... 147
5.3 BIOLOGICAL INTERACTIONS OF FUNCTIONALIZED MEMBRANES IN
MONO-CULTURE..... 153
5.3.1 PROTEIN ADSORPTION ON FUNCTIONALIZED M EM BRANES
...............................
154
5.3.2 CELL ADHESION AND PROLIFERATION ON FUNCTIONALIZED M EM B RAN ES
.......
156
5.4 FUNCTIONALIZED MEMBRANES FOR SKIN TISSUE ENGINEERING
5.4.1 ISOTROPIC M EM BRANES FOR SKIN TISSUE ENGINEERING
..............................
160
5.4.2 BIPOLAR M EM BRANES FOR SKIN TISSUE ENGINEERING
.................................
161
5.5 FUNCTIONALIZED MEMBRANES FOR PRIMARY SKIN CELLS
.......................................
165
5.5.1 FUNCTIONALIZED MEM BRANES FOR EPITHELIAL SKIN M ODELS
........................
166
5.5.2 FUNCTIONALIZED MEM BRANES FOR CO-CULTURE SKIN M ODELS
......................
167
5.6 FUNCTIONALIZED MEMBRANES FOR ORAL MUCOSA TISSUE ENGINEERING
................
172
5.7 FUTURE
PERSPECTIVES............................................................................................
178
6
REFERENCES...................................................................................................................
181
7
APPENDIX......................................................................................................................199
7.1 STANDARD OPERATING PROCEDURE
PROTOCOLS.......................................................199
7.2 PUBLICATIONS AND CONFERENCE
CONTRIBUTIONS....................................................226
7.3 CURRICULUM
VITAE.................................................................................................
229
7.4
ACKNOWLEDGMENT...............................................................................................
230
7.5
AFFIDAVIT...............................................................................................................
232
7.6 EIDESSTATTLICHE ERKLAERUNG, 233
|
| any_adam_object | 1 |
| author | Rossi, Angela Francesca 1984- |
| author_GND | (DE-588)1138592315 |
| author_facet | Rossi, Angela Francesca 1984- |
| author_role | aut |
| author_sort | Rossi, Angela Francesca 1984- |
| author_variant | a f r af afr |
| building | Verbundindex |
| bvnumber | BV044455187 |
| collection | ebook |
| ctrlnum | (OCoLC)1159313872 (DE-599)DNB1137835915 |
| dewey-full | 570 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 570 - Biology |
| dewey-raw | 570 |
| dewey-search | 570 |
| dewey-sort | 3570 |
| dewey-tens | 570 - Biology |
| discipline | Biologie Medizin |
| format | Thesis Book |
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| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-029855995 |
| oclc_num | 1159313872 |
| open_access_boolean | 1 |
| owner | DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-12 DE-91 DE-BY-TUM DE-19 DE-BY-UBM DE-1049 DE-92 DE-739 DE-898 DE-BY-UBR DE-355 DE-BY-UBR DE-706 DE-20 DE-1102 DE-860 DE-2174 |
| owner_facet | DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-12 DE-91 DE-BY-TUM DE-19 DE-BY-UBM DE-1049 DE-92 DE-739 DE-898 DE-BY-UBR DE-355 DE-BY-UBR DE-706 DE-20 DE-1102 DE-860 DE-2174 |
| physical | XXI, 198 Blätter, Appendix: 35 ungezählte Blätter Illustrationen, Diagramme |
| psigel | ebook |
| publishDate | 2016 |
| publishDateSearch | 2016 |
| publishDateSort | 2016 |
| record_format | marc |
| spellingShingle | Rossi, Angela Francesca 1984- Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen Basalmembran (DE-588)4144086-9 gnd Tissue Engineering (DE-588)4646061-5 gnd |
| subject_GND | (DE-588)4144086-9 (DE-588)4646061-5 (DE-588)4113937-9 |
| title | Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen |
| title_alt | Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen |
| title_auth | Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen |
| title_exact_search | Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen |
| title_full | Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen submitted by Angela Francesca Rossi from Ulml |
| title_fullStr | Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen submitted by Angela Francesca Rossi from Ulml |
| title_full_unstemmed | Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen submitted by Angela Francesca Rossi from Ulml |
| title_short | Development of functionalized electrospun fibers as biomimetic artificial basement membranes |
| title_sort | development of functionalized electrospun fibers as biomimetic artificial basement membranes entwicklung funktionalisierter elektrogesponnener fasern als biomimetische kunstliche basalmembranen |
| title_sub | = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen |
| topic | Basalmembran (DE-588)4144086-9 gnd Tissue Engineering (DE-588)4646061-5 gnd |
| topic_facet | Basalmembran Tissue Engineering Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-137618 http://d-nb.info/1137835915/34 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029855995&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT rossiangelafrancesca developmentoffunctionalizedelectrospunfibersasbiomimeticartificialbasementmembranesentwicklungfunktionalisierterelektrogesponnenerfasernalsbiomimetischekunstlichebasalmembranen AT rossiangelafrancesca entwicklungfunktionalisierterelektrogesponnenerfasernalsbiomimetischekunstlichebasalmembranen |