Development of functionalized electrospun fibers as biomimetic artificial basement membranes: = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Würzburg
2016
|
| Subjects: | |
| Online Access: | Volltext http://d-nb.info/1137835915/34 Inhaltsverzeichnis |
| Item Description: | veröffentlicht 2017 |
| Physical Description: | XXI, 198 Blätter, Appendix: 35 ungezählte Blätter Illustrationen, Diagramme |
| Format: | Langzeitarchivierung gewährleistet, LZA |
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MARC
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| 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 Tissue Engineering |0 (DE-588)4646061-5 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Basalmembran |0 (DE-588)4144086-9 |2 gnd |9 rswk-swf |
| 653 | |a Skin | ||
| 653 | |a Tissue Engineering | ||
| 653 | |a Bipolar | ||
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| 689 | 0 | 1 | |a Tissue Engineering |0 (DE-588)4646061-5 |D s |
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| 912 | |a ebook | ||
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-029855995 | |
Record in the Search Index
| _version_ | 1823056934696124416 |
|---|---|
| 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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| language | English |
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| physical | XXI, 198 Blätter, Appendix: 35 ungezählte Blätter Illustrationen, Diagramme |
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| publishDateSort | 2016 |
| record_format | marc |
| spelling | Rossi, Angela Francesca 1984- Verfasser (DE-588)1138592315 aut 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 Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen Würzburg 2016 XXI, 198 Blätter, Appendix: 35 ungezählte Blätter Illustrationen, Diagramme txt rdacontent n rdamedia nc rdacarrier veröffentlicht 2017 Dissertation Julius-Maximilians-Universität Würzburg 2016 Langzeitarchivierung gewährleistet, LZA Zusammenfassung in deutscher und englischer Sprache Tissue Engineering (DE-588)4646061-5 gnd rswk-swf Basalmembran (DE-588)4144086-9 gnd rswk-swf Skin Tissue Engineering Bipolar (DE-588)4113937-9 Hochschulschrift gnd-content Basalmembran (DE-588)4144086-9 s Tissue Engineering (DE-588)4646061-5 s DE-604 Erscheint auch als Online-Ausgabe, PDF urn:nbn:de:bvb:20-opus-137618 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-137618 Resolving-System kostenfrei Volltext http://d-nb.info/1137835915/34 Langzeitarchivierung Nationalbibliothek DNB Datenaustausch application/pdf 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 Inhaltsverzeichnis |
| spellingShingle | Rossi, Angela Francesca 1984- Development of functionalized electrospun fibers as biomimetic artificial basement membranes = Entwicklung funktionalisierter elektrogesponnener Fasern als biomimetische künstliche Basalmembranen Tissue Engineering (DE-588)4646061-5 gnd Basalmembran (DE-588)4144086-9 gnd |
| subject_GND | (DE-588)4646061-5 (DE-588)4144086-9 (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 | Tissue Engineering (DE-588)4646061-5 gnd Basalmembran (DE-588)4144086-9 gnd |
| topic_facet | Tissue Engineering Basalmembran 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 |