Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2008
|
| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:29-opus-10596 http://d-nb.info/990011305/34 Inhaltsverzeichnis |
| Beschreibung: | 148 Bl. Ill., graph. Darst. |
Internformat
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Datensatz im Suchindex
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|---|---|
| adam_text | Inhaltsverzeichnis
Inhaltsverzeichnis/lndex
Inhaltsverzeichnis/lndex 5
Einleitung (deutsch) 10
Abstract 11
Abbreviations 12
1. Introduction 17
1.1. The cornea 17
1.1.1. Corneal epithelium 17
1.1.2. Bowman s layer 18
1.1.3. Corneal stroma 18
1.1.4. Descement s membrane 19
1.1.5. Corneal endothelium 19
1.1.6. Corneal innervations 21
1.1.7. Corneal Limbus 22
1.1.8. Trabecular Meshwork 23
1.2. Angiogenic privilege of the cornea 24
1.3. Endogenous inhibitors of angiogenesis in the cornea and aqueous
humor 27
1.3.1. Angiostatin 27
1.3.2. Endostatin 28
1.3.3. Pigment Epithelium-Derived Factor 28
1.3.4. Thrombospondin 29
1.4. Immune privilege of the eye 30
1.5. The role of angiogenic and immune privilege of the cornea in corneal
transplant immunology 33
1.6. Angiogenesis and lymphangiogenesis in the cornea 35
1.7. Clinical consequences of loss of corneal angiogenic privilege 40
1.8. Exogenous inhibitors of angiogenesis 43
1.8.1. Bevacizumab 44
5
Inhaltsverzeichnis
1.8.2. VEGFR3 antibody mF4-31C1 45
1.8.3. VEGFTrapRiR2 45
1.9. Morphometrical methods to quantify corneal hem- and
lymphangiogenesis 46
2. Material and Methods 49
2.1. Material 49
2.1.1. Chemicals and expendables 49
2.1.2. Surgical instruments 50
2.1.3. Primary antibodies 51
2.1.4. Secondary antibodys 51
2.1.5. Eukaryotic cells 52
2.1.6. Animals 52
2.1.7. Software 52
2.3. Methods 53
2.3.1. Cell culture 53
2.3.2. Lymphatic endothelial cell (LEC)/ blood endothelial cell (BEC)
proliferation ELISA 54
2.3.3. Bevacizumab binding analysis with murine and human VEGF-A 55
2.3.3.1. Western blot 55
2.3.3.2. Solid phase binding assay (ELISA) 56
2.3.3.3. Surface plasmon resonance (BIACORE®) 57
2.3.4. Animal models 58
2.3.4.1. Aqueous humor accumulation 58
2.3.4.2. Wound healing assay 58
2.3.4.3. Alkali Burn assay 59
2.3.4.4. Suture-induced, inflammatory corneal neovascularization assay 59
2.3.4.5. Murine corneal transplantation 60
2.3.5. Immunohistochemical methods 61
2.3.5.1. Corneal wholemounts 61
2.3.5.2. Cryosection staining 62
2.3.5.3. Light Microscopy 62
2.3.6. Functional analysis 63
2.3.6.1. Image acquisition 63
6
Inhaltsverzeichnis
2.3.6.2. Manual method of cornea! image analysis using ImageJ 64
2.3.6.3. Semiautomatic method of corneal image analysis using CellAF 64
2.3.6.4. Statistical analysis 68
3. Results 69
3.1. Development of a new semiautomatic, quantitative method for
morphological analysis of hem- and lymphangiogenesis in corneal
flatmounts 69
3.1.1. Validity of manual outlining versus semiautomatic threshold
method analyzing model structures 69
3.1.2. Comparison of the validity of the manual versus the semiautomatic
method analyzing blood and lymphatic vessels in corneal
flatmounts 71
3.1.3. Comparison of the reproducibility and objectivity of the two
methods 75
3.2. Effects of the first FDA-approved antiangiogenic drug Bevacizumab
on corneal hem- and lymphangiogenesis 77
3.2.1. Bevacizumab binds to murine and human VEGF-A 78
3.2.2. Systemic application of Bevacizumab inhibits hem- and
lymphangiogenesis 82
3.2.3. Topical application of Bevacizumab inhibits hem- and
lymphangiogenesis 84
3.2.4. Long term application of Bevacizumab 85
3.2.5. IgG-isotype control backup 85
3.2.6. Bevacizumab inhibits lymphatic endothelial cell (LEC) proliferation
in vitro 86
3.2.7. Safety profile of topical VEGF-neutralization at the cornea using
Bevacizumab eye drops 88
3.2.7.5. Effects of Bevacizumab eye drops on normal murine corneas in
vivo 88
3.2.7.6. Effects of Bevacizumab eye drops on corneal epithelial wound
healing 91
3.3. Blockade of VEGFR3-signalling specifically inhibits
lymphangiogenesis in inflammatory corneal neovascularization 95
7
Inhaltsverzeichnis
3.3.1. Systemic application of mF4-31C1 inhibits lymph-but not
hemangiogenesis 95
3.3.2. The role of lymphatic vessels for cell debris clearing during wound
healing in the cornea 99
3.3.3. Improvement of graft survival after high risk keratoplasty by
blocking blood and lymphatic vessels or only lymphatic vessels 100
3.3.3.1. Pretreatment with the VEGF TrapR1R2 101
3.3.3.2. Pretreatment with the VEGFR3 antibody mF4-31 C1 102
3.4. Immune modulatory factors in the aqueos humor have
antilymphangiogenic effects 103
3.4.1. Antihem- and antilymphangiogenic effects of the aqueous humor 104
3.4.2. The endogenous, antiangiogenic factor thrombospondin 1 has
antilymphangiogenic effects in vitro 104
3.4.3. The endogenous immune modulating factors a-melanocyte
stimulating hormone (a -MSH) and vasoactive intestinal protein
(VIP) exhibit various effects on hem- and lymphangiogenesis in
vitro 105
3.4.3.3. a-MSH 105
3.4.3.4. VIP 106
4. Discussion 111
4.1. Improved semiautomatic method for morphometry of angiogenesis
and lymphangiogenesis in corneal flatmounts 113
4.2. Bevacizumab is the first FDA-approved antiangiogenic drug, which
also inhibits lymphangiogenesis in addition to hemangiogenesis 114
4.3. Bevacizumab eye drops do not seem to affect normal corneal
structure or epithelial wound healing 117
4.4. The role of lymphatic vessels in inflammatory corneal
neovascularization 120
4.5. Aqueous humor is a potent model to identify novel
antilymphangiogenic factors 126
5. References 132
8
Inhaltsverzeichnis
6. Lebenslauf und Publikationsliste 141
Lebenslauf 141
Preise 144
Publikationsliste 144
Danksagung Fehler! Textmarke nicht definiert.
9
|
| adam_txt |
Inhaltsverzeichnis
Inhaltsverzeichnis/lndex
Inhaltsverzeichnis/lndex 5
Einleitung (deutsch) 10
Abstract 11
Abbreviations 12
1. Introduction 17
1.1. The cornea 17
1.1.1. Corneal epithelium 17
1.1.2. Bowman's layer 18
1.1.3. Corneal stroma 18
1.1.4. Descement's membrane 19
1.1.5. Corneal endothelium 19
1.1.6. Corneal innervations 21
1.1.7. Corneal Limbus 22
1.1.8. Trabecular Meshwork 23
1.2. Angiogenic privilege of the cornea 24
1.3. Endogenous inhibitors of angiogenesis in the cornea and aqueous
humor 27
1.3.1. Angiostatin 27
1.3.2. Endostatin 28
1.3.3. Pigment Epithelium-Derived Factor 28
1.3.4. Thrombospondin 29
1.4. Immune privilege of the eye 30
1.5. The role of angiogenic and immune privilege of the cornea in corneal
transplant immunology 33
1.6. Angiogenesis and lymphangiogenesis in the cornea 35
1.7. Clinical consequences of loss of corneal angiogenic privilege 40
1.8. Exogenous inhibitors of angiogenesis 43
1.8.1. Bevacizumab 44
5
Inhaltsverzeichnis
1.8.2. VEGFR3 antibody mF4-31C1 45
1.8.3. VEGFTrapRiR2 45
1.9. Morphometrical methods to quantify corneal hem- and
lymphangiogenesis 46
2. Material and Methods 49
2.1. Material 49
2.1.1. Chemicals and expendables 49
2.1.2. Surgical instruments 50
2.1.3. Primary antibodies 51
2.1.4. Secondary antibodys 51
2.1.5. Eukaryotic cells 52
2.1.6. Animals 52
2.1.7. Software 52
2.3. Methods 53
2.3.1. Cell culture 53
2.3.2. Lymphatic endothelial cell (LEC)/ blood endothelial cell (BEC)
proliferation ELISA 54
2.3.3. Bevacizumab binding analysis with murine and human VEGF-A 55
2.3.3.1. Western blot 55
2.3.3.2. Solid phase binding assay (ELISA) 56
2.3.3.3. Surface plasmon resonance (BIACORE®) 57
2.3.4. Animal models 58
2.3.4.1. Aqueous humor accumulation 58
2.3.4.2. Wound healing assay 58
2.3.4.3. Alkali Burn assay 59
2.3.4.4. Suture-induced, inflammatory corneal neovascularization assay 59
2.3.4.5. Murine corneal transplantation 60
2.3.5. Immunohistochemical methods 61
2.3.5.1. Corneal wholemounts 61
2.3.5.2. Cryosection staining 62
2.3.5.3. Light Microscopy 62
2.3.6. Functional analysis 63
2.3.6.1. Image acquisition 63
6
Inhaltsverzeichnis
2.3.6.2. Manual method of cornea! image analysis using ImageJ 64
2.3.6.3. Semiautomatic method of corneal image analysis using CellAF 64
2.3.6.4. Statistical analysis 68
3. Results 69
3.1. Development of a new semiautomatic, quantitative method for
morphological analysis of hem- and lymphangiogenesis in corneal
flatmounts 69
3.1.1. Validity of manual outlining versus semiautomatic threshold
method analyzing model structures 69
3.1.2. Comparison of the validity of the manual versus the semiautomatic
method analyzing blood and lymphatic vessels in corneal
flatmounts 71
3.1.3. Comparison of the reproducibility and objectivity of the two
methods 75
3.2. Effects of the first FDA-approved antiangiogenic drug Bevacizumab
on corneal hem- and lymphangiogenesis 77
3.2.1. Bevacizumab binds to murine and human VEGF-A 78
3.2.2. Systemic application of Bevacizumab inhibits hem- and
lymphangiogenesis 82
3.2.3. Topical application of Bevacizumab inhibits hem- and
lymphangiogenesis 84
3.2.4. Long term application of Bevacizumab 85
3.2.5. IgG-isotype control backup 85
3.2.6. Bevacizumab inhibits lymphatic endothelial cell (LEC) proliferation
in vitro 86
3.2.7. Safety profile of topical VEGF-neutralization at the cornea using
Bevacizumab eye drops 88
3.2.7.5. Effects of Bevacizumab eye drops on normal murine corneas in
vivo 88
3.2.7.6. Effects of Bevacizumab eye drops on corneal epithelial wound
healing 91
3.3. Blockade of VEGFR3-signalling specifically inhibits
lymphangiogenesis in inflammatory corneal neovascularization 95
7
Inhaltsverzeichnis
3.3.1. Systemic application of mF4-31C1 inhibits lymph-but not
hemangiogenesis 95
3.3.2. The role of lymphatic vessels for cell debris clearing during wound
healing in the cornea 99
3.3.3. Improvement of graft survival after high risk keratoplasty by
blocking blood and lymphatic vessels or only lymphatic vessels 100
3.3.3.1. Pretreatment with the VEGF TrapR1R2 101
3.3.3.2. Pretreatment with the VEGFR3 antibody mF4-31 C1 102
3.4. Immune modulatory factors in the aqueos humor have
antilymphangiogenic effects 103
3.4.1. Antihem- and antilymphangiogenic effects of the aqueous humor 104
3.4.2. The endogenous, antiangiogenic factor thrombospondin 1 has
antilymphangiogenic effects in vitro 104
3.4.3. The endogenous immune modulating factors a-melanocyte
stimulating hormone (a -MSH) and vasoactive intestinal protein
(VIP) exhibit various effects on hem- and lymphangiogenesis in
vitro 105
3.4.3.3. a-MSH 105
3.4.3.4. VIP 106
4. Discussion 111
4.1. Improved semiautomatic method for morphometry of angiogenesis
and lymphangiogenesis in corneal flatmounts 113
4.2. Bevacizumab is the first FDA-approved antiangiogenic drug, which
also inhibits lymphangiogenesis in addition to hemangiogenesis 114
4.3. Bevacizumab eye drops do not seem to affect normal corneal
structure or epithelial wound healing 117
4.4. The role of lymphatic vessels in inflammatory corneal
neovascularization 120
4.5. Aqueous humor is a potent model to identify novel
antilymphangiogenic factors 126
5. References 132
8
Inhaltsverzeichnis
6. Lebenslauf und Publikationsliste 141
Lebenslauf 141
Preise 144
Publikationsliste 144
Danksagung Fehler! Textmarke nicht definiert.
9 |
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| 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-29T |
| physical | 148 Bl. Ill., graph. Darst. |
| psigel | ebook |
| publishDate | 2008 |
| publishDateSearch | 2008 |
| publishDateSort | 2008 |
| record_format | marc |
| spelling | Bock, Felix 1977- Verfasser (DE-588)136063047 aut Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea von Felix Bock 2008 148 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Erlangen-Nürnberg, Univ., Diss., 2008 Langzeitarchivierung Nationalbibliothek gewährleistet Hornhaut (DE-588)4160665-6 gnd rswk-swf Angiogenese (DE-588)4142449-9 gnd rswk-swf Lymphgefäß (DE-588)4137306-6 gnd rswk-swf Inhibitor (DE-588)4027007-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Hornhaut (DE-588)4160665-6 s Lymphgefäß (DE-588)4137306-6 s Angiogenese (DE-588)4142449-9 s Inhibitor (DE-588)4027007-5 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:29-opus-10596 https://open.fau.de/handle/openfau/689 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:29-opus-10596 Resolvingsystem http://d-nb.info/990011305/34 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016663050&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Bock, Felix 1977- Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea Hornhaut (DE-588)4160665-6 gnd Angiogenese (DE-588)4142449-9 gnd Lymphgefäß (DE-588)4137306-6 gnd Inhibitor (DE-588)4027007-5 gnd |
| subject_GND | (DE-588)4160665-6 (DE-588)4142449-9 (DE-588)4137306-6 (DE-588)4027007-5 (DE-588)4113937-9 |
| title | Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea |
| title_auth | Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea |
| title_exact_search | Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea |
| title_exact_search_txtP | Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea |
| title_full | Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea von Felix Bock |
| title_fullStr | Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea von Felix Bock |
| title_full_unstemmed | Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea von Felix Bock |
| title_short | Endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea |
| title_sort | endogenous and exogenous inhibitors of inflammatory lymphangiogenesis in the cornea |
| topic | Hornhaut (DE-588)4160665-6 gnd Angiogenese (DE-588)4142449-9 gnd Lymphgefäß (DE-588)4137306-6 gnd Inhibitor (DE-588)4027007-5 gnd |
| topic_facet | Hornhaut Angiogenese Lymphgefäß Inhibitor Hochschulschrift |
| url | https://open.fau.de/handle/openfau/689 https://nbn-resolving.org/urn:nbn:de:bvb:29-opus-10596 http://d-nb.info/990011305/34 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016663050&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT bockfelix endogenousandexogenousinhibitorsofinflammatorylymphangiogenesisinthecornea |