Die Wundheilung im Zusammenhang mit der Aktivierung des Wnt-Signalweg und Type 1 / Type 2 Immunity wurde im Maus Modell und in-vitro an verschiedenen Zeitpunkten untersucht. Es hat sich herausgestellt, dass der Wnt-Signalweg während der Spätphase der Wundheilung aktiviert wird, also unter Type 2 Immunity. Diese Arbeit deutet darauf hin, dass die Wundheilung über eine Modellierung des Wnt-Signalweg beschleunigt oder verbessert werden kann, allerdings nur an spezifischen Zeitpunkten.:LIST OF ABBREVIATIONS........................................................................................ 5
INDEX OF FIGURES..................................................................................................7
INDEX OF TABLES.....................................................................................................8
SUMMARY.................................................................................................................9
1 INTRODUCTION..............................................................................................11
1.1 Cutaneous wound healing...............................................................................11
1.2 Phases of wound healing................................................................................11
1.2.01 Inflammation............................................................................................11
1.2.02 Proliferation..............................................................................................13
1.2.03 Remodeling..............................................................................................16
1.3 Wnt-signaling...................................................................................................17
1.3.01 Wnt-ligands biogenesis and receptors....................................................17
1.3.02 Canonical and non canonical Wnt signaling...........................................20
1.3.03 Canonical Wnt/ß catenin signaling..........................................................20
1.4 Wound healing and canonical Wnt/ß catenin signaling...................................21
2 AIMS OF THE STUDY......................................................................................25
3 MATERIALS AND EXPERIMENTAL PROCEDURES........................................27
3.1 MATERIALS.....................................................................................................27
3.1.01 Instruments..............................................................................................27
3.1.02 Software..................................................................................................27
3.1.03 Websites..................................................................................................28
3.1.04 Consumables...........................................................................................28
3.1.05 Media and Buffers...................................................................................29
3.1.06 Reagents..................................................................................................29
3.1.07 Stimulation reagents................................................................................30
3.1.08 Anesthesia and analgesia used for the full-thickness skin wound model.......................................................................................................................30
3.1.09 Kits...........................................................................................................30
3.1.10 Quantitative reverse-transcription PCR primers......................................31
3.2 EXPERIMENTAL PROCEDURES.....................................................................32
3.2.01 Mice.........................................................................................................32
3.2.02 Cell Lines.................................................................................................32
3.2.03 Cell counting with trypan blue stain and hemocytometer method.........32
3.2.04 Preparation of murine H133WT immortalized primary keratinocytes cultures....................................................................................................................33
3.2.05 Preparation of RAW264.7 murine macrophages cultures.......................33
3.2.06 Preparation of primary murine fibroblasts cultures.................................33
3.2.07 Type 1 and type 2 immunity cell stimulation...........................................34
3.2.08 Cell RNA isolation....................................................................................34
3.2.09 Mice wounding........................................................................................35
3.2.10 Whole skin tissue isolation......................................................................36
3.2.11 Epidermal and dermal tissue isolation....................................................36
3.2.12 Tissue RNA isolation...............................................................................36
3.2.13 cDNA synthesis.......................................................................................36
3.2.14 Quantitative Real-Time PCR (qPCR).......................................................37
3.2.15 Statistical analysis...................................................................................37
4 EXPERIMENTAL RESULTS..............................................................................39
4.1 Canonical Wnt/ß-catenin signaling activity in different skin cell populations under type 1 and type 2 immunity cytokines stimulation in vitro............................39
4.2 Canonical Wnt/ß-catenin signaling activity under type 1 and type 2 immunity conditions during wound healing in vivo.................................................................42
4.3 Wnt-ligands expression in different skin cell populations under type 1 and type 2 immunity cytokines stimulation in vitro.........................................................44
4.4 Wnt ligands expression in epidermal and dermal fraction under type 1 and type 2 immunity conditions in vivo..........................................................................51
5 DISCUSSION...................................................................................................53
6 CONCLUSIONS...............................................................................................59
7 PERSPECTIVES...............................................................................................61
8 REFERENCES..................................................................................................63
9 APPENDIX.......................................................................................................71
9.1 Supplementary results.....................................................................................71
9.2 Erklärung über die eigenständige Abfassung der Arbeit.................................73
9.3 Curriculum Vitae..............................................................................................75
9.4 Acknowledgments...........................................................................................79
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:33844 |
| Date | 29 April 2019 |
| Creators | Ordieres Ruiz, Michelle Denise |
| Contributors | Universität Leipzig |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:doctoralThesis, info:eu-repo/semantics/doctoralThesis, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
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