Angiogenesis from a new perspective

................................................................................................................... 7 LIST OF PAPERS ........................................................................................................... 9 ABBREVIATIONS ....................................................................................................... 10 INTRODUCTION AND BACKGROUND .................................................................. 11 Ocular angiogenesis ......................................................................................... 12 The vascular system .......................................................................................... 13 Vasculogenesis ................................................................................................. 13 Angiogenesis ..................................................................................................... 14 Sprouting angiogenesis ................................................................................ 15 Intussusceptive angiogenesis ....................................................................... 16 Looping angiogenesis .................................................................................. 17 Angiogenic switch ............................................................................................. 17 Inflammation and angiogenesis ........................................................................ 18 Tumor angiogenesis ......................................................................................... 19 Lymphatic vascular system ............................................................................... 19 The cornea ........................................................................................................ 20 Corneal anatomy .............................................................................................. 21 Ocular immune privilege .................................................................................. 22 Corneal avascularity and angiogenesis ........................................................... 22 Corneal lymphangiogenesis ............................................................................. 23 Treatments for angiogenesis ............................................................................ 26 Imaging of the cornea in the clinics ................................................................. 27 Confocal microscopy ........................................................................................ 28 In vivo confocal microscopy in the clinics ....................................................... 31 Imaging modalities for angiogenesis ............................................................... 31 Examples of current in vitro and in vivo angiogenesis assays ......................... 32 In vitro assays .............................................................................................. 32 In vivo assays ............................................................................................... 33 Corneal angiogenesis assay ............................................................................. 34 The rat corneal suture in vivo imaging assay .................................................. 35 General aim: ..................................................................................................... 37 Specific aims: ............................................................................................... 37 MATERIALS & METHODS ........................................................................................ 39 Experimental rat model of corneal neovascularization (I, III, IV) .................. 39 Initiation of capillary regression (IV) .............................................................. 39 Patient characteristics (II, IV) .......................................................................... 40 In vivo confocal microscopy (I-IV) ................................................................... 40 Cells within lymph and blood vessels (I, II) and measurement of lumen contrast ratio (I) ............................................................................................... 41 Quantification of vessel lumen diameter (I-IV) ................................................ 42 In vivo analysis of inflammatory cell infiltration (III, IV)................................ 42 Grading of neovascularization of rat corneas from photos (IV) ...................... 42 Identifying cell types (I, III, IV) ........................................................................ 42 Immunohistochemistry (I, III, IV) ..................................................................... 43 In vivo and ex vivo correlation of corneal structures (I, III, IV)...................... 45 Statistical analysis (I-IV) .................................................................................. 45 RESULTS ...................................................................................................................... 47 Study I .............................................................................................................. 47 Study II .............................................................................................................. 48 Study III ............................................................................................................ 49 Study IV ............................................................................................................ 50 DISCUSSION ................................................................................................................ 55 In vivo imaging of lymph vessels (I, II) ............................................................ 56 Hemangiogenesis (III, IV) ................................................................................ 57 Sprouting angiogenesis (III) ........................................................................ 57 Capillary regression (IV) ............................................................................. 59 NEW INSIGHTS AND FUTURE PERSPECTIVES ................................................ 63 Lymphatics ........................................................................................................ 63 Hemangiogenesis .............................................................................................. 65 SVENSK SAMMANFATTNING ................................................................................. 69 Bakgrund .......................................................................................................... 69 Syfte, Material och Metoder ............................................................................. 70 Resultat och Diskussion .................................................................................... 72 ACKNOWLEDGEMENTS .......................................................................................... 75 REFERENCES .............................................................................................................. 81 AIMS OF THE STUDY: ............................................................................................... 37