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Beatrice Bourghardt Peebo, Per Fagerholm, Catharina Traneus Röckert, Neil Lagali; Time-lapse In-vivo Characterization of Angiogenic Vessel Formation and Natural Vessel Regression, in an Inflammatory Corneal Model. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6432.
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To characterize angiogenic vessel formation and regression, at the single vessel level, in vivo, in an inflammatory corneal model.
Angiogenesis was induced by suture placement in one cornea of male Wistar rats under general anaesthesia. Animals were divided in two groups, to study progression and regression respectively. The progression group was monitored in vivo hourly to daily, for up to 7 days. In the regression group, sutures were removed at day 7 and animals were followed for up to three weeks. At each time point, animals were anaesthetized and the region between the limbus and suture was examined with in vivo confocal microscopy. The same corneas were removed, whole-mounted, and stained with markers for neutrophils, monocytes, macrophages, vascular endothelium and pericytes. Stained corneas were compared with morphology observed in vivo. Quantitative analysis of inflammatory cells and blood vessel lumen diameter was performed.
Within an hour after suture placement, extravasated myeloid cells patterned the stroma and soon formed endothelium-free tunnels. Four to five days later, perfused sprouts formed from parent vessels were observed, and their development into circulated vascular loops was followed. After suture removal, inflammatory cell presence significantly decreased and lumen diameter of vessels was reduced. In addition, removal of the inflammatory stimulus appeared to promote two morphologically-distinct forms of intussusception. Both in vivo and ex vivo, vessel regression and degradation were observed with mature macrophages in close proximity to vessel walls.
Following corneal hemangiogensis and regression in vivo has yielded new information about cellular level activity that is not possible to obtain with low magnification microscopy or by ex vivo analysis. We have observed stromal tunnel formation, angiogenic loop formation and natural blood vessel regression, for the first time to our knowledge, at the cellular level in vivo.
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