Time-Lapse In Vivo Imaging of Corneal Angiogenesis: The Role of Inflammatory Cells in Capillary Sprouting

Peebo, Beatrice Bourghardt; Traneus-Röckert, Catharina; Lagali, Neil

doi:10.1167/iovs.10-6101

Cited by 32 publications

(47 citation statements)

References 44 publications

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“…G. At day 12, a similar appearance as in day 7 was observed. (Scale bar = 50 µm) and budding was observed, followed by lumen relaxation and sprouting at budding sites, as previously reported [33]. In the two days following suture removal and bevacizumab treatment, myeloid cells infiltrated the stroma, lumens re-expanded, and blood flow increased.…”

Section: Fig 12supporting

confidence: 79%

“…In a separate experiment, subjconjunctival injection of saline into two healthy rat eyes prompted myeloid cell infiltration into the cornea. The same effect was observed in Fig 12 in the days following injection, and resulted in re-expansion of lumens and stronger blood flow, paralleling the inflammation observed after initial suture placement [33] . Presumably, the broader anti-inflammatory activity of triamcinolone may have aided regression in this group following this second inflammatory insult, compared to a possible lack of VEGFA-specific inhibition by bevacizumab in the other group.…”

Section: In Vivo Imaging Of Human Corneal Capillary Regression: Reporsupporting

confidence: 75%

“…Digital images were recorded at 5 frames/s, and the probed region was adjusted in lateral, transverse, and axial directions during image capture to locate and follow the path of vessels from the suture area to the limbus. IVCM has been shown to be a valuable tool for imaging of various cells in transparent tissue, noninvasively, in a time-lapse manner, without the need for cell labeling [33,34]. With IVCM it is possible to identify and follow the same structures and thereby observe dynamic processes in live tissue.…”

Section: In Vivo Confocal Microscopymentioning

confidence: 99%

“…We have recently developed a model of inflammatory angiogenesis in the rat cornea and a technique whereby single angiogenic vessels are studied in a time lapse manner [33]. The in vivo model preserves the complexity of the physiologic milieu which includes tissue proteins, inflammatory cells, mesenchymal cells, pre-existing vessels and the circulation, all of which may contribute factors regulating the growth and regression of vessels.…”

Section: Introductionmentioning

confidence: 99%

“…In this study, we applied our model and technique to characterize the morphology of regression in vivo, in a manner allowing vessels and cell-vessel interactions to be observed over time in the same tissue. Specifically, we sought to i) examine temporal changes in the morphology of vessels in natural and therapeutically-induced regression, ii) track the presence of inflammatory, myeloid-lineage cells [33] during regression, and iii) examine the post-regression microstructure of the affected tissue. In our model, we abruptly terminated angiogenesis in a strongly pro-angiogenic inflammatory environment, relatively early during the course of inflammation.…”

Section: Introductionmentioning

confidence: 99%

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Cellular level characterization of capillary regression in inflammatory angiogenesis using an in vivo corneal model

2011

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Abstract:In this study, we introduce a technique for repeated, microscopic observation of single regressing capillaries in vivo in inflamed murine corneas. Natural capillary regression was initiated by removal of inflammatory stimulus during an active pro-angiogenic phase, while the additional impact of anti-angiogenic treatment with triamcinolone or bevazicumab was investigated. Capillaries regressed naturally within one week and treatments did not further enhance the natural regression. Morphologically, early-phase regression was characterized by significant lumen narrowing and a significant reduction in CD11b+ myeloid cell infiltration of the extracellular matrix. By one week, vascular remodeling occurred concomitant with CD11b+CD68+KiM2R+ mature macrophage localization on capillary walls. Empty conduits without blood flow, positive for collagen IV and devoid of vascular endothelium and pericytes, were apparent in vivo and by three weeks were more numerous than perfused capillaries. By three weeks, macrophages aggregated around remaining perfused capillaries and were observed in apposition with degrading capillary segments. Abrupt termination of capillary sprouting in our regression model further revealed vascular endothelial abandonment of sprout tips and perfused capillary loop formation within a single angiogenic sprout, possibly as an intussusceptive response to cessation of the stimulus. Finally, we observed lumen constriction and macrophage localization on capillary walls in vivo in a clinical case of corneal capillary regression that paralleled findings in our murine model.

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Section: Fig 12supporting

confidence: 79%

Section: In Vivo Imaging Of Human Corneal Capillary Regression: Reporsupporting

confidence: 75%

Section: In Vivo Confocal Microscopymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cellular level characterization of capillary regression in inflammatory angiogenesis using an in vivo corneal model

2011

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Umbilical Cord Blood-Derived Mononuclear Cells Exhibit Pericyte-Like Phenotype and Support Network Formation of Endothelial Progenitor Cells In Vitro

et al. 2015

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Umbilical cord blood represents a promising cell source for pro-angiogenic therapies. The present study examined the potential of mononuclear cells (MNCs) from umbilical cord blood to support endothelial progenitor cell (EPC) microvessel formation. MNCs were isolated from the cord blood of 20 separate donors and selected for further characterization based upon their proliferation potential and morphological resemblance to human vascular pericytes (HVPs). MNCs were screened for their ability to support EPC network formation using an in vitro assay (Matrigel™) as well as a reductionist, coculture system consisting of no additional angiogenic cytokines beyond those present in serum. In less than 15% of the isolations, we identified a population of highly proliferative MNCs that phenotypically resembled HVPs as assessed by expression of PDGFR-β, NG2, α-SMA, and ephrin-B2. Within a Matrigel™ system, MNCs demonstrated pericyte-like function through colocalization to EPC networks and similar effects as HVPs upon total EPC tubule length (p = 0.95) and number of branch points (p = 0.93). In a reductionist coculture system, MNCs served as pro-angiogenic mural cells by supporting EPC network formation to a significantly greater extent than HVP cocultures, by day 14 of coculture, as evidenced through EPC total tubule length (p <0.0001) and number of branch points (p < 0.0001). Our findings are significant as we demonstrate mural cell progenitors can be isolated from umbilical cord blood and develop culture conditions to support their use in microvascular tissue engineering applications.

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Endothelial cells expressing low levels of CD143 (ACE) exhibit enhanced sprouting and potency in relieving tissue ischemia

2014

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The sprouting of endothelial cells from pre-existing blood vessels represents a critical event in the angiogenesis cascade. However only a fraction of cultured or transplanted endothelial cells form new vessels. Moreover it is unclear if this results from a stochastic process or instead relates to certain endothelial cells having a greater angiogenic potential. This study investigated whether there exists a sub-population of cultured endothelial cells with enhanced angiogenic potency in vitro and in vivo. First, endothelial cells that participated in sprouting, and non-sprouting cells, were separately isolated from a 3D fibrin gel sprouting assay. Interestingly, the sprouting cells, when placed back into the same assay, displayed a 7-fold increase in the number of sprouts, as compared with control cells. Angiotensin converting enzyme (CD143) was significantly down regulated on sprouting cells, as compared to regular endothelial cells. A subset of endothelial cells with low CD143 expression was then prospectively isolated from an endothelial cell culture. Finally, these cells were found to have greater potency in alleviating local ischemia, and restoring regional blood perfusion when transplanted into ischemic hindlimbs, as compared with unsorted endothelial cells. In summary, this study indicates that low expression of CD143 can be used as a biomarker to identify an endothelial cell subpopulation that is more capable to drive neovascularization.

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Time-Lapse In Vivo Imaging of Corneal Angiogenesis: The Role of Inflammatory Cells in Capillary Sprouting

Abstract: Time-lapse in vivo corneal confocal microscopy can be used to track a temporal sequence of events in corneal angiogenesis. The technique has revealed potential roles for myeloid cells in promoting vessel sprouting in an inflammatory corneal setting.

Cited by 32 publications

References 44 publications

Cellular level characterization of capillary regression in inflammatory angiogenesis using an in vivo corneal model

Cellular level characterization of capillary regression in inflammatory angiogenesis using an in vivo corneal model

Umbilical Cord Blood-Derived Mononuclear Cells Exhibit Pericyte-Like Phenotype and Support Network Formation of Endothelial Progenitor Cells In Vitro

Endothelial cells expressing low levels of CD143 (ACE) exhibit enhanced sprouting and potency in relieving tissue ischemia

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