Supplementary Material for: Vascular Patterning as Integrative Readout of Complex Molecular and Physiological Signaling by VESsel GENeration Analysis

The molecular signaling cascades that regulate angiogene?sis and microvascular remodeling are fundamental to nor?mal development, healthy physiology, and pathologies such as inflammation and cancer. Yet quantifying such complex, fractally branching vascular patterns remains difficult. We re?view application of NASA’s globally available, freely down?loadable VESsel GENeration (VESGEN) Analysis software to numerous examples of 2D vascular trees, networks, and tree-network composites. Upon input of a binary vascular image, automated output includes informative vascular maps and quantification of parameters such as tortuosity, fractal dimension, vessel diameter, area, length, number, and branch point. Previous research has demonstrated that cytokines and therapeutics such as vascular endothelial growth factor, basic fibroblast growth factor (fibroblast growth factor-2), transforming growth factor-beta-1, and steroid triamcinolone acetonide specify unique “fingerprint” or “biomarker” vascular patterns that integrate dominant signaling with physiological response. In vivo experimental examples described here include vascular response to kera?tinocyte growth factor, a novel vessel tortuosity factor; an?giogenic inhibition in humanized tumor xenografts by the anti-angiogenesis drug leronlimab; intestinal vascular in?flammation with probiotic protection by Saccharomyces boulardii, and a workflow programming of vascular architec?ture for 3D bioprinting of regenerative tissues from 2D im?ages. Microvascular remodeling in the human retina is de?scribed for astronaut risks in microgravity, vessel tortuosity in diabetic retinopathy, and venous occlusive disease.