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Automated evaluation of angiogenic effects mediated by VEGF and PlGF homo- and heterodimers.

The effects of growth factors on the blood vessel pattern of chick chorioallantoic membrane (CAM) were assessed with a fast and automated method (extended counting method, XCM; Sandau, 1996) that measures complexity, without assumptions about a fractal structure. XCM is a reliable measure of complexity not only in theory but also in practice: (1) it is robust with respect to thresholding; (2) it shows reduced variance due to pattern translation and rotation; (3) its properties come close to requirements of fractal geometry. It hence is superior to established fractal methods for distinguishing effects induced by various isoforms of vascular endothelial growth factor (VEGF121 and VEGF165), placenta growth factor (PlGF) isoforms, and control treatment. We here show that VEGF homo- and heterodimers and VEGF121/PlGF1 heterodimers increase vascular complexity, whereas PlGF1 and PlGF2 are not effective. PlGF1 and VEGF121 did not mutually influence each other when applied in adjacent fields on the same CAM. Since blood vessels in the CAM originate via nonfractal growth processes, their growth should be analyzed accordingly.