Differential effects of vascular growth factors on arterial and venous angiogenesis

Objective: Angiogenesis, the development of new blood vessels, has become an area of increased interest for both scientific and clinical application purposes. Proangiogenic agents, such as vascular endothelial growth factor (VEGF) and naltrexone, have been shown to effectively induce new blood vessel growth. Other growth factors, such as the endogenous opioid growth factor (OGF; [Met(5)]-enkephalin) and retinoic acid, are inhibitors of angiogenesis. The differential effects on veins and arteries, however, by any vascular growth factor, have not previously been investigated. Methods. The chick chorioallantoic membrane (CAM) assay was used for the in vivo quantitation of angiogenesis. After 3 days of incubation, fertilized chick embryos were explanted, and a 3.2-mm methylcellulose disk containing either the known angiogenic stimulators VEGF (0.2 mug, 1.0 mug) or naltrexone (0.1 mug, 5.0 mug), or the angiogenic inhibitors OGF (1.0 mug, 5.0 mug) or retinoic acid (1.0 mug) was placed onto the CAM surface. An equal volume of distilled water served as a control. After 2 days of growth, the CAM arteries and veins were identified, and images were obtained with a digital camera. Quantitative analysis of angiogenesis was performed on a 100-mm(2) area surrounding the applied disk, and the number and length of the veins and arteries were measured. Results: The angiogenic stimulators VEGF and naltrexone markedly increased both the total number and length of all blood vessels as compared with control values. The mean length of blood vessels decreased, suggesting the induction of new vessel growth. VEGF and naltrexone proportionately increased vein and arterial angiogenesis, maintaining artery/vein ratios for vessel number and length that were unchanged compared with controls. The angiogenic inhibitors, OGF and retinoic acid, notably decreased the total number and length of blood vessels in the CAM preparations. However, these compounds had a disproportionately greater inhibitory effect on arterial angiogenesis as reflected in decreased artery/vein ratios for vessel number and length. Conclusions: The angiogenic stimulators VEGF and naltrexone induce development of veins and arteries in a proportional manner. In contrast, the angiogenic inhibitors OGF and retinoic acid demonstrated a greater inhibitory effect on arterial as compared with venous angiogenesis. Such differential effects on angiogenesis may be important in both defining mechanisms of action and designing therapeutic interventions.