THE CHICK CHORIOALLANTOIC MEMBRANE ASSAY IN THE ASSESSMENT OF ANGIOGENIC FACTORS

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Abstract
The chick chorioallantoic membrane (CAM) assay was applied for the determination of angiogenic substances. The pseudo-peroxidase activity of erythrocytes was used as a marker of blood capillaries growing in CAM during 6-14 of incubation. The CAM reaction induced by eye contact lenses, agarose gels, egg shell membranes, and Millipore and glass fiber filters implanted during days 6-10, 6-14, or 10-14 of incubation, was examined by light microscopy. Contact lenses, agarose gels and Millipore filters were soaked in [3H]histamine. The amount of radioactivity was determined in the supernatant of each carrier material incubated in vitro for 4 days, or in the CAM implants during days 10-14 of incubation. Formation of the capillary network in the chorionic epithelium was not complete on day 10 of incubation, but was complete on day 11. Contact lenses always caused capillary involution in the chorionic epithelium. Different carrier materials caused various types of inflammation, i.e. erosive, ulcerous or the granulomatous and ulcerous. In general, the carriers elicited a stronger inflammation when implanted during days 10-14 than during 6-10 or 6-14 of incubation. [3H]Histamine was released continuously from carrier materials during 4 days of incubation in vitro. In contrast, [3H]histamine release from implants in vivo lasted only for 1 day. We conclude that data obtained by the CAM assay are subjects to certain restrictions.