The chick Chorioallantoic Membrane (CAM) is a double layer of

mesoderm formed by the fusion of the allantois and the chorion of the developing
embryo. This double layer is enriched with vascular supply, which connects to the
embryonic circulation via the allantoic arteries and veins (Ribatti, 2001). Due to
this property, CAM is frequently used for in vivo study of angiogenesis.

An effect on angiogenesis is indicated by a change in vascular density in

and around the CAM as a result of the addition of test substances applied to it
(Irvin, 2015, p.7). In terms of anti-angiogenesis, a positive result is indicated by
vascular occlusion (Nowak-Slowinska, 2015, p. 11), while pro-angiogenic effect
is measured by growth of the vessels and sprouting ability (Manjunathan, 2015, p.
4).

Numerous studies about anti-angiogenic properties of different substances

utilized the CAM assay. One of the substances assessed for its anti-angiogenic
potential is the PSP-2
(1,2bis(bis(dimethylphosphorothioylmethyl)phosphino)ethane), a copper chelator
by observing that it suppressed the branching of the vessels (Heuberger, 2019,
p.8).

Crenolanib, a tyrosine kinase inhibitor, showed anti-angiogenic effects by

decreasing the average sprout length (Berndsen, 2019).

Another study observed the ability of opioids in inhibiting angiogenesis by

observing the decrease of capillary density after administration in different
concentrations (Karaman, 2017).

Strong anti-angiogenic activity is also exhibited by bioactive compounds

of different species of plants using CAM assay, among those is Ginger (Zinciber
officinale) which exhibited its anti-angiogenic property by decreasing the CAM
region and blood vessel diameter. The decreased CAM region is proportional the
angiogenesis activity (Bashir, 2017).

The CAM assay offers many advantages, the most prominent of which is
the accessibility and rapid growth, as the CAM develops in a relatively short
period of time, making it suitable for studies involving developmental pathways
(Nowak-Sliwinska, 2015, p.3). This also adds to the fact that chicken eggs are
inexpensive, making the method suitable for large scale screening (Stryker, 2019,
p. 9). The CAM lacks innervation, making the embryo insensitive to pain. This
offers an advantage because ethical issues does not pose as a limitation (Kunz,
2019, p.2).