A New Anti‐inflammatory Test, Utilizing the Chorio‐allantoic Membrane of the Chick Embryo

Abstract

Existing methods for the detection of anti-inflammatory activity involve the use of experimental animals. These methods depend on the production of an experimental inflammation in the animal and the reduction of this inflammation by repeated doses of an anti-inflammatory agent. Such methods in common use include " the formalin foot test" (Selye, 1949), "the cotton pellet test" (Meier, Schuler & Desaulles, 1950), "the B.C.G. test " (Long & Miles, 1950), "the ultra-violet erythema test " (Wilhelmi & Domenjoz, 1951), " the granuloma pouch test " (Selye, 1953), " the pleural exudate test " (Holtkamp, Wang & Doggett, 1958), and various modifications of these methods (Buttle, D'Arcy, Howard & Kellett, 1957;Bush & Alexander, 1960;Tanaka, Kobayashi & Miyake, 1960;Newbould, 1963;and Meli, Smith & Wolff, 1964). In addition, various capillary permeability tests utilizing the spread of dyes through tissue have been introduced (Bacharach, Chance & Middleton, 1940; Seifter, Baeder & Begany, 1949) and also tests involving the anaphylactic reaction (Smith & Humphrey, 1949;Ungar, Damgaard & Hummel, 1952).The present study describes an inflammatory response induced on the chorio-allantoic membrane of the eight-day-old chick embryo, and its reduction by known anti-inflammatory agents, both steroidal and non-steroidal. These agents, all of which were effective in this test, included betamethasone, hydrocortisone, phenylbutazone, chloroquine, indomethacin and sodium salicylate. METHODSA localized inflammatory reaction was induced on the chorio-allantoic membrane of the chick embryo by the implantation of a sterile filter paper disc, followed by re-incubation in situ for 4 days. EggsFertile eggs of a cross strain were used (Sudanese bantam-English White Leghorn). These were incubated at 36-37°; a moist atmosphere was maintained by placing dishes of water on alternate shelves in the incubator; the eggs were turned twice daily. Since the ambient temperature was very high throughout the year, it was possible that incubation commenced even before the eggs were collected. For this reason, eggs were always placed in the laboratory incubator on the day that they were laid. This is contrary to general practice in more temperate climates where eggs are often stored for several days before incubation is commenced. In preliminary tests, the initial incubation period varied between 8 and 12 days.

Antiangiogenic Activity and Pharmacogenomics of Medicinal Plants from Traditional Korean Medicine

“…Plant extracts were tested for their antiangiogenic effects on quail eggs as previously described for chicken eggs with modifications [28, 29]. Briefly, the explanted embryo was placed in an incubator for 2 h at 38°C to acclimatize it to the new ambience.…”

Cytotoxicity, anti-angiogenic, apoptotic effects and transcript profiling of a naturally occurring naphthyl butenone, guieranone A

“…The embryos were cultured according to the method described by Wittmann et al [9]. GA and captopril (as positive control drug) were tested for their anti-angiogenic effects at 20 μg/ml, using chicken chorioallantoic membrane assay (CAM assay) method as described by D’Arcy and Howard [10], with modifications according to Marchesan et al [11]. The percentage inhibition of vascularization was calculated as previously described [12] (Detailed anti-angiogenic tests are available in Supporting information).…”

Anticancer Activities of Six Selected Natural Compounds of Some Cameroonian Medicinal Plants

“…Compounds 1 and 2 were tested for their anti-angiogenic effects using the method of D'Arcy and Howard [25] , with modifications according to Marchesan et al [26] . Briefly, the explanted embryo was placed in an incubator for 2 h at 38°C to acclimatize to the new ambience.…”