HET-CAM bioassay as in vitro alternative to the croton oil test for investigating steroidal and non-steroidal compounds.
In this study the irritation phenomena at the chorioallantoic membrane of incubated hen's eggs as an in vitro model (HET-CAM assay) were investigated in comparison to the in vivo croton oil test by including hydrocortisone, indomethacin, phenylbutazone, acetylsalicylic acid, rutin, quercetin, apigenin, and p-coumaric acid as steroidal and non-steroidal test substances. For the first time the two methods were compared in a valid way with the perspective of a realistic reduction of animal experiments. It should be investigated whether an in vitro-in vivo correlation exists and, if there is any possibility, to replace the in vivo model by an in vitro test system. Both bioassays were able to demonstrate the anti-inflammatory potency of the constituents tested. The determination of the anti-inflammatory activity of all compounds in the two test systems showed individual trends of inhibitory effects. However, the in vitro HET-CAM test was much more sensitive in comparison to the in vivo croton oil test. The croton oil test gave dose-effect correlations in the anti-inflammatory substances investigated. The modified HET-CAM assay did not provide clear dose-effect ratios. The HET-CAM assay is an inexpensive test being easy to manage after a short practical training. Because of its sensitivity the HET-CAM assay could be considered a suitable tool for qualitative testing of the anti-inflammatory activity of substances if no appropriate dose-effect curves are required. From these results it can be concluded that the different courses of the dose-effect curves may be primarily due to different mechanisms of action.
Brantner
Adelheid H
AH
Institute of Pharmacognosy, University of Graz, A-Graz, Trieste. adelheid.brantner@kfunigraz.ac.at
Quehenberger
Franz
F
Chakraborty
Asima
A
Polligger
Jutta
J
Sosa
Silvio
S
Della Loggia
Roberto
R
D000482:Allantois / Q000187:drug effects; D000826:Animal Testing Alternatives; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D001681:Biological Assay / Q000379:methods*; D002642:Chick Embryo; D002823:Chorion / Q000187:drug effects; D003436:Croton Oil; D007249:Inflammation; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013256:Steroids / Q000032:analysis*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D013256:Steroids; D003436:Croton Oil