Inhibition by doxycycline of angiogenesis in the chicken chorioallantoic membrane (CAM)

Doxycycline, a tetracycline derivative, has many properties in addition to its antibiotic activity, including inhibition of matrix metalloproteinases (MMPs) and the ability to chelate divalent cations including Ca2+. It has been shown to inhibit endothelial cell growth in vitro, and reduce the development of experimental tumours, especially bone metastasis in a model of breast cancer. We examined the effects of doxycycline on angiogenesis in the chicken chorioallantoic membrane (CAM) model, and showed that doxycycline will cause loss of the chorionic plexus in CAMs when applied at day 8 of incubation, and the duration of this inhibition was dose-dependent. Repeated doses prolonged the inhibition, but following removal of the doxycycline there was rapid recovery of the chorionic plexus. The effects of doxycycline are in part mimicked by the MMP inhibitor 1,10-phenanthroline, and more closely by the Ca2+-chelating agent EGTA. Doxycycline was equally effective in causing loss of the chorionic plexus by day 11 in CAMs, a time at which the blood vessels are established. Doxycycline has important potential as an antiangiogenic treatment. It is capable of inhibiting angiogenesis in an in vivo model, including the removal of comparatively mature endothelial cells. The response is sensitive to the dosing regimen and the effect is rapidly reversible.