Patterns of angiogenic response to mast cell granule constituents

Agents derived from mast cell granule constituents, and compound 48/80 which stimulates release of mast cell granules, have been used by us to develop new methods for quantitating angiogenesis in the chick chorioallantoic membrane. Two of these methods provide different insights, demonstrating different patterns of response to dosage and over time, produced by different agents. Counting mesenchymal blood vessels is convenient for obtaining dose-response data. Histamine and compound 48/80 have been shown previously to give a sigmoid dose-response curve resulting in a plateau before the lethal dose. This contrasts with the effect of porcine sodium heparin (Evans Biologicals) which results in a minor increase then a relative decline in vessel number due to a failure of growth. Here, the ability to produce angiogenesis or antiangiogenesis appears to be dose-dependent. Measurement of the changes in DNA synthesis, leading to visible angiogenesis, may be performed once the optimal angiogenic dose is known, and again distinctive patterns of response with different agents have been found. Histamine results in a fall then rise to a peak at 36 hr. We now show that two types of heparin each produce a peak at 12 hr. Compound 48/80 results in a distinctive pattern that looks like a composite of the histamine and especially the heparin effects, and this suggests that both are relevant to induction of angiogenesis by mast cells. The elicitation of this pattern of response also provides a method, additional to electron microscopy, for discovering whether or not an angiogenic substance is likely to operate via mast cell stimulation. Such characteristic patterns offer a new way of classifying angiogenic substances.