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Factors Influencing Blood Supply in Wound Granuloma Quantitated by a New in Vivo Technique

Department of Pharmacology, Royal College of Surgeons of England, 35–43 Lincoln's Inn Fields, London WC2A 3PN [V. M., G. P. L.], and Biology of Metastasis Laboratory, Room 536, Imperial Cancer Research Fund, P. O. Box 123, Lincoln's Inn Fields, London WC2A 3PX [V. M., I. R. H.], United Kingdom

A new quantitative assay for measuring angiogenesis in a s.c. located sponge implant in rats is described. Using this model, which detects neovascularization by measuring alterations in ¹³³Xe clearance, it has been shown that the known angiogenic factors, transforming growth factor and tumor necrosis factor , cause maximum vascularization of the sponge to occur by Day 11 postimplantation compared with Days 15 to 17 in control animals. The monokine interleukin 1 is shown to be strongly angiogenic, suggesting that more than one macrophage-derived cytokine may be the active mediator in macrophage-induced angiogenesis. Extracellular matrix proteins appear to play a role in regulating the angiogenic response such that presoaking sponges in laminin (40 µg/ml) or fibrinogen (500 µg/ml) solutions induced a significant reduction in the time taken to achieve maximum ¹³³Xe clearance values; no such enhancement of neovascularization was observed when sponges were presoaked in type IV collagen (100 µg/ml) solution. The assay described here, which is reproducible, objective, and quantitative, should be of considerable use in elucidating the molecular basis of angiogenesis regulation.

¹ To whom requests for reprints should be addressed.

Received 6/ 7/88. Revised 9/30/88. Accepted 10/11/88.

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