The ability of relaxin (RLX), which is a potent microcirculatory effector in many species including the rat, to induce de novo angiogenesis in vascularized mammalian tissue was tested using the rat mesenteric-window angiogenesis assay. RLX was administered intraperitoneally on days 1-5 at doses of 0.33, 3.3 and 33 nM. Controls received the vehicle by the same route. Groups of animals were sacrificed at the end of the 1st, 2nd and 3rd weeks. Using computer-aided microscopic morphometry including image analysis, the response was quantified by sensitive, technically independent, highly reproducible methods in terms of the vascularized area (VA), a measure of microvascular spatial extension, and the microvascular length (MVL), a measure of microvascular density. The total MVL was computed from VA × MVL. The results obtained show that RLX did not cause significant changes in any of the variables tested, regardless of dose and observation time. These findings indicate that RLX is apparently unable to mediate significant de novo angiogenesis in the system used in contrast to previously tested angiogens such as basic fibroblast growth factor, vascular endothelial growth factor, isoform 165, and tumor necrosis factor-α. In previous studies, RLX has been shown to exert antitumor activity on breast cancer cells in vitro. In the search for a possible role for RLX as an anticancer agent in vivo, it is important to know that this peptide is not angiogenic, since de novo angiogenesis is known to be a prerequisite for tumor growth and metastatic sprea