C-type natriuretic peptide (CNP) is an endogenous vascular modulator. In addition to vasodilation, CNP exerts multifunctions including anti-thrombus and anti-proliferation actions against vascular smooth muscle cells and myofibroblasts. Therefore, CNP is a potential therapeutic agent for the prevention of restenosis following angioplasty. The current study investigated whether local delivery of CNP, even at microgram levels about three orders of magnitude lower than doses (high milligram levels) used for systemic administration in the previous study, attenuates neointimal hyperplasia. The rabbit iliac artery was denuded, and then CNP (100 microg, n = 5) or control vehicle (n = 5) was administered locally over 20 min, via a local drug delivery catheter. During drug delivery, blood pressure was monitored with a high-fidelity micromanometer catheter. There was no significant decrease in arterial pressure immediately after the CNP administration. Four weeks after the treatment, computer-assisted morphometric analysis revealed significant reduction in the intimal area (CNP 0.44 +/- 0.27 versus control 0.96 +/- 0.20 mm2, p < 0.01), but no changes in the medial area (CNP 0.93 +/- 0.23 versus control 0.79 +/- 0.29 mm2, p = NS). This resulted in a significant decrease in the ratio of the intimal area to the medial area in CNP-treated vessels compared with control vessels (CNP 0.45 +/- 0.26 versus control 1.40 +/- 0.66, p < 0.05). Local delivery of a single low dose of CNP effectively inhibits neointimal hyperplasia with a minimal likelihood of compromising hemodynamics. Considering its multipotent actions and its role as an important regulator of the vascular system, this treatment may have a therapeutic advantage for clinical use.