Experimental studies have implicated a functional role for the extracellular matrix glycoprotein thrombospondin-1 (TSP-1) in vascular smooth muscle cell proliferation and migration. We therefore sought to determine if TSP-1 might represent a specific component of the fibroproliferative tissue typically associated with restenosis lesions from human coronary and peripheral arteries. Positive immunostaining for TSP-1 was limited to hypocellular plaques typical of primary atherosclerosis; in contrast, such staining was nearly absent from the loose extracellular matrix of the fibroproliferative tissue typical of restenotic lesions. Only a small fraction of vascular smooth muscle cells in either primary or restenotic lesions demonstrated a cellular staining pattern for TSP-1, which was also observed in control studies performed in cell culture and in atherosclerotic rabbit arteries examined 3 days after experimental balloon angioplasty. Double-staining for TSP-1 and proliferating cell nuclear antigen in studies of human beings disclosed that only a small portion of proliferating cell nuclear antigen-positive cells also stained for TSP-1. The observations made in this series of specimens thus indicate that TSP-1 is not a major component of the extracellular matrix of human restenotic tissues, even when such specimens demonstrate evidence of hypercellularity or ongoing cellular proliferation. Because most restenosis specimens, however, were retrieved ≥1 month after the primary intervention, a functional role for TSP-1 in smooth muscle cell proliferation or migration at the early stages of lesion development is still possible. (Am Heart J 1998;135:357-64.)