Abstract-Thrombospondin-1 (TSP1) limits the angiogenic and vasodilator activities of NO. This activity of TSP1 can be beneficial in some disease states, but endogenous TSP1 limits recovery of tissue perfusion following fixed ischemic injury in dorsal skin flaps in mice. Using mice lacking the TSP1 receptors CD36 or CD47, we now show that CD47 is the necessary receptor for limiting NO-mediated vascular smooth muscle relaxation and tissue survival following ischemic injury in skin flaps and hindlimbs. We further show that blocking CD47 or TSP1 using monoclonal antibodies and decreasing CD47 expression using an antisense morpholino oligonucleotide are effective therapeutic approaches to dramatically increase survival of soft tissue subjected to fixed ischemia. These treatments facilitate rapid vascular remodeling to restore tissue perfusion and increase skin and muscle viability. Thus, limiting CD47-dependent antagonism of NO-mediated vasodilation and vascular remodeling is a promising therapeutic modality to preserve tissues subject to ischemic stress. (Circ Res. 2007;100:712-720.)Key Words: nitric oxide Ⅲ thrombospondin-1 Ⅲ ischemic tissue survival Ⅲ CD47 Ⅲ therapeutics T issue viability requires continuous perfusion, which in turn depends on vascular tone, sufficient intravascular volume, and adequate blood oxygenation. 1-3 The contractile status of arterial smooth muscle is the major determinant of vascular tone, with venous tone playing a lesser role. 4,5 Underperfusion of soft tissues is the leading cause of tissue necrosis and secondary delayed wound healing in surgical patients. 6 The complications incurred can be substantial and life threatening. 7 Complications of inadequate tissue perfusion are multiplied in the elderly and patients with hypertension and diabetes because of the general vasculopathies associated with these disease processes. 8,9 Current therapies to improve vascular perfusion combine surgical vessel manipulation/bypass with vasodilators that relax vascular smooth muscle cells (VSMCs). 10,11 The bioactive gas NO is a potent vasodilator 12 that activates soluble guanylate cyclase. The increased cGMP activates cGMPdependent protein kinases and thereby decreases VSMC sensitivity to intracellular Ca 2ϩ , leading to relaxation of contractile proteins. [13][14][15][16] We recently reported that NO/cGMP signaling in VSMCs and endothelial cells is potently inhibited by the secreted protein thrombospondin-1 (TSP1). [17][18][19] We further showed that endogenous TSP1 limits the ability of NO to increase skeletal muscle perfusion and blood oxygen levels in vivo. 20 Following surgically induced acute ischemia in random dorsal skin flaps, endogenous TSP1 also limits tissue survival and recovery of tissue oxygenation. Ischemic tissue survival could be improved by increasing NO levels using isosorbide dinitrate, but the degree of tissue necrosis in treated wild-type mice remained higher than in TSP1-null mice, which achieved essentially complete flap survival following this treatment.To further improve s...