Background Purified human cross-linked hemoglobin, which is now being used in clinical trials, increases mean arterial pressure through binding of nitric oxide (NO). We postulated that binding of NO by cross-linked hemoglobin (ααHb) could also increase platelet deposition at sites of subintimal injury.

Methods and Results Male Sprague-Dawley rats were infused with ααHb (0.88 g/kg, n=8) or with the NO synthase inhibitor N^G-monomethyl-l-arginine (L-NMMA, 30 mg/kg, n=7) before undergoing microsurgical carotid endarterectomy. ¹¹¹In-labeled platelets were infused after endarterectomy, and platelet deposition was measured 20 minutes later. In control endarterectomized rats (n=8), mean platelet deposition was 7.7±0.7×10⁶/mm². Platelet deposition was significantly increased above controls in rats that received ααHb (13.2±0.9×10⁶/mm², P=.0004) and in rats infused with L-NMMA (13.9±1.0×10⁶/mm², P=.0002). The increase was prevented by infusion of l-arginine (150 mg/kg) immediately after ααHb or L-NMMA. To determine whether aspirin (ASA) blocked the increased deposition induced by ααHb, rats received oral ASA (10 mg/kg) 18 hours before endarterectomy. Platelet deposition in animals receiving ASA alone was 6.4±0.9×10⁶/mm² (n=8). This was significantly increased to 10.8±0.8×10⁶/mm² (P=.002) for the ASA-treated group that received ααHb at the time of endarterectomy (n=8). The prolonged bleeding times induced by ASA were unaffected by the infusion of ααHb.

Conclusions These data suggest that in a rat endarterectomy model, ααHb increases platelet deposition at sites of subintimal injury by binding NO. Increased deposition induced by ααHb can be prevented by administration of l-arginine but not by pretreatment with aspirin.