Background: The PEGylated cell-free carboxyhemoglobin gas exchanger SanguinateTM (SG) has multiple beneficial actions that may improve stroke outcome including anti-inflammatory, vasodilatory and oxygen delivery capacity. We investigated the ability of SG to improve infarction in spontaneously hypertensive rats (SHR) that are known to have vasoconstricted pial collaterals and poor stroke outcome.
Methods: SHR were treated IV with SG (8 ml/kg; n=8) or vehicle (Lactated Ringer’s; n=6) plus tPA (0.9 mg/ml) after 30 min of proximal middle cerebral artery occlusion (MCAO) by filament occlusion. Pial collateral and reperfusion flow were measured for 2 hrs of occlusion and 2 hrs reperfusion using dual laser Doppler probes in the core (Bregma -2mm, lateral +4mm) and collateral (Bregma +2mm, lateral +3mm) territories. Collateral openings (number and duration (min)) were assessed off cerebral blood flow (CBF) tracings, defined as independent of changes in blood pressure (Fig. 1; arrows show openings). Infarction was measured using triphenyltetrazolium chloride (TTC). Animals were anesthetized with chloral hydrate and ventilated to maintain blood gases within normal ranges.
Results: Vehicle-treated SHR had incomplete reperfusion and poor collaterals that was improved by SG. During MCAO, SG increased the number of collateral openings from 1.0±0.8 to 3.1±0.8 (p=0.03) and duration of openings from 6.0±5.8 to 35.4±9.8 min (p=0.04; Fig. 2). Reperfusion CBF was -43±6% of baseline in vehicle-treated, but only -7±19% of baseline in SG-treated animals. Improved collateral and reperfusion flow with SG was associated with decreased infarct volume vs. vehicle (28.8±3.2% vs. 18.8±2.3%; p<0.05). The presence of tPA had no effect in either group.
Conclusions: Smaller infarction with early SG treatment may be related to its ability to open constricted collaterals in SHR and improve reperfusion. Thus, SG may be able to extend the time window for endovascular or tPA treatment.