Abstract

Blood substitutes are being developed using molecular solutions of modified free hemoglobin; however, anaphylactic reactions, severe renal toxicity, and hypertension have been reported in experimental models and human beings. Hypertension remains as an obstacle to the clinical use of most blood substitutes. Several investigators suggest that this effect is due to the interaction between nitric oxide and hemoglobin into the endothelial cells; hence, prevention of hemoglobin extravasation would avoid vasoconstriction. The forms of hemoglobin likely to prevent extravasation include polymerized and encapsulated Hb. Another alternative and significantly less expensive approach is the hydroxyethyl starch Hb-polymer. The aim of the present study was to compare the effect of hydroxyethyl-starch-hemoglobin with that of stroma-free hemoglobin on the in vitro contractile activity of aortic rings isolated from adult male rats. The hemoglobin-based oxygen carrier was made using stroma-free hemoglobin prepared from outdated human red cells and conjugated with 10% hydroxyethyl starch 200-260 MW. The experiments were made in thoracic segments of the aortic rings incubated with hemoglobin, starch-hemoglobin or Ringer Krebs-Bicarbonate solution (RKB) during 30 min. Smooth muscle contraction with phenylephrine and subsequent inhibition of contraction with carbachol were performed before and after incubation with hemoglobin, starch-hemoglobin, or vehicle. Incubation with hemoglobin and starch-hemoglobin significantly increased the contractile response to phenylephrine of aortic rings compared with RKB solution. The maximal response to carbachol was significantly decreased in the aortic rings incubated with either hemoglobin or starch-hemoglobin in comparison with the RKB-incubated tissues. There were no differences between the aortic rings incubated with either hemoglobin, or starch-hemoglobin. These results show that there are no differences between the effects of stroma-free hemoglobin and starch-hemoglobin on the in vitro contractile activity of aortic rings isolated from adult male rats. Our findings do not support the hypothesis that an increase in the size of the hemoglobin molecule prevents hemoglobin extravasation, and the consequent vasoconstriction due to the scavenging of nitric oxide by stroma free hemoglobin in the cellular space between endothelium and smooth muscle.

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