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Am J Physiol Heart Circ Physiol 283: H1292-H1301, 2002. First published May 30, 2002; doi:10.1152/ajpheart.00221.2002
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Vol. 283, Issue 4, H1292-H1301, October 2002

Comparison of effects of two hemoglobin-based O2 carriers on intestinal integrity and microvascular leakage

Ann L. Baldwin1, Elizabeth B. Wiley1, and Abdu I. Alayash2

1 Department of Physiology, College of Medicine, University of Arizona, Tucson, Arizona 85724-5051; 2 Laboratory of Plasma Derivatives, Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892

Two "blood substitutes," a diaspirin cross-linked human hemoglobin [bis(3,5 dibromosalicyl)fumarate, DBBF-Hb] and a bovine polymerized hemoglobin (PolyHbBv), advanced to clinical trials, are used in this study. Previously, we have shown that injection of DBBF-Hb into the rat circulation produces venular leakage and intestinal epithelial disruption. The purpose of this study was to determine whether PolyHbBv, currently approved for veterinary use in the United States, shows similar effects. In anesthetized Sprague-Dawley rats, the mesenteric microvasculature was perfused with DBBF-Hb (n = 6), PolyHbBv (n = 5), cyanomet Hb (CNmet-DBBF-Hb), or HEPES-buffered saline with 0.5% bovine serum albumin (HBS-BSA) (controls, n = 7) for 10 min, followed by FITC-albumin for 3 min, and then fixed for microscopy. For DBBF-Hb, the mean leak number per micrometer venule length [2.41 ± 0.33 (±SE) × 10-3] was significantly greater than for PolyHbBv (0.53 ± 0.14 × 10-3), CNmet-DBBF-Hb (0.36 ± 0.14 × 10-3), and HBS-BSA (0.12 ± 0.08 × 10-3) (P < 0.01). Corresponding quantities for leak area were 0.10 ± 0.03, 0.010 ± 0.003, 0.005 ± 0.003, and 0.02 ± 0.02 µm2/µm. In rats injected with DBBF-Hb (n = 8), intestinal epithelial integrity was significantly compromised compared with those injected with PolyHbBv (n = 5) or saline (n = 6). These results indicate that intravascular PolyHbBv produces significantly less disruption of the intestinal exchange barrier than does DBBF-Hb, probably because the heme is not so easily oxidized.

blood substitutes; fluorescence microscopy; venular leakage; electron microscopy


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