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Neutrophil oxidative burst evaluation during acute normovolemic hemodilution: preliminary results


In recent years there has been increasing evidence that a resuscitation strategy with different fluids can have widely divergent impacts on the immune response, neutrophil activation and tissue injury. This prospective study was undertaken to determine the neutrophil oxidative burst in the swine model during an acute normovolemic hemodilution (ANH) procedure with hydroxyethyl starch.


Twelve pigs were anesthetized, instrumented and randomized into two groups: control and hemodilution (H). The control group was only anesthetized and instrumented while animals in the ANH group were submitted to acute normovolemic hemodilution to a target hematocrit of 15% with volume replacement performed with hydroxyethyl starch 130/0.4 at a 1:1 ratio. The withdrawn blood was returned to the animals 120 minutes after the end of hemodilution. Neutrophil oxidative burst was performed with blood samples collected at the femoral vein at the following time points: before ANH (baseline), after instrumentation (INST), immediately after ANH (H), 60 minutes after ANH (60H), 120 minutes after ANH (120H), 60 minutes after blood infusion (60BI) and 120 minutes after blood infusion (120BI), and determined with a flow cytometer. Spontaneous and stimulated oxidative burst activation of neutrophils were performed with dichlorofluorescein diacetate and phorbol myristate acetate. Statistical analyses were performed using one-way analysis of variance followed by a Dunnett test or t test. A P value of 0.05 was considered statistically significant.


Spontaneous oxidative burst activity in group H increased significantly from baseline (30.19 ± 4.79) to H (57.45 ± 9.86) and 60H (56.26 ± 14.64) (P < 0.01) while the control group did not present significant variation. Between groups there were significant differences at H (ANH = 57.45 ± 9.86; control = 23.18 ± 7.16; P = 0.0007), 60H (ANH = 56.26 ± 14.64; control = 34.53 ± 9.06; P = 0.0225), 120H (ANH = 43.59 ± 5.46; control = 28.65 ± 10.44; P = 0.0220) and 60BI (ANH = 38.60 ± 1.85; control = 25.59 ± 8.12; P = 0.0082).


ANH with hydroxyethyl starch influences oxidative burst activity under experimental conditions.

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Kahvegian, M., Fantoni, T.D., Otsuki, D. et al. Neutrophil oxidative burst evaluation during acute normovolemic hemodilution: preliminary results. Crit Care 11 (Suppl 2), P40 (2007).

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