Journal List > J Korean Orthop Assoc > v.31(2) > 1105185

Park, Lee, Song, Lee, and Chun: Hydrogen Peroxide Production in Neutrophils after Tourniquet Release

Abstract

The use of lower extremity tourniquets for procedures of the lower leg is considered routine in orthopedic surgery, but. lower extremity tourniquets do harm occasionally. While the tourniquet is inflated, metabolic changes such as increased PaCO2, lactic acid, and serum potassium and decreased level of PaO2 and pH occur in the ischemic limb. Deflation of tourniquet results in release of anaerobic metabolic products during ischemia into systemic circulation. In this ischemia/reperfusion situation, oxygen free radicals could potentially be produced during the reperfusion period by several mechanisms. One of these mechanisms is release of intracellular superoxide or hydrogen peroxide by activated neutrophils in the area. These reactive oxygen species(ROS) could be a causative factor for the postreperfusion no-flow, lung injury, induction of tourniquet shock, etc. The purpose of this clinical study was to investigate the effect of tourniquet deflation on the hemodynamic changes, changes of blood gas analysis, and hydrogen peroxide production using flow cytometric analysis of fluorescent DCF(Dichlorofluorescein). Quantitative analysis of fluorescent DCF was performed in resting and fMLP(N-formyl-methyonyl-leucyl-phenylalanine) or PMA(phorbol myristate acetate) stimulated neutrophils. Also differences of these factors between two groups of tourniquet time, one is less than one hour and the other more than one to two hours, were analysed. The hemodynamics (blood pressure, pulse rate), arterial P02, bicarbonate, base excess, and hydrogen peroxide production showed no significant change before and after tourniquet release(p>0.05). Arterial pH and PaCO2 decreased significantly until 10 and 5 minutes after tourniquet release, respectively(p<0.05).
Tourniquet time didn’t reveal any significances differences. These results indicate that tourniquet application with 400mmHg pressure and less than 2 hours does not release significant hydrogen peroxide into systemic circulation during reperfusion period after tourniquet release.

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Fig. 1
Time course of increasing fluorescent DCF in resting and stimulated PMNL(poly morphonuclear leukocyte), this figure shows there is no difference of hydrogen per oxide production expressed as fluorescent DCF before and after tourniquetrelease. fMLP(10-8 M/L):N-formyl-methyonyl-luecyl-phenylala nine PMA(300 ng/ml/): Phorbol myritate acetate
jkoa-31-388f1.tif
Table 2
Changes of Vital Signs Before and After Tourniquet Release
BTR 1 min 3 min 5 min 10 min 15 min 30 min ATR
BP(S) 124±4 113±4 114±3 114±3 122±4 120±3 122±5
BP(D) 74±3 67±2 67±2 67±2 72±3 76±3 75±3
PR 78±4 79±3 79±4 78±4 80±4 79±4 77±3

Values are expressed as Mean ± SE

BP(D):Diastolic Blood Pressure(mmHg)

ATR:After tourniquet release

BP(S):Systolic Blood Pressure(mrnHg)

PR:Pulse Rate(beat/min) BTR:Before tourniquet release

Table 3
Changes of ABGA Before and After Tourniquet Release(FiO2=0.5)
BTR 1 min 3 min 5 min 10 min 15 min 30 min ATR
pH 7.37 ±.01 7.32 ±.01* 7.30 ±01* 731 ±.01* 7.32±.01* 7.34±.02 7.35 ±.02
PCO2 35.7 ±2.8 41.9±4.8* 42.4±3.3* 42.0±5.5* 40.0± 4.5 37.8±3.4 36.2 ±2.4
PO2 233±8 217± 9 232 ± 7 228 ± 9 230± 11 253 ± 10 247± 15
HCO3 18.8 ±0.9 19.1 ±0.91 18.8±0.9 18.3 ±0.9 18.2±0.9 18.3 ±1.2 18.8± 1.0
BE -1.2±1.5 -1.5 ± 2.0 -1.4±1.1 -1.4±2.0 -1.6±1.9 -2,1 ±2.4 -1.9± 1.3

Values are expressed as Mean ± SE

BTR:Before tourniquet release

ABGA: Arterial blood gas analysis

ATR: After tourniquet release

BE:Base excess

* P<0.05 compared with control value

Table 4
Time Course of Increasing Fluorescent DCF in Resting and Stimulated Neutrophil
Time BTR 1 min 3 min 5 min 10 min 15 min 30 min ATR
Rest 70.95 ±3.73 69.26±3.23 69.25±3.26 67.62 ±3.68 68.98 ± 3.10 66.02 ±3.10 64.37±2.86
fMLP 71.78±3.76 67.44±3.05 70.03±3.59 71.85±4.00 6915±3.32 67.90±3.07 67.37±3.65
PMA 380.61 ±35.91 364.46 ±34.19 357.62±27.82 347.78±29.49 347.77 ±29.49 344.73 ±24.98 334.17±24.04

Values are expressed as Mean ± SE

PMA(Phorbol Myritate Acetate):300 ng/ml

fMLP(N-formyl-Methyonyl-luecyl-phenylalanine): 10-s M/L

BTR:Before tourniquet release ATR: After tourniquet release

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