Quantitative intracellular oxygen availability before and after 5-aminolevulinic acid skin photodynamic therapy.

BACKGROUND: During photodynamic therapy (PDT) oxygen is transformed into reactive oxygen species (ROS) to induce cellular apoptosis in (pre)malignant cells. Real time oxygen availability measurement is clinically available with the Cellular Oxygen Metabolism (COMET) monitor. METHODS: Primary objective is to show that mitochondrial oxygen availability (mitoPO2) measurement is possible during clinical ALA-PDT. The secondary aim was to determine the pain sensation, because it is the most commonly reported side effect of PDT. Before and after the two fraction PDT treatment, with a 2-hour dark period, mitoPO2 was measured and reported pain was documented with a visual analog scale (VAS) 0-100. RESULTS: Nine patients were included. Before the first PDT session the median signal quality was [IQR] 55.0% [34.2-68.0], which decreased after session one to 0% [0.0-10.0]. MitoPO2 was 40.0 [17.7-53.8] mmHg and increased afterwards to 61.8 [38.2-64.8] mmHg. This likely the result of the delay time between the illumination stop and the mitoPO2 measurements in a vasodilated, visibly red lesion. Before session two signal quality was 10.4% [0-20.15], 40% lower than at the start. In 5 patients the signal quality after session 2 was too low because of photobleaching and insufficient regeneration of PpIX, median 0% [0-10]. Subjects reported low median VAS scores, all below 3, directly after the mitoPO2 measurements. CONCLUSION: With COMET we were able to reliably measure mitochondrial oxygen concentrations during photodynamic therapy. Signal quality drastically decreases after a PDT session because of PpIX deterioration during the illumination phase.

Mitochondrial oxygen monitoring with COMET: verification of calibration in man and comparison with vascular occlusion tests in healthy volunteers.

Mitochondria are the primary consumers of oxygen and therefore an important location for oxygen availability and consumption measurement. A technique has been developed for mitochondrial oxygen tension (mitoPO2) measurement, incorporated in the COMET. In contrast to most textbooks, relatively high average mitoPO2 values have been reported. The first aim of this study was to verify the validity of the COMET calibration for mitoPO2 measurements in human skin. The second aim was to compare the dynamics of mitoPO2 to several other techniques assessing tissue oxygenation. Firstly, we performed a two-point calibration. Mitochondrial oxygen depletion was achieved with vascular occlusion. A high mitoPO2 was reached by local application of cyanide. MitoPO2 was compared to the arterial oxygen partial pressure (PaO2). Secondly, for deoxygenation kinetics we compared COMET variables with the LEA O2C, SenTec OxiVenT™ and Medtronic INVOS™ parameters during a vascular occlusion test. 20 healthy volunteers were recruited and resulted in 18 datasets (2 times 9 subjects). The lowest measured mitoPO2 value per subject had a median [IQR] of 3.0 [1.0-4.0] mmHg, n = 9. After cyanide application the mitoPO2 was 94.1 mmHg [87.2-110.9] and did not differ significantly (n = 9, p = 0.5) from the PaO2 of 101.0 [98.0-106.0] mmHg. In contrast to O2C, OxiVenT™ and INVOS parameters, mitoPO2 declined within seconds with pressure on the probe. The kinetics from this decline are used to mitochondrial oxygen consumption (mitoVO2). This study validates the calibration of the COMET device in humans. For mitoVO2 measurements not only blood flow cessation but application of local pressure is of great importance to clear the measurement site of oxygen-carrying erythrocytes.

The effect of storage time of human red cells on intestinal microcirculatory oxygenation in a rat isovolemic exchange model.

OBJECTIVE: To determine whether the storage time of human leukodepleted red blood cell concentrates compromises intestinal microvascular oxygen concentration oxygen (muPo(2)) during isovolemic exchange transfusion at low hematocrit. DESIGN: Prospective, randomized, controlled study. SETTING: University research institute laboratory. SUBJECTS: Male Wistar rats. INTERVENTIONS: Intestinal muPo(2) was determined by Pd-porphyrin phosphorescence life-time measurements. MEASUREMENTS AND MAIN RESULTS: Rats were brought near to a state of oxygen supply dependency by hemodilution with a pasteurized plasma protein solution to a hematocrit of 14.3 +/- 1.1% (n = 24). Subsequently, an isovolemic exchange transfusion with human leukodepleted red blood cells, stored for 2-6 days (fresh, n = 8), 2-3 wks (intermediate, n = 8), or 5-6 wks (old, n = 8), was performed to determine whether intestinal muPo(2) would be preserved. Immunologic reactions were avoided by washing the red blood cell concentrates three times before use. Isovolemic exchange with fresh and intermediate red blood cells maintained muPo(2) whereas old cells decreased muPo(2) with 26%. Subsequent transfusion with red blood cells (hematocrit approximately 60%) until reaching a hematocrit of 32.4 +/- 2.1 % (n = 24) increased intestinal muPo(2) in all three groups to the same extent between 28% and 32%. No changes in red blood cell deformability, as determined by a Laser-assisted Optical Rotational Cell Analyzer, could be demonstrated during 5 wks of storage. CONCLUSION: This study shows that at low hematocrit, the oxygen-delivering capacity of human red blood cells stored 5-6 wks is reduced compared with fresh cells and red blood cells stored for an intermediate period. Although red blood cells stored for 2-3 wks are completely devoid of 2,3-diphosphoglycerate, their oxygen-delivering capacity to the intestines was the same as fresh red blood cells. Our study showed that red blood cell deformability was preserved during storage, suggesting that other mechanisms may account for the observed decrease in oxygen delivery by red blood cells stored 2-3 wks.