Julia A O'Hara

Electron paramagnetic resonance (EPR) oximetry is a promising, relatively non-invasive method of monitoring tissue partial pressure of oxygen (PtO(2)) that has proven useful in following changes in PtO(2) under various physiologic and pathophysiologic conditions. Optimal utilization of the method will be facilitated by systematic comparisons with other available methods. Here, we report on the absolute values and changes of rat brain PtO(2) using EPR oximetry and the OxyLite, an oxygen monitor based on fluorescence quenching, at adjacent locations in the same brain. EPR oximetry utilizes an implanted oxygen-sensitive material and reports tissue PtO(2) at the surface of the material. OxyLite measures PtO(2) using the fluorescence lifetime of a chromophore fixed to the tip of an optical fiber that is inserted into tissue. Measurements were made at a depth of 2-3 mm into the cortex during normoxia and during breathing of carbogen (95% O(2):5% CO(2)) followed by a return to normoxia. We conclude that in this study (1) PtO(2) values reported by the two methods are similar but not exactly the same, (2) both methods can record a baseline and rapid changes in PtO(2), (3) changes in PtO(2) induced by increasing FiO(2) from 0.26 to 0.95 (carbogen) were similar by the two methods and (4) in some rats breathing carbogen, absolute values of PtO(2) were above the sensitive range of the OxyLite method.

EPR oximetry is a promising, relatively non-invasive method for monitoring the partial pressure of oxygen in tissue (PtO2) that has proved useful in following changes under various physiologic and pathophysiologic conditions. Optimal utilization of the method will be facilitated by systematic comparisons with other available methods. Here we report on the absolute values of rat brain PtO2 using EPR and the more widely used Eppendorf polarographic microelectrode system in the same brain. EPR used an L-band (1.2 GHz) spectrometer and implanted lithium phthalocyanine (LiPc) as the oxygen-sensitive paramagnetic material. Eppendorf measurements were made by a needle probe moved vertically through the cortex at 0.5 mm intervals in three tracks including one adjacent to the location of the LiPc. Several conclusions were drawn, including, (1) the average PtO2 measured by the two methods was similar but EPR reported a significantly higher average PtO2, (2) there was poor correlation between the values in the same animal on the same side of the brain, (3) the Eppendorf reported a larger range of values and (4) the heterogeneity of oxygen levels in the brain and the areas sampled by the two methods provide an adequate explanation for the observed differences.

Photodynamic therapy (PDT) with verteporfin (lipid form of benzoporphyrin derivative,benzoporphyrin derivative monoacid ring A) was used to treat radiation-induced fibrosarcoma tumors before X-ray treatment. When verteporfin was injected 3 h before light irradiation, the tumor partial pressure of oxygen (pO(2)) rose from a pretreatment value of 2.8 +/- 1 to 15.2 +/- 6.9 mm Hg immediately after light application was complete (P = 0.048). When the optical irradiation was given 15 min after verteporfin injection, the tumor pO(2) decreased slightly after treatment [i.e., 6.8 +/- 1.6 mm Hg (pretreatment) versus 4.1 +/- 0.3 mm Hg (posttreatment)], whereas control tumor pO(2) did not change significantly. In vitro study of the cellular oxygen consumption rate before and after PDT treatment indicated that the consumption rate decreased linearly with delivered optical dose and quantitatively matched the loss of cell viability as measured by a mitochondrial tetrazolium assay. Doppler measurements show that red cell flux is still patent immediately after treatment, indicating that oxygen should still be delivered to the tumor. Computational simulations of the oxygen supply from the vessels and the consumption from mitochondrial activity confirmed that if oxygen consumption is decreased in the presence of unhindered blood flow, the tumor oxygenation should rise, and the hypoxic fraction of the tumor should decrease. Combination treatments with PDT delivered (100 J/cm(2) optical dose, with 1 mg/kg benzoporphyrin derivative monoacid ring A injected 3 h before treatment) after radiation treatment (10 Gy from 300 keV source) were compared with PDT delivered simultaneously with radiation. Tumor regrowth assay showed that the delays to reach double the tumor volume for PDT alone and radiation alone were 2.7 +/- 1.6 and 3.2 +/- 1.7 days, respectively. When radiation was given before PDT, the delay was 5.4 +/- 1.4 days, and when PDT was given at the same time as radiation, the delay was 8.1 +/- 1.5 days. This observation indicates that the combined effect in the latter case was greater than additive (P = 0.049).