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INNOVATIVE METHODOLOGY

Oxygen distribution in murine tumors: characterization using oxygen-dependent quenching of phosphorescence

¹Department of Clinical Studies, Philadelphia, School of Veterinary Medicine, ²Department of Medicine and the Abramson Cancer Center, ³Department of Biochemistry and Biophysics, ⁴Department of Radiology, School of Medicine. University of Pennsylvania, Philadelphia, Pennsylvania

Submitted 11 October 2004 ; accepted in final form 24 November 2004

In the present work, a novel method for detecting hypoxia in tumors, phosphorescence quenching, was used to evaluate tissue and tumor oxygenation. This technique is based on the concept that phosphorescence lifetime and intensity are inversely proportional to the oxygen concentration in the tissue sample. We used the phosphor Oxyphor G2 to evaluate the oxygen profiles in three murine tumor models: K1735 malignant melanoma, RENCA renal cell carcinoma, and Lewis lung carcinoma. Oxygen measurements were obtained both as histograms of oxygen distribution within the sample and as an average oxygen pressure within the tissue sampled; the latter allowing real-time oxygen monitoring. Each of the tumor types examined had a characteristic and consistent oxygen profile. K1735 tumors were all well oxygenated, with a peak oxygen pressure of 37.8 ± 5.1 Torr; RENCA tumors had intermediate oxygen pressures, with a peak oxygen pressure of 24.8 ± 17.9 Torr; and LLC tumors were all severely hypoxic, with a peak oxygen pressure of 1.8 ± 1.1 Torr. These results correlated well with measurements of tumor cell oxygenation measured by nitroimidazole (EF5) binding and were consistent with assessments of tumor blood flow by contrast enhanced ultrasound and tumor histology. The results show that phosphorescence quenching is a reliable, reproducible, and noninvasive method capable of providing real-time determination of oxygen concentrations within tumors.

oxygen measurement; Oxyphor G2; nitroimidazole; tissue oxygenation; hypoxia

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