Non-invasive measurement of tumor oxygenation using embedded microparticulate EPR spin probe

We have developed a novel procedure for in situ monitoring of oxygen concentration in growing tumors by electron paramagnetic resonance (EPR)-based oximetry using embedded paramagnetic particulates. The new approach uses spin probes that are permanently embedded or implanted in the tumor. A particular advantage of this procedure is that it is non-invasive, both in terms of implantation of the probe as well as readouts of oxygen. We implanted a mixture of RIF-1 tumor cells and microparticulates of lithium phthalocyanine (LiPc) in the upper hind leg of C3H mice to grow as solid tumor. This enabled repeated measurements of oxygen concentration from the implanted site (tumor) for more than two weeks during the progression of the tumor. The particulates that were embedded in the tumor were stable and non-toxic to tumor cells. There was no apparent inhibitory effect to cell proliferation or tumor growth rate. The measurements indicated that the PO2 of the tumor decreased exponentially with tumor growth (size) and reached hypoxia (< 4 mm Hg). EPR imaging was used to identify the distribution of the particles in the tumor. The data showed a heterogeneous distribution of the probe particles within the tumor volume. Imaging of oxygen in the growing tumor demonstrated the development of significant hypoxia in the tumor within 4-6 days after inoculation. In summary, the EPR spectroscopy and imaging using embedded spin probe enabled accurate and repeated measurements of PO2 under non-perturbing conditions in growing tumors.